Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mediator complex subunit 1 promotes oral squamous cell carcinoma progression by activating MMP9 transcription and suppressing CD8 + T cell antitumor immunity

doi: 10.1186/s13046-024-03191-9

Figure Lengend Snippet: MED1 is upregulated in OSCC patients and associated with poor prognosis. A Analysis of the mRNA levels of MED1 (cancer vs. normal) in multiple solid cancers from Oncomine Database. B Analysis of the MED1 expression level (tumor vs. normal) in multiple solid cancers from TIMER Database. C mRNA expression of MED1 in 3 samples (GSE30784, GSE25099, and GSE10121) from the GEO database. D Immunohistochemical staining of MED1 in OSCC tissue microarray (TMA). E Quantification of immunostaining results. F Kaplan–Meier analysis of overall survival of OSCC patients from GEPIA database stratified by MED1 levels. G Kaplan–Meier analysis of overall survival of OSCC patients from UALCAN database stratified by MED1 levels. H Kaplan–Meier analysis of overall survival of OSCC patients from TCGA database stratified by MED1 levels. Scale bar = 200 µm (10 ×), and 50 µm (40 ×), Bars = means ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001. OSCC, oral squamous cell carcinoma

Article Snippet: Human OSCC cell lines Cal-27, UPCI-SCC-090, SCC-9, UPCI-SCC-154 and mouse OSCC cell line SCC-7 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Microarray, Immunostaining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mediator complex subunit 1 promotes oral squamous cell carcinoma progression by activating MMP9 transcription and suppressing CD8 + T cell antitumor immunity

doi: 10.1186/s13046-024-03191-9

Figure Lengend Snippet: MED1 is highly expressed in human OSCC cell line and has no significant effect on metastatic OSCC cells proliferation. A Cell morphology of human oral keratinocytes (HOK), nonmetastatic oral squamous cell carcinoma UPCI-SCC-090, and metastatic oral squamous cell carcinoma UPCI-SCC-154. B mRNA levels of MED1 in HOK and four OSCC lines tested by qRT-PCR. n = 3 independent experiments. C Protein levels of MED1 in HOK and four OSCC lines tested by WB. D CCK8 to detect SCC-9 cell proliferation change after MED1 knockdown. n = 3 independent experiments. E CCK8 to detect UPCI-SCC-154 cell proliferation change after MED1 knockdown. n = 3 independent experiments. F Plate colony formation assay to detection of clonality in SCC-9 cells following MED1 knockdown. n = 3 independent experiments. G Plate colony formation assay to detection of clonality in UPCI-SCC-154 cells following MED1 knockdown. n = 3 independent experiments. H Immunofluorescence staining and quantitative analysis of proliferation marker Ki67 after MED1 knockdown in SCC-9 cells. n = 3 independent experiments. I Immunofluorescence staining and quantitative analysis of proliferation marker Ki67 after MED1 knockdown in UPCI-SCC-154 cells. n = 3 independent experiments. Scale bar = 100 µm (10 ×), and 50 µm (40 ×). Bars = means ± SD. ns means nonsignificant, *** P < 0.001, **** P < 0.0001

Article Snippet: Human OSCC cell lines Cal-27, UPCI-SCC-090, SCC-9, UPCI-SCC-154 and mouse OSCC cell line SCC-7 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Quantitative RT-PCR, Knockdown, Colony Assay, Immunofluorescence, Staining, Marker

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mediator complex subunit 1 promotes oral squamous cell carcinoma progression by activating MMP9 transcription and suppressing CD8 + T cell antitumor immunity

doi: 10.1186/s13046-024-03191-9

Figure Lengend Snippet: MED1 knockdown inhibits metastatic OSCC cells migration and invasion in vitro. A Cell scratch assay to examine SCC-9 cells migration ability after MED1 knockdown. B Quantitative analysis of the cell migration ratio in ( A ). n = 3 independent experiments. C Cell scratch assay to examine UPCI-SCC-154 cells migration ability after MED1 knockdown. D Quantitative analysis of the cell migration ratio in ( C ). n = 3 independent experiments. E Transwell invasion assay to examine SCC-9 cells invasion ability after MED1 knockdown. F Quantitative analysis of the number of invasive cells in (E). n = 3 independent experiments. G Transwell invasion assay to examine UPCI-SCC-154 cells invasion ability after MED1 knockdown. H Quantitative analysis of the number of invasive cells in ( G ). n = 3 independent experiments. I MMP2 and MMP9 gene expression assessed by qRT-PCR in SCC-9 cells after MED1 knockdown. n = 3 independent experiments. J MMP2 and MMP9 gene expression assessed by qRT-PCR in UPCI-SCC-154 cells after MED1 knockdown. n = 3 independent experiments.Scale bar = 200 µm (10 ×). Bars = means ± SD.* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human OSCC cell lines Cal-27, UPCI-SCC-090, SCC-9, UPCI-SCC-154 and mouse OSCC cell line SCC-7 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Knockdown, Migration, In Vitro, Wound Healing Assay, Transwell Invasion Assay, Gene Expression, Quantitative RT-PCR

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mediator complex subunit 1 promotes oral squamous cell carcinoma progression by activating MMP9 transcription and suppressing CD8 + T cell antitumor immunity

doi: 10.1186/s13046-024-03191-9

Figure Lengend Snippet: MED1 regulates metastatic OSCC cells migration and invasion through the modulation of MMP9. A MMP9 gene expression assessed by qRT-PCR. n = 3 independent experiments. B MMP9 protein expression assessed by WB. n = 3 independent experiments. C The luciferase activity of MMP9 promoter detected by luciferase reporter assay. n = 3 independent experiments. D The recruitments of cofactors AP-1(c-Jun/c-Fos) on MMP9 promoter in SCC-9 cells and UPCI-SCC-154 cells by CHIP assay and qRT-PCR. n = 3 independent experiments. E Cell scratch assay to determine the changes in migration abilities of SCC-9 cells after adding exogenous MMP9 and quantitative analysis. n = 3 independent experiments. F Cell scratch assay to determine the changes in migration abilities of UPCI-SCC-154 cells after adding exogenous MMP9 and quantitative analysis. n = 3 independent experiments. G Transwell assay to determine the changes in invasion abilities of SCC-9 cells after adding exogenous MMP9 and quantitative analysis ( H ). n = 3 independent experiments. I Transwell assay to determine the changes in invasion abilities of UPCI-SCC-154 cells after adding exogenous MMP9 and quantitative analysis ( J ). n = 3 independent experiments. Scale bar = 200 µm (10 ×). Bars = means ± SD.* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human OSCC cell lines Cal-27, UPCI-SCC-090, SCC-9, UPCI-SCC-154 and mouse OSCC cell line SCC-7 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Migration, Gene Expression, Quantitative RT-PCR, Expressing, Luciferase, Activity Assay, Reporter Assay, Wound Healing Assay, Transwell Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mediator complex subunit 1 promotes oral squamous cell carcinoma progression by activating MMP9 transcription and suppressing CD8 + T cell antitumor immunity

