Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Association between the clinicopathological variables and BTC expression in 38 OSCC patients.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing, Virus, Infection

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: DEGs in OSCC. (A,B) Volcano plots were constructed using FC values and FDRs. The red points in the plot represent the overexpressed mRNAs, and the blue points indicate the downregulated mRNAs with statistical significance. (A) DEGs between normal and tumor tissues. (B) DEGs between metastasis-positive and metastasis-negative tumor tissues. (C) Hierarchical clustering analysis of mRNAs that were differentially expressed between metastasis-negative and metastasis-positive tissues. The normalized expression levels in the heatmaps are colored from blue to red in ascending order. (D) Multivariate Cox regression analysis according to gene expression. (E) Kaplan–Meier survival curves were performed to show the prognosis of patients with high and low expression of BTC through the analysis of the mRNA expression profile data of 260 OSCC tumor samples from TCGA database.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Construct, Expressing, Gene Expression

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: BTC expression is correlated with clinicopathological parameters in OSCC patients. (A) Representative images of immunohistochemical staining and (B) immunoreactive score of BTC in human normal mucosa samples, OSCC tissue samples, and metastatic LN samples. The experiment was repeated three times independently. Results are shown as mean ± SD. T-test, n = 38. (C) mRNA expression of BTC in normal and tumor tissues was detected by RT-PCR. (D) Kaplan–Meier survival curves of 38 patients from our department. (E–L) Analysis of 330 OSCC samples from TCGA database and 32 pairs of OSCC samples selected from TCGA database showed the comparison of (E,F) BTC expressed in tumor tissues and normal tissues. Correlation analysis with BTC expression and (G) age, (H) histological grade, (I) sex, (J) T category, (K) tumor stage, and (L) LN metastasis status. * p < 0.05. ** p < 0.01. *** p < 0.001. **** p < 0.0001.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Reverse Transcription Polymerase Chain Reaction, Comparison

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Statistical analyses of clinicopathological features associated with survival in 38 OSCC patients with the multivariate Cox proportional hazards models.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Overexpression of BTC inhibits the proliferation, migration, and invasion of OSCC cell lines. (A) Cancer cell transfectants of the BTC-expressing vector and empty vector control were identified in SCC4 and CAL27 cells by Western blot. (B,C) Overexpression of BTC inhibited cell proliferation, as indicated by the CCK-8 assay, in CAL27 and SCC4 cells. (D) Wound healing assay showed that overexpression of BTC inhibited CAL27 and SCC4 cell migration. (E,F) Transwell assays showed that the migration and invasion abilities of CAL27 and SCC4 cells were impaired after overexpression of BTC.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Over Expression, Migration, Expressing, Plasmid Preparation, Control, Western Blot, CCK-8 Assay, Wound Healing Assay

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: (A) Comparison of EMT scores in normal and tumor (in BTC low- and high-expression groups) tissue. (B) TCGA and GSEA showed highly regulated genes in patients with high-BTC expression versus those with low-BTC expression. (C) Heatmap of EMT marker expression in the BTC high- and low-expression groups. (D–F) Relationship between BTC and EMT markers, such as E-cadherin, N-cadherin, and vimentin. (G) PPI network analysis and Western blot analysis. The PPI network of the DEGs was constructed using STRING. The network nodes represent different proteins. The edges represent protein–protein associations, and the line thickness indicates the strength of the supporting data. (H) Protein expression level of EMT-related markers and the PI3K-AKT signaling pathway after overexpression of BTC in SCC4 and Cal27 cells.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Comparison, Expressing, Marker, Western Blot, Construct, Over Expression

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: The expression of circ_0000140 in OSCC tissues and cells. a The expression of circ_0000140 in OSCC tissues (OSCC) and adjacent normal mucosal tissues (Normal) was detected by qRT-PCR. b The circ_0000140 expression in OSCC tissues with or without lymph node metastasis (Yes or No) was measured by qRT-PCR. c qRT-PCR was used to test the circ_0000140 expression in OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) and HOK cells. d , e The relative expression levels of circ_0000140 and KIAA0907 in CAL-27 and SCC-4 cells were assessed by qRT-PCR after treatment with RNase R. f , g The relative expression levels of circ_0000140, U6 and 18 s rRNA in the nuclear and cytoplasmic of CAL-27 and SCC-4 cells were detected by qRT-PCR. * P < 0.05, ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: Effects of circ_0000140 overexpression on the proliferation, migration and invasion of OSCC cells. CAL-27 and SCC-4 cells were transfected with circ_0000140 overexpression vector or vector. a The expression of circ_0000140 in CAL-27 and SCC-4 cells was detected by qRT-PCR to evaluate transfection efficiency. b Colony formation assay was performed to measure the number of colonies in CAL-27 and SCC-4 cells. c , d The number of migrated and invaded CAL-27 and SCC-4 cells was determined by transwell assay. e , f WB analysis was performed to detect the protein levels of ki67, MMP-2 and MMP-9 in CAL-27 and SCC-4 cells. ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Over Expression, Migration, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Colony Assay, Transwell Assay