doi: 10.1186/s13046-024-03191-9

Figure Lengend Snippet: Graphic abstract of molecular mechanisms of MED1 promoting tumor progression in OSCC. A For untreated metastatic OSCC cells, MED1 facilitates MMP9 expression by promoting the transcription of MMP9, resulting in heightened migration and invasion of metastatic OSCC cells, just like strengthen the destructive force of "bandits". Additionally, MED1 upregulates PD-L1 expression via activation of the Notch signaling pathway, resulting in diminished cytotoxicity of CD8 + T cells within the tumor microenvironment and consequent attenuation of anti-tumor immunity responses. This behavior is similar to weaken the fighting capacity of "police". B For MED1 knockdown metastatic OSCC cells, the transcription of MMP9 is inhibited, leading to decreased migration and invasion of metastatic OSCC cells, just like weaken the destructive force of "bandits". This inhibition also leads to a decrease in PD-L1 expression through the suppression of Notch signaling pathway, indirectly enhancing the cytotoxic activity of CD8 + T cells in the TME and strengthening antitumor immunity responses. This behavior is similar to strengthen the fighting capacity of "police". MMP9: matrix metallopeptidase 9, POL II: RNA polymerase II, TATA: TATA box, PD-L1: programmed cell death ligand 1, NICD: Notch intracellular domain, PD-1: programmed cell death 1, IFN-γ: interferon-γ, IL-2: Interleukin-2

Article Snippet: Human OSCC cell lines Cal-27, UPCI-SCC-090, SCC-9, UPCI-SCC-154 and mouse OSCC cell line SCC-7 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Migration, Activation Assay, Knockdown, Inhibition, Activity Assay

Journal: Hereditas

Article Title: LPAR2 correlated with different prognosis and immune cell infiltration in head and neck squamous cell carcinoma and kidney renal clear cell carcinoma

doi: 10.1186/s41065-022-00229-w

Figure Lengend Snippet: Information of the Selected GEO Datasets

Article Snippet: GSE30784 , Chen C, et al. (2011) , OSCC , Affymetrix Human Genome U133 Plus 2.0 Array , 167/62.

Techniques: Microarray, Expressing

Journal: Current Issues in Molecular Biology

Article Title: Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C 44 Mab-108 for Immunohistochemistry

doi: 10.3390/cimb45030121

Figure Lengend Snippet: Immunohistochemical analysis using C 44 Mab-108 and C 44 Mab-46 against oral squamous cell carcinoma (OSCC) tissues. After antigen retrieval, the sections were incubated with 10 µg/mL of C 44 Mab-108 ( A , B , I , J ), 1 µg/mL of C 44 Mab-46 ( C , D , K , L ), and without the primary antibody (control) ( E , F , M , N ) followed by treatment with the Envision+ kit. The color was developed using 3,3′-diaminobenzidine tetrahydrochloride (DAB), and the sections were counterstained with hematoxylin. ( G , H , O , P ) Hematoxylin and eosin (HE) staining. ( Q , R ) Blocking of the C 44 Mab-108 reactivity to OSCC tissue by the CD44 peptide (aa 271–290) containing the C 44 Mab-108 epitope. After antigen retrieval, sections were incubated with C 44 Mab-108 (10 μg/mL) or C 44 Mab-108 (10 μg/mL) plus human CD44 peptide (aa 271–290, 10 μg/mL) followed by treatment with the Envision+ kit. The color was developed using DAB, and sections were counterstained with hematoxylin. Scale bar = 100 µm.

Article Snippet: The formalin-fixed paraffin-embedded (FFPE) OSCC tissue microarray (Product Code: OR601c, US Biomax Inc., Rockville, MD, USA) and the esophageal tissue microarray (Product Code: BC02011, US Biomax Inc.) were deparaffinized in xylene (Sigma-Aldrich Corp.) and rehydrated.

Techniques: Immunohistochemical staining, Incubation, Staining, Blocking Assay

Journal: Cancer

Article Title: Sirtuin-3 (SIRT3), a Novel Potential Therapeutic Target for Oral Cancer

doi: 10.1002/cncr.25676

Figure Lengend Snippet: Sirtuin-3 (SIRT3) is overexpressed in oral squamous cell carcinoma (OSCC). (A) Immunoblots reveal the levels of sirtuins (SIRT1-7) in the OSCC cell lines HSC-3, UM-SCC-1, and UM-SCC-17B and in normal human oral keratinocytes (K). β-Actin served as loading control. (B) These representative samples show ( Top ) SIRT3 expression levels in OSCC (tongue) and ( Bottom ) in normal tissues. Immunoglobulin G (IgG) served as a negative control. Scale bars = 200 μm for low-magnification photomicrographs (left column); 50 μm for high-magnification photomicrographs (middle and right columns). (C) These charts illustrate the percentage of normal and OSCC tissue specimens that expressed SIRT3 ( Top ) in all samples and ( Bottom ) in tongue samples, as determined by immunohistochemical staining. Note that SIRT3 staining intensities were designated as low (L) or high (H) from the data provided in .

Article Snippet: Immunohistochemical analyses were performed to determine the expression of SIRT3 and SIRT7 in human normal and OSCC tissues using OSCC tissue microarrays (OR601 and HN241; US Biomax, Inc., Rockville, Md) and the Histostatin Kit (95-6143; Zymed Laboratories, South San Francisco, Calif) according to the manufacturer's instructions.

Techniques: Western Blot, Expressing, Negative Control, Immunohistochemical staining, Staining

Journal: Cancer

Article Title: Sirtuin-3 (SIRT3), a Novel Potential Therapeutic Target for Oral Cancer

doi: 10.1002/cncr.25676

Figure Lengend Snippet: The sirtuin inhibitors sirtinol and nicotinamide (NAM) inhibit cell growth and proliferation and induce apoptosis. (A) Shown are ( Top ) phase-contrast images and ( Bottom ) histograms that reveal the morphology of cell growth and cell proliferation (%) of the HSC-3, UM-SCC-1, and UM-SCC-17B oral squamous cell carcinoma (OSCC) cell lines after treatment with sirtuin inhibitors sirtinol (50 μM) or NAM (40 mM) for 16 hours and 24 hours, respectively (original magnification, ×100). (B) Cell death-detection ELISA assays were used to measure DNA fragmentation in cells after treatment with ( Top ) sirtinol and ( Bottom ) NAM as indicated. ** P ≤ .01; *** P ≤ .001; OD, optical density.

Article Snippet: Immunohistochemical analyses were performed to determine the expression of SIRT3 and SIRT7 in human normal and OSCC tissues using OSCC tissue microarrays (OR601 and HN241; US Biomax, Inc., Rockville, Md) and the Histostatin Kit (95-6143; Zymed Laboratories, South San Francisco, Calif) according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Cancer

Article Title: Sirtuin-3 (SIRT3), a Novel Potential Therapeutic Target for Oral Cancer

doi: 10.1002/cncr.25676

Figure Lengend Snippet: Sirtuin-3 (SIRT3) down-regulation inhibits cell growth and proliferation and promotes apoptosis in oral squamous cell carcinoma (OSCC) cells. (A) ( Top ) Immunoblots reveal the transfection efficiency of SIRT3 in the UM-SCC-1 and UM-SCC-17B OSCC cell lines 36 hours after transfection with SIRT3 small interfering RNA (siSIRT3) or nontargeting control (siControl) (150 nM). β-Actin served as loading control. ( Middle ) Cells were transfected as indicated and were cultured for 1 week, then stained with crystal violet, and photographed. ( Bottom ) Numbers of colonies are presented as the percentage of colonies obtained relative to controls. (B) Cell death-detection ELISA assays were used to measure DNA fragmentation in the cells after transfection. ** P ≤ .01; *** P ≤ .001; OD, optical density.

Article Snippet: Immunohistochemical analyses were performed to determine the expression of SIRT3 and SIRT7 in human normal and OSCC tissues using OSCC tissue microarrays (OR601 and HN241; US Biomax, Inc., Rockville, Md) and the Histostatin Kit (95-6143; Zymed Laboratories, South San Francisco, Calif) according to the manufacturer's instructions.

Techniques: Western Blot, Transfection, Small Interfering RNA, Cell Culture, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cancer

Article Title: Sirtuin-3 (SIRT3), a Novel Potential Therapeutic Target for Oral Cancer

doi: 10.1002/cncr.25676

Figure Lengend Snippet: Sirtuin-3 (SIRT3) down-regulation enhances the sensitivity of oral squamous cell carcinoma (OSCC) to radiation and cisplatin-induced cytotoxicity. OSCC cells (UM-SCC-1 and UM-SCC-17B) were either untransfected or transfected with small interfering SIRT3 (siSIRT3) (150 nM) and were treated with either (A) ionizing radiation (2.5 grays) or (B) cisplatin (20 μM) for 24 hours, and cytotoxicity was determined by using the QUANT Cell Proliferation Assay Kit (Invitrogen). ** P ≤ .01; *** P ≤ .001.

Article Snippet: Immunohistochemical analyses were performed to determine the expression of SIRT3 and SIRT7 in human normal and OSCC tissues using OSCC tissue microarrays (OR601 and HN241; US Biomax, Inc., Rockville, Md) and the Histostatin Kit (95-6143; Zymed Laboratories, South San Francisco, Calif) according to the manufacturer's instructions.

Techniques: Transfection, Proliferation Assay

Journal: Cancer

Article Title: Sirtuin-3 (SIRT3), a Novel Potential Therapeutic Target for Oral Cancer

doi: 10.1002/cncr.25676

Figure Lengend Snippet: Sirtuin-3 (SIRT3) down-regulation reduces oral squamous cell carcinoma (OSCC) tumor burden in vivo. (A) These immunoblots show SIRT3 expression levels in 4 clones from UM-SCC-17B cell lines that were stably transfected with scrambled-short hairpin RNA (Scr-shRNA) or SIRT3-shRNA after 10 days of selection using Puromycin. β-Actin served as loading control. (B) Mice were injected with OSCC cells that stably expressed Scr-shRNA or SIRT3-shRNA. Images show ( Top ) the superficial growth of tumors in the head and neck region of the mice, ( Middle ) dissected tumors in situ, and ( Bottom ) dissected and isolated tumors from the 2 groups.

Article Snippet: Immunohistochemical analyses were performed to determine the expression of SIRT3 and SIRT7 in human normal and OSCC tissues using OSCC tissue microarrays (OR601 and HN241; US Biomax, Inc., Rockville, Md) and the Histostatin Kit (95-6143; Zymed Laboratories, South San Francisco, Calif) according to the manufacturer's instructions.

Techniques: In Vivo, Western Blot, Expressing, Clone Assay, Stable Transfection, Transfection, shRNA, Selection, Injection, In Situ, Isolation

Journal: Advanced Science

Article Title: Long Noncoding RNA HITTERS Protects Oral Squamous Cell Carcinoma Cells from Endoplasmic Reticulum Stress‐Induced Apoptosis via Promoting MRE11‐RAD50‐NBS1 Complex Formation

doi: 10.1002/advs.202002747

Figure Lengend Snippet: Identification of ER stress‐related lncRNAs. A) Western blot and B) qPCR confirmed that different concentrations of TM (5, 10 µg mL −1 ) or short time TM exposure could significantly induce ER stress of both SCC25 and CAL27 cells. C) Heatmap and D) volcano plot of HTA 2.0 lncRNA microarray for gene expression profiles of nontreated and TM‐treated SCC25 cells. E) qPCR confirmed the results of microarray that HITTERS was upregulated by treating cells with TM (5, 10 µg mL −1 ) for 6 h. F–J) qPCR showed F,G) treating different cells with TM (10 µg mL −1 ) for 6 h or H–J) treating SCC25 with different types of ER stress inducer for 6 h could upregulate both ER stress marker and HITTERS . The Student t ‐test was used for analyzing the difference in (F) and (G). One‐way Analysis of Variance (ANOVA) test and Dunnett t ‐test was used for (B), (E), and (H)–(J). For (E)–(G), all p < 0.001. Note: *, P < 0.05; **, P < 0.01, ***, P < 0.001.

Article Snippet: Human OSCC cell lines SCC25 and CAL27 were purchased from American Type Culture Collection.