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: Effects of circ_0000140 overexpression on the glycolysis metabolism of OSCC cells. CAL-27 and SCC-4 cells were transfected with circ_0000140 overexpression vector or vector. a , b The ECAR of CAL-27 and SCC-4 cells was measured by Seahorse XF Extracellular Flux Analyzer. c The lactate acid level of CAL-27 and SCC-4 cells was tested by Lactate Assay Kit. d , e The protein levels of GLUT1 and LDHA in CAL-27 and SCC-4 cells were detected by WB analysis. ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Over Expression, Transfection, Plasmid Preparation, Lactate Assay

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: Circ_0000140 could absorb miR-182-5p. a The sequences of circ_0000140 containing the miR-182-5p binding sites or mutant binding sites were presented. b , c Dual-luciferase reporter assay was used to detect the interaction between circ_0000140 and miR-182-5p in CAL-27 and SCC-4 cell. d , e The enrichment of circ_0000140 and miR-182-5p in anti-Ago2 or anti-IgG was measured by the RIP assay. f qRT-PCR was performed to measure the expression of miR-182-5p in OSCC cells (CAL-27 and SCC-4) and NOK cells. g The expression of miR-182-5p in CAL-27 and SCC-43 cells was assessed by qRT-PCR to assess the effect of circ_0000140 overexpression on miR-182-5p expression. * P < 0.05, ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Quantitative RT-PCR, Expressing, Over Expression

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: Effects of miR-182-5p mimic on the progression of OSCC cells. a The expression of miR-182-5p in CAL-27 and SCC-4 cells was measured by qRT-PCR to evaluate the transfection efficiency of miR-182-5p mimic. CAL-27 and SCC-4 cells were co-transfected with circ_0000140 overexpression vector and miR-182-5p mimic. b The number of colonies in CAL-27 and SCC-4 cells was detected by colony formation assay. c , d The number of migrated and invaded CAL-27 and SCC-4 cells was tested by transwell assay. e , f The protein levels of ki67, MMP-2 and MMP-9 in CAL-27 and SCC-4 cells were assessed by WB analysis. g Seahorse XF Extracellular Flux Analyzer was used to evaluate the ECAR of CAL-27 and SCC-4 cells. h The lactate acid level of CAL-27 and SCC-4 cells was determined by Lactate Assay Kit. i , j WB analysis was performed to measure the protein levels of GLUT1 and LDHA in CAL-27 and SCC-4 cells. ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Colony Assay, Transwell Assay, Lactate Assay

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: MiR-182-5p could target CDC73. a The sequences of CDC73 3′UTR containing the miR-182-5p binding sites or mutant binding sites were shown. b , c The interaction between miR-182-5p and CDC73 in CAL-27 and SCC-4 cells was assessed by dual-luciferase reporter assay. d The protein level of CDC73 in OSCC cells (CAL-27 and SCC-4) and HOK cells was determined by WB analysis. e WB analysis was used to measure the CDC73 protein level in CAL-27 and SCC-4 cells to evaluate the miR-182-5p expression on CRC73 expression. f , g The CDC73 protein level in CAL-27 and SCC-4 cells was detected by WB analysis to evaluate the circ_0000140 and miR-182-5p expression on CDC73 expression. ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Expressing