Techniques: Western Blot, Microarray, Gene Expression, Marker

Journal: Advanced Science

Article Title: Long Noncoding RNA HITTERS Protects Oral Squamous Cell Carcinoma Cells from Endoplasmic Reticulum Stress‐Induced Apoptosis via Promoting MRE11‐RAD50‐NBS1 Complex Formation

doi: 10.1002/advs.202002747

Figure Lengend Snippet: HITTERS shares the same promoter with HERPUD1 . A) Dual‐luciferase reporter assay showed the potential promoter DNA fragments of HITTERS lacked transcription activity no matter threated with TM (10 µg mL −1 , 6 h) or not. One‐way ANOVA and Dunnett t ‐test were used, “Vector” was the control, all differences were none significant. B) qPCR results showed that time‐course change of HITTERS was identical to HERPUD1, but significant differed from BIP. Cells were treated with TM (10 µg mL −1 ) and measured every 3 h. C) qPCR results showed HITTERS and HERPUD1 had a strong coexpression pattern in 48 OSCC samples. Liner‐regression test was used. D) Dual‐luciferase reporter assay showed the promoter of HERPUD1 had strong transcription activity and responded obviously to TM (10 µg mL −1 , 6 h). Student t ‐test was used. E) qPCR results indicated that depleting HERPUD1 promoter in HEK293 by CRISPR/Cas9 system significantly suppressed the expression level of both HERPUD1 and HITTERS , no matter treated with or without TM (10 µg mL −1 , 6 h). One‐way ANOVA and Dunnett t ‐test were used, “Vector” was the control. qPCR results indicated knockdown of F) ATF4, G) ATF6, and H) XBP1s significantly suppressed HITTERS expression. Cells were transfected with siRNA for 48 h and then treated with TM (10 µg mL −1 ) for 6 h. Student t ‐test was used. I) qPCR results showed treating cells with 5aza (10 × 10 −6 m , 24 h) for inhibiting DNMTs significantly promoted HITTERS expression. Student t ‐test was used. Note: ns, no significance; **, P < 0.01, ***, P < 0.001.

Article Snippet: Human OSCC cell lines SCC25 and CAL27 were purchased from American Type Culture Collection.

Techniques: Luciferase, Reporter Assay, Activity Assay, Plasmid Preparation, Control, CRISPR, Expressing, Knockdown, Transfection

Journal: Advanced Science

Article Title: Long Noncoding RNA HITTERS Protects Oral Squamous Cell Carcinoma Cells from Endoplasmic Reticulum Stress‐Induced Apoptosis via Promoting MRE11‐RAD50‐NBS1 Complex Formation

doi: 10.1002/advs.202002747

Figure Lengend Snippet: HITTERS promotes OSCC progression and correlates with OSCC clinicopathological features. A) CCK8, B) colony formation test, and C) EdU incorporation test confirmed that knockdown of HITTERS suppressed OSCC proliferation in vitro. D,E) Stably knockdown of HITTERS significantly suppressed tumor volume and tumor weight in D) SCC25 and E) CAL27 subcutaneous xenograft model. F) Wound healing test and G) transwell assay confirmed knockdown of HITTERS suppressed OSCC migration and invasion ability in vitro. Stably knockdown of HITTERS significantly suppressed H) SCC25 and I) CAL27 pulmonary metastasis nodule formation ability in vivo. The green fluorescent protein (GFP) fluorescense imaging of lungs was also presented. J) qPCR analyzed the relative fold of HITTERS in 48 OSCC tissues normalized by their paired adjacent normal tissues. K) qPCR results indicated the expression of HITTERS in OSCC was higher than their paired adjacent normal tissues. Paired t ‐test was used. L) Kaplan–Meier curves showed high HITTERS expression had poor over‐all survival. Log‐rank test was used. For (A)–(I) Student t ‐test was used. For (A), (D), and (E), Student t ‐test was used for each time point. Note: *, P < 0.05; **, P < 0.01, ***, P < 0.001.

Article Snippet: Human OSCC cell lines SCC25 and CAL27 were purchased from American Type Culture Collection.

Techniques: Knockdown, In Vitro, Stable Transfection, Transwell Assay, Migration, In Vivo, Imaging, Expressing

Journal: Advanced Science

Article Title: Long Noncoding RNA HITTERS Protects Oral Squamous Cell Carcinoma Cells from Endoplasmic Reticulum Stress‐Induced Apoptosis via Promoting MRE11‐RAD50‐NBS1 Complex Formation

doi: 10.1002/advs.202002747

Figure Lengend Snippet: HITTERS attenuates ER stress induced apoptosis. A) Cell viability (measured by CCK8) of SCC25 and CAL27 were significantly suppressed by TM (5, 10, and 20 µg mL −1 for 24 and 48 h) in a dose‐ and time‐dependent manner. One‐way ANOVA and Dunnett t ‐test were used for each time point. B) Western blot showed the ER stress marker BIP and apoptosis marker of SCC25 and CAL27 cells were significantly upregulated by TM (10 µg mL −1 ) in a time‐dependent manner. c‐PARP, cleaved PARP. C,D) Intraperitoneally injection of TM twice a week significantly suppressed tumor volume and tumor weight in C) SCC25 and D) CAL27 subcutaneous xenograft model. E,F) Under non‐ER stress condition, HITTERS would not affect apoptosis; however, depletion of HITTERS in TM (10 µg mL −1 , 24 h) treated E) SCC25 and F) CAL27 cells significantly promoted apoptosis. Apoptosis was measured by Annexin‐V/PI double staining and flow cytometry. Proportion of R3 + R5 is considered apoptosis. Cells were transfected with siRNA for 48 h and then treated with TM (10 µg mL −1 , 24 h). Two‐way ANOVA and Sidak's multiple comparisons test were used. G) Knocking‐down HITTERS significantly suppressed cell viability and promoted apoptosis marker expression; H) whereas overexpressing HITTERS obtained the opposite effect. Cells were transfected with siRNA for 48 h and then treated with TM (10 µg mL −1 , 24 h). Cell viability differences were measured by CCK8 using Student t ‐test. Stably knockdown of HITTERS causing I) SCC25 and J) CAL27 more sensitive to ER stress in vivo, reflected by a significantly reduction in tumor volume and tumor weight in subcutaneous xenograft model. All BALB/c nude mice were intraperitoneally injected with TM, twice a week, after tumor bearing. Student t ‐test was used. Note: *, P < 0.05; **, P < 0.01, ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Human OSCC cell lines SCC25 and CAL27 were purchased from American Type Culture Collection.

Techniques: Western Blot, Marker, Injection, Double Staining, Flow Cytometry, Transfection, Expressing, Stable Transfection, Knockdown, In Vivo

Journal: Advanced Science

Article Title: Long Noncoding RNA HITTERS Protects Oral Squamous Cell Carcinoma Cells from Endoplasmic Reticulum Stress‐Induced Apoptosis via Promoting MRE11‐RAD50‐NBS1 Complex Formation

doi: 10.1002/advs.202002747

Figure Lengend Snippet: HITTERS regulates ER stress related DDR. A) qPCR and B) Western blot confirmed that knocking‐down HITTERS did not significantly change the mRNA and protein expression of HERPUD1 . For qPCR, cells were transfected with siRNA for 48 h. Student t ‐test was used for qPCR. C,D) Knocking‐down HITTERS did not significantly change the mRNA level of important UPR regulator in both C) SCC25 and D) CAL27 cells. Cells were transfected with siRNA for 48 h and then treated with TM (10 µg mL −1 ) for 6 h. E) GSEA results for RNA‐sequencing profiles. SCC25 cells were transfected with HITTERS siRNA or negative control siRNA for 48 h and then treated with TM (10 µg mL −1 ) for 12 h. F,G) 2',7'‐dichlorofluorescein staining and TUNEL assay indicated TM (10 µg mL −1 , 24 h) treatment promoted F) ROS production and G) DNA breaks. Student t ‐test was used. H) Induction of ER stress by TM (10 µg mL −1 ) significantly promoted the level of DNA damage marker γ ‐H2AX and suppressed DNA repair proteins in a time‐dependent manner. I) Knocking‐down HITTERS significantly suppressed DNA repair protein and promoted DNA damage marker expression; J) whereas overexpressing HITTERS obtained the opposite effect. Cells were transfected with siRNA or plasmid for 48 h and then treated with TM (10 µg mL −1 ) for 24 h. Note: ns, no significance; ***, P < 0.001.

Article Snippet: Human OSCC cell lines SCC25 and CAL27 were purchased from American Type Culture Collection.

Techniques: Western Blot, Expressing, Transfection, RNA Sequencing, Negative Control, Staining, TUNEL Assay, Marker, Plasmid Preparation

Journal: Advanced Science

Article Title: Long Noncoding RNA HITTERS Protects Oral Squamous Cell Carcinoma Cells from Endoplasmic Reticulum Stress‐Induced Apoptosis via Promoting MRE11‐RAD50‐NBS1 Complex Formation

doi: 10.1002/advs.202002747

Figure Lengend Snippet: HITTERS binds to and regulates the formation of MRN complex under ER stress. A) The schematic representation of modified ChIRP‐MS. B) qPCR showed the modified ChIRP method retrieved about 30–50% of HITTERS . Western blot confirmed that HITTERS binds to both MRE11 and RAD50, but not NBS1 nor glyceraldehyde‐3‐phosphate dehydrogenase. Cells were treated with TM (10 µg mL −1 , 6 h) before harvesting for ChIRP‐MS. For qPCR, Student t ‐test was used for each primer. ***, P < 0.001. C) RIP assay of MRE11 and RAD50. Western blot confirmed MRE11 and RAD50 were successfully precipitated. qPCR indicated HITTERS were significantly enriched by MRE11 and RAD50. Cells were treated with TM (10 µg mL −1 , 6 h) before harvesting for RIP. For qPCR, Student t ‐test was used. ***, P < 0.001. D,E) Co‐IP results showed the interaction between MRE11 and RAD50 in CAL27 cells were reduced after HITTERS knockdown. The interaction between MRE11 and NBS1 remained no change after HITTERS knockdown. Cells were transfected with siRNA for 48 h and then treated with TM (10 µg mL −1 ) for 6 h. H) Diagrams of full‐length HITTERS and the truncations in MS2bs‐MS2bp RNA pull‐down assay. I) The reconstructed plasmids containing 12XMS2 tag and full‐length HITTERS and truncations with the correct sizes are indicated. J,K) Immunoblot analysis for RAD50 and MRE11 in the protein samples pulled down by different HITTERS truncations. J) SCC25 and K) CAL27 cells were transfected with two plasmids for 48 h and treated with TM (10 µg mL −1 ) for 6 h.

Article Snippet: Human OSCC cell lines SCC25 and CAL27 were purchased from American Type Culture Collection.

Techniques: Modification, Western Blot, Co-Immunoprecipitation Assay, Knockdown, Transfection, Pull Down Assay

Journal: Gastroenterology Report

Article Title: Transcriptomic biomarkers for predicting response to neoadjuvant treatment in oesophageal cancer

doi: 10.1093/gastro/goaa065

Figure Lengend Snippet: Studies reporting mRNAs associated with response to neoadjuvant therapy in OAC and OSCC

Article Snippet: Two studies evaluated predictive biomarkers in OSCC using an Agilent microarray platform [ , ].

Techniques: Expressing, Microarray

Journal: Gastroenterology Report

Article Title: Transcriptomic biomarkers for predicting response to neoadjuvant treatment in oesophageal cancer

doi: 10.1093/gastro/goaa065

Figure Lengend Snippet: Studies reporting miRNAs associated with response to neoadjuvant therapy in OAC and OSCC

Article Snippet: Two studies evaluated predictive biomarkers in OSCC using an Agilent microarray platform [ , ].

Techniques: Biomarker Assay, miRNA RT, Expressing, Microarray

Journal: Biomolecules

Article Title: Nicotinamide N-Methyltransferase in Head and Neck Tumors: A Comprehensive Review

doi: 10.3390/biom11111594

Figure Lengend Snippet: Summary of the literature data on the role of NNMT in HNT.

Article Snippet: Roberg K et al., 2008 [ ] , OSCC , Genomic study , Genomic microarray [Affymetrix HG-Focus chip] (LK0412 cell line) , NNMT expression is 74-fold change higher in OSCC than in normal oral keratinocytes..

Techniques: Expressing, Microarray, Immunostaining, Chromatography, Mass Spectrometry, In Vitro, Activity Assay, In Vivo, Over Expression, Binding Assay, Mutagenesis, Migration, Gas Chromatography-Mass Spectrometry

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: A . TP63 mRNA levels were significantly upregulated in OSCC per four independent microarrays that were retrieved from Oncomine. B . Increased TP63 mRNA expression was revealed in several types of human cancers per Oncomine. Log 2 median-centered intensity represents the TP63 mRNA expression levels. C . TP63 expression in human OSCCs in cohort #1 (n=103) and noncancerous adjacent tissues (NAT, n=28). Representative immunohistochemistry images for TP63 staining in NAT, different localization in one OSCC tissue and OSCC tissues from various pathological differentiation, lymph nodes statuses and clinical stages are shown. Red arrows represent tumor budding cells. The representative images of low expression (upper panel) or high expression (lower panel) of TP63 are shown. Original magnification 400×. D-G . A vertical scatter plot is presented to demonstrate the relative expression levels of TP63 in NATs and OSCCs (D), OSCC tissues from patients with different pathological differentiation (E), lymph nodes metastasis statuses (F) and disease stages (G). H . Kaplan-Meier curves for the disease-free survival (DFS) of OSCC patients with low TP63 expression (n=45) vs. high TP63 expression (n=58). * P <0.05, ** P <0.01.

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: Correlation of TP63 expression with clinical and pathological variables of OSCC patients in cohort #1 (n=103)

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: Expressing

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: Cox regression models of patients with OSCC for clinical and pathological parameters (cohort#1, n=103)

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: Expressing

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: The impact of ΔNp63 on cell proliferation was determined using CCK-8 A, D . and colon-formation assays B, E . C, F . Transwell assays were used to evaluate the effect of ΔNp63 on cell migration or invasion (upper panel). The relative numbers of cells that migrated or invaded are shown (lower panel). The results were obtained in three independent experiments and are shown as the mean±SD. G, J . The effects of ΔNp63 on the tumor xenografts that were generated with subcutaneous injections of the SCC9 or SCC15 stable cells are shown. H, K . Growth curves of the tumor volumes, which were measured every 7 days post-injection, are shown. I, L . Tumor weights are represented. M . Representative photographs of the hepatic metastases, and hematoxylin and eosin (H&E) stains, the immune staining of human pan-CK in metastatic livers of mice (n=5 per group) in the models generated by way of tail vein injection of SCC9 stable cells (left panel). Original magnification 400×. Quantification of the number of liver metastases (right panel). * P <0.05, ** P <0.01.

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: CCK-8 Assay, Migration, Generated, Injection, Staining

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: A, B . The impact of ΔNp63 on the expression of representative stem cell markers was assessed by Western blot (left panel). Quantification of the relative protein expression levels is shown (right panel). C, D . Representative images of the tumor spheres formed by SCC9 or SCC15 stable cells lines after culturing for 2 weeks (left panel). Statistical analysis of the average diameters of the spheres (right panel). Original magnification 50×. E . Expression of the ΔNp63, Ki67, CD44, KLF4, and SOX2 proteins in the transplanted tumors was detected by IHC, and representative micrographs are presented. Original magnification 400×.

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: Expressing, Western Blot

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: A . The differential miRNA profiles of the SCC9-LvNC and SCC9-LvΔNp63 cells were characterized with a PCR-based miRNA microarray. B . Six differentially expressed miRNAs were selected from the miRNA profiles and validated by qRT-PCR. C . qRT-PCR analysis showing the miR-138-5p and ΔNp63 expression levels in a panel of 7 oral cancer cells and NOK. D . Relationship between ΔNp63 expression and miR-138-5p expression levels in a panel of 7 oral cancer cells and NOK was analyzed by Spearman order correlation. E . qRT-PCR analysis showing the miR-138-5p expression levels in the SCC9 or SCC15 stable cell lines. F . qRT-PCR analysis showing pre-miR-138 (miR-138-1 and miR-138-2) expression levels in SCC15-shΔNp63 compared with the control group (SCC15-shNC). G . TP63 binding motif sequence (upper panel). The schematic diagram represents two putative ΔNp63 binding sites, which occupy the upstream region of the miR-138-2 promoter, and the pGL3-basic plasmid, which contains the wild type (pGL3-Wt) or the mutated binding sites (pGL3-Mut-BS1, pGL3-Mut-BS2, pGL3-Mut-BS1+2) (lower panel). H . Luciferase reporter assays were performed in SCC9 cells after co-transfection with pGL3-basic reporters that contained the wild-type or the mutated binding sites and the ΔNp63 plasmid (pEZ-ΔNp63). The data were normalized to data from a simultaneously delivered Renillar luciferase expression plasmid and determined relative to an empty vector (pGL3-basic). I . ChIP assays using antibodies against ΔNp63 or the IgG control were performed in SCC15 cells. Binding of ΔNp63 to the TP21 promoter (positive control) and the two binding sites (BS1 and BS2) on miR-138-2 promoter region was confirmed by PCR using specific primers to the binding sites. The PCR gel shows the amplification of the ΔNp63 binding sites (left panel). The qPCR for ΔNp63 binding sites (right panel). All results are presented as the mean±SD obtained from three independent experiments. * P <0.05, ** P <0.01.

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: Microarray, Quantitative RT-PCR, Expressing, Stable Transfection, Control, Binding Assay, Sequencing, Plasmid Preparation, Luciferase, Cotransfection, Positive Control, Amplification

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: A, B . The effects of ΔNp63 and ΔNp63/miR-138-5p on cell proliferation were determined in SCC9 or SCC15 cells using the CCK-8 (A) and colony formation (B) assays. C . The effects of ΔNp63 and ΔNp63/miR-138-5p on the cell migration or invasion of SCC9 or SCC15 cells were measured with transwell assays. D . The impact of ΔNp63 and ΔNp63/miR-138-5p on tumor sphere formation and E . the exppression of representative stem cell markers. F . Protein expression level quantification. G, H . Analysis of cancer stem-like cells marker ALDH (G) and CD133 (H) in SCC9 and SCC15 cells. Percentage of ALDH positive cells were calculated against DEAB control. Original magnification 50×. The results are presented as the mean±SD of three independent experiments. * P <0.05, ** P <0.01.

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: CCK-8 Assay, Migration, Expressing, Marker, Control

Journal: Oncotarget

Article Title: Interplay between ΔNp63 and miR-138-5p regulates growth, metastasis and stemness of oral squamous cell carcinoma

doi: 10.18632/oncotarget.15752

Figure Lengend Snippet: A . Representative micrographs showing the expression of ΔNp63 and miR-138-5p in OSCC tissues with different pathological differentiation or different localizations from one OSCC tissue specimen. Original magnification 400×. B . The relationship between the ΔNp63 expression and miR-138-5p expression levels was analyzed using the Spearman order correlation. C . Kaplan-Meier survival curves for the DFS of OSCC patients with low TP63 expression (n=50) vs. high TP63 expression (n=60) D . Kaplan-Meier survival curves for the DFS of OSCC patients with low ΔNp63 expression (n=50), high ΔNp63 expression and high miR-138-5p expression (n=20), or high ΔNp63 expression but low-miR-138-5p expression (n=40). The survival differences were analyzed using a log-rank test. E . A schematic illustration of the interplay between ΔNp63 and miR-138-5p in regulating OSCC tumorigenesis and metastasis.

Article Snippet: Human OSCC cell lines SCC9, SCC15, and CAL27 were obtained from ATCC (Rockville, MD, USA).

Techniques: Expressing

Journal: BioMed Research International

Article Title: Aberrant Expression and Subcellular Localization of PER2 Promote the Progression of Oral Squamous Cell Carcinoma

doi: 10.1155/2020/8587458

Figure Lengend Snippet: PER2 expression and nuclear translocation are disturbed in OSCC tissues and adjacent noncancerous tissues. (a) Detection of PER2 expression levels on a tissue microarray. (b) Immunofluorescence staining for PER2 (green) in the 36 paired collected tissue samples and the 36 normal samples. Nuclei were stained by DAPI (blue). (c) The relative mRNA levels of PER2 in the 36 paired collected tissue samples and the 36 normal samples. (d and e) Representative western blotting analysis of PER2 in the 36 paired collected tissue samples and the 36 normal samples. GAPDH was used as the loading control (OSCC, oral squamous cell carcinoma tissues; ANT, adjacent noncancerous tissues; NT, normal oral tissues). ∗∗∗ P < 0.001 (compared with NT), # P < 0.05, and ### P < 0.001 (compared with ANT).

Article Snippet: Three human OSCC cell lines (SCC15, SCC25, and CAL27) obtained from ATCC were used for experiments.

Techniques: Expressing, Translocation Assay, Microarray, Immunofluorescence, Staining, Western Blot, Control

Journal: BioMed Research International

Article Title: Aberrant Expression and Subcellular Localization of PER2 Promote the Progression of Oral Squamous Cell Carcinoma

doi: 10.1155/2020/8587458

Figure Lengend Snippet: PER2 expression is remarkably downregulated and its nuclear translocation is blocked in OSCC cells. (a) Immunofluorescence staining for PER2 (green) in three OSCC cell lines and OKC cells. Nuclei were stained by DAPI (blue). (b) The PER2 protein expression levels of three OSCC cell lines in both cytoplasm and nucleus at each time point.

Article Snippet: Three human OSCC cell lines (SCC15, SCC25, and CAL27) obtained from ATCC were used for experiments.

Techniques: Expressing, Translocation Assay, Immunofluorescence, Staining

Journal: BioMed Research International

Article Title: Aberrant Expression and Subcellular Localization of PER2 Promote the Progression of Oral Squamous Cell Carcinoma

doi: 10.1155/2020/8587458

Figure Lengend Snippet: PER2 inhibits OSCC cell proliferation, migration, and invasion in vitro . (a) Western blotting analysis of PER2 expression. (b) The monoclone of PER2-overexpressed CAL27 cells. (c) PER2 inhibited cell proliferation of CAL27 cells. (d and e) Growth of OSCC cells was detected using the clone formation assay. (f and g) Representative microphotographs of wound-healing assays. Black lines mark the migration front. (h and i) Representative microphotographs of invasion assays.

Article Snippet: Three human OSCC cell lines (SCC15, SCC25, and CAL27) obtained from ATCC were used for experiments.

Techniques: Migration, In Vitro, Western Blot, Expressing, Tube Formation Assay

Journal: BioMed Research International

Article Title: Aberrant Expression and Subcellular Localization of PER2 Promote the Progression of Oral Squamous Cell Carcinoma

doi: 10.1155/2020/8587458

Figure Lengend Snippet: PER2 controls the EMT process in OSCC. (a-c) Quantitative RT-PCR analysis of EMT-associated genes in SCC15 (a), SCC25 (b), and CAL27 (c). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Three human OSCC cell lines (SCC15, SCC25, and CAL27) obtained from ATCC were used for experiments.

Techniques: Quantitative RT-PCR

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Long non-coding RNA P4713 contributes to the malignant phenotypes of oral squamous cell carcinoma by activating the JAK/STAT3 pathway

doi:

Figure Lengend Snippet: Analysis of long non-coding RNAs (lncRNAs) in oral squamous cell carcinoma (OSCC). A: Heat map, volcano plot, and log-log scatter plot showing differentially expressed lncRNAs between four OSCC samples and paired adjacent normal tissues (fold change > 2.0, P < 0.05). B: The expression of lncRNA P4713 was detected by quantitative real-time PCR (qRT-PCR) in 22 OSCC tissues and adjacent non-cancerous tissues. The results are expressed as log10 (2-ΔΔCt). A log2 fold change ≥ +2 or ≤ -2 was considered significant upregulation or downregulation (dotted lines). C: Relative P4713 expression in OSCC cell lines was measured by qRT-PCR. Columns represent the mean of three independent experiments; bars, the s.d; *P < 0.05; **P < 0.01. D: Confocal microscopic fluorescent in situ hybridization images and qRT-PCR results. Scale bar = 10 µm. E: Genome location analysis of human P4713 by the UCSC Genome Browser. F: Representative fluorescent images of at least three independent experiments. Scale bar = 10 µm. G: Relative levels of GFP expression by Western blot.

Article Snippet: Microarray analysis The OSCC lncRNA/mRNA microarray analysis was performed by CapitalBio Corporation (Beijing, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, In Situ Hybridization, Western Blot

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Long non-coding RNA P4713 contributes to the malignant phenotypes of oral squamous cell carcinoma by activating the JAK/STAT3 pathway

doi:

Figure Lengend Snippet: The effects of P4713 on oral squamous cell carcinoma cell proliferation in vitro. A: The relative expression of P4713 was examined by qRT-PCR in HSC-3 and UM1 cells. B: Cell proliferation was measured by CCK-8 assay. C: Detection for colony-formation assays after knockdown of P4713. D: Cell cycle analysis using propidium iodide staining. E: Western blot analysis of cyclin D1, CDK4, and CDK6 after P4713 knockdown.

Article Snippet: Microarray analysis The OSCC lncRNA/mRNA microarray analysis was performed by CapitalBio Corporation (Beijing, China).

Techniques: In Vitro, Expressing, Quantitative RT-PCR, CCK-8 Assay, Knockdown, Cell Cycle Assay, Staining, Western Blot

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Long non-coding RNA P4713 contributes to the malignant phenotypes of oral squamous cell carcinoma by activating the JAK/STAT3 pathway

doi:

Figure Lengend Snippet: Silencing of P4713 suppressed the migration and invasion of oral squamous cell carcinoma (OSCC) cells. A: Inhibition of migration in HSC-3 and UM1 cells after P4713 knockdown. B: A Matrigel invasion assay was performed using an invasion chamber after treatment with si-P4713. C: In vitro migration was assessed by wound healing experiments. D: E-cadherin, N-cadherin, and vimentin were analyzed by western blotting.

Article Snippet: Microarray analysis The OSCC lncRNA/mRNA microarray analysis was performed by CapitalBio Corporation (Beijing, China).

Techniques: Migration, Inhibition, Knockdown, Invasion Assay, In Vitro, Western Blot

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Long non-coding RNA P4713 contributes to the malignant phenotypes of oral squamous cell carcinoma by activating the JAK/STAT3 pathway

doi:

Figure Lengend Snippet: The relationship between P4713 and the JAK/STAT3 pathway. (A) Hierarchically clustered heatmaps of mRNAs altered in oral squamous cell carcinoma (OSCC; fold change > 2, P < 0.05). (B) The lncRNA-P4713 subnetwork in the OSCC co-expression network. (C) The top twenty GO terms of upregulated and downregulated mRNAs in OSCC cases (P < 0.05). (D) Significantly enriched pathways of the indicated gene sets. (E) qRT-PCR and (F) western blotting were used to measure the JAK/STAT3 pathway affected by P4713. (G) Representative fluorescent images of the location of STAT3. Scale bar = 10 µm.

Article Snippet: Microarray analysis The OSCC lncRNA/mRNA microarray analysis was performed by CapitalBio Corporation (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: Mitochondrial proteins upregulated in STAT3-low versus STAT3-high samples in proteomic shotgun analysis ( n = 4). The y-axis shows the p value as −log10 and the x-axis indicates the difference in fold change (FC) between low vs. high STAT3 samples. Highlighted are NDUFS1 from complex I (orange) and ATP5O, ATP5A1 and ATP5C1 from complex V (blue). The dotted line represents level of 0.01 significance.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques:

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: Clinical characteristics of the PCa patient cohort stained for NDUFS1 and ATPO.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Staining

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) NDUFS1 immunohistochemistry staining of PCa tissue microarray (TMA) samples. (0) staining intensity of 0 to 0.5, (1) staining intensity of 1, (2) staining intensity of 2, (3) staining intensity of 3, (control) technical negative control staining (left). Scale bar = 100 µm. ( B ) ATP5O immunohistochemistry staining of PCa TMA samples. (0) staining intensity of 0 to 0.5, (1) staining intensity of 1, (2) staining intensity of 2, (3) staining intensity of 3, (control) technical negative control staining (right). Scale bar = 100 µm.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Immunohistochemistry, Staining, Microarray, Control, Negative Control

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) IHC staining intensities for NDUFS1 and ATP5O ranging from 0 to 3 in PCa samples and adjacent benign tissue. Boxplots show the interquartile range with horizontal line as median, whiskers as upper and lower limits. ( B ) Histogram of staining intensities for NDUFS1 and ATP5O ranging from 0 to 3 in PCa samples. Value 0 includes staining of 0 and 0.5, value 1 includes staining of 1 and 1.5, and value 2 includes staining of 2 and 2.5.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Immunohistochemistry, Staining

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) Kaplan-Meier plot showing time to biochemical recurrence in months for NDUFS1 in the IHC cohort ( n = 60). p -values were estimated by log-rank test. Hazard ratio of 2.188 with a confidence interval of 1.056 to 4.530. Blue = low expression, orange = high expression. ( B ) Kaplan-Meier plot showing time to biochemical recurrence in months for ATP5O in the IHC cohort ( n = 61). p -values were estimated by log-rank test. Hazard ratio of 2.953 with a confidence interval of 1.364 to 6.391. Blue = low expression, orange = high expression.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Expressing

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) NDUFS1 mRNA expression levels analyzed from prostate carcinoma epithelia ( n = 30; 1.74-fold increase), prostatic intraepithelial neoplasia epithelia ( n = 13; 1.69-fold increase), or normal prostate gland samples ( n = 21). ( B ) ATP5O mRNA expression levels analyzed from prostate carcinoma epithelia ( n = 30; 1.83-fold increase), prostatic intraepithelial neoplasia epithelia ( n = 13; 3.01-fold increase), or normal prostate gland samples ( n = 23). ( C ) NDUFS1 mRNA expression levels (TPM) analyzed from TCGA-PRAD primary tumor tissue ( n = 301) and true normal tissue samples (cancer-free patients) from GTEx ( n = 100). ( D ) ATP5O mRNA expression levels (TPM) analyzed from TCGA-PRAD primary tumor tissue ( n = 301) and true normal tissue samples (cancer-free patients) from GTEx ( n = 100). Data for ( A,B ) were extracted from the Oncomine™ Platform from the Tomlins Prostate dataset. Data for ( C – D ) were extracted from The Cancer Genome Atlas-Prostate Adenocarcinoma data collection and the Genotype-Tissue Expression project. Representation: boxes as interquartile range, horizontal line as the mean, whiskers as lower and upper limits.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Expressing

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) NDUFS1 mRNA expression levels in prostate cancer patients, analyzed from primary PCa ( n = 10) and metastasis ( n = 4; 3.00-fold increase). ( B ) ATP5O mRNA expression levels in prostate cancer patients, analyzed from primary PCa ( n = 8) and metastasis ( n = 3; 1.75-fold increase). Data were extracted from the Oncomine™ Platform from the ( A ) Ramaswamy Multi-cancer and ( B ) Magee Prostate datasets. Representation: boxes as interquartile range, horizontal line as the mean, whiskers as lower and upper limits.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Expressing

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) NDUFS1 mRNA expression levels in prostate cancer patients, analyzed from hormone naive ( n = 3) and hormone refractory ( n = 13; 2.64-fold increase) tumors. ( B ) ATP5O mRNA expression levels in prostate cancer patients, analyzed from hormone naive ( n = 3) and hormone refractory ( n = 16; 1.56-fold increase) tumors. Data for ( A , B ) were extracted from the Oncomine™ Platform from the Tomlins Prostate dataset. Representation: boxes as interquartile range, horizontal line as the mean, whiskers as lower and upper limits.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Expressing

Journal: Cancers

Article Title: Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer

doi: 10.3390/cancers13236036

Figure Lengend Snippet: ( A ) NDUFS1 mRNA expression levels (TPM) analyzed from TCGA-PRAD primary tumor tissue ( n = 301) and adjacent normal tissue ( n = 52). ( B ) ATP5O mRNA expression levels (TPM) analyzed from TCGA-PRAD primary tumor tissue ( n = 301) and adjacent normal tissue ( n = 52). Data for ( A , B ) were extracted from The Cancer Genome Atlas-Prostate Adenocarcinoma data (TCGA-PRAD) collection. Representation: boxes as interquartile range, horizontal line as the mean.

Article Snippet: The following antibodies were used: anti-NDUFS1 antibody (rabbit polyclonal, 1:100 dilution; abcam; ab169540) and anti-ATP5O antibody (rabbit polyclonal, 1:200 dilution; proteintech, 10994-1-AP).

Techniques: Expressing