Journal: Cancer Cell International

Article Title: Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p

doi: 10.1186/s12935-020-01501-7

Figure Lengend Snippet: Effects of CDC73 silencing on the progression of OSCC cells. a The protein level of CDC73 in CAL-27 and SCC-4 cells was measured by WB analysis to evaluate the transfection efficiency of si-CDC73. CAL-27 and SCC-4 cells were co-transfected with anti-miR-182-5p and si-CDC73. b Colony formation assay was performed to assess the number of colonies in CAL-27 and SCC-4 cells. c The number of migrated and invaded CAL-27 and SCC-4 cells was detected by transwell assay. d WB analysis was used to measure the protein levels of ki67, MMP-2 and MMP-9 in CAL-27 and SCC-4 cells. e , f Seahorse XF Extracellular Flux Analyzer was employed to assess the ECAR of CAL-27 and SCC-4 cells. g The lactate acid level of CAL-27 and SCC-4 cells was detected by Lactate Assay Kit. h The protein levels of GLUT1 and LDHA in CAL-27 and SCC-4 cells were tested by WB analysis. ** P < 0.01

Article Snippet: OSCC cell lines (CAL-27, SCC-4, SCC-9 and SCC-25) were bought from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Transfection, Colony Assay, Transwell Assay, Lactate Assay

Journal: OncoTargets and therapy

Article Title: Androgen receptor promotes oral squamous cell carcinoma cell migration by increasing EGFR phosphorylation

doi: 10.2147/OTT.S200718

Figure Lengend Snippet: Expression of AR, EGFR, and pEGFR in OSCC tumors. ( A ) AR-positive staining in OSCC tumor (red arrow: cancer cell). ( B ) AR-positive staining in OSCC tumor. ( C ) AR staining in prostate cancer. ( D and E ) tEGFR staining in AR-positive OSCC tumor and AR-negative OSCC tumor. ( F ) No difference in tEGFR expression between AR-positive tumors and AR-positive tumors (* p >0.05). ( G and H ) pEGFR staining in AR-positive OSCC tumor and AR-negative OSCC tumor. ( I ) AR-positive tumors express significantly higher pEGFR ( # p <0.01). Scale bar in A – E, G , and H , 50 µm. Abbreviations: AR, androgen receptor; OSCC, oral squamous cell carcinoma; EGFR, epidermal growth factor receptor; pEGFR, phosphorylated EGFR; tEGFR, total EGFR.

Article Snippet: As shown in , four OSCC cell lines (SAS, SCC4, SCC9, and OECM-1) expressed different levels of AR mRNA, which were lower than that in LNCaP cells.

Techniques: Expressing, Staining

Journal: OncoTargets and therapy

Article Title: Androgen receptor promotes oral squamous cell carcinoma cell migration by increasing EGFR phosphorylation

doi: 10.2147/OTT.S200718

Figure Lengend Snippet: Expression of AR in different OSCC cell lines and LNCaP cells. ( A ) AR mRNA in LNCaP was used as a positive control, AR mRNA was detected in four OSCC cell lines, not SCC25. ( B ) Different levels of AR proteins expressed in these cell lines. Abbreviations: AR, androgen receptor; OSCC, oral squamous cell carcinoma.

Article Snippet: As shown in , four OSCC cell lines (SAS, SCC4, SCC9, and OECM-1) expressed different levels of AR mRNA, which were lower than that in LNCaP cells.

Techniques: Expressing, Positive Control

Journal: Oncotarget

Article Title: WISP-1, a novel angiogenic regulator of the CCN family, promotes oral squamous cell carcinoma angiogenesis through VEGF-A expression

doi:

Figure Lengend Snippet: (A) SCC4 cells stably expressing shRNA constructs or control shRNA were seeded as monolayers and counted daily. Cells (10 3 ) were plated in 6 well plates and grown for 2 days. Cells were trypsinized, and cell numbers was counted. (B–C) WISP-1 and VEGF-A mRNA and protein expression in SCC4 cells stably expressed a control shRNA or a WISP-1 shRNA was examined by western blot, qPCR, and ELISA. (D–E) EPCs were incubated with CM collected from control-shRNA and WISP-1-shRNA transfected SCC4 cells for 24 h and cell migration or tube formation were examined. (F) PBS, VEGF-A, control shRNA/SCC4 CM, and WISP-1 shRNA/SCC4 CM mixed in Matrigel were placed on chick chorioallantoic membranes. CAMs in each group were photographed on developmental day 12. (G) Mice were subcutaneously injected with Matrigel mixed with PBS, control shRNA/SCC4 CM or WISP-1 shRNA/SCC4 CM for seven days. Plugs excised from the mice were photographed and stained with CD31. (H) Control shRNA and WISP-1 shRNA SCC4 cells were mixed with Matrigel and injected into the flank of the mice for 28 days. Tumor growth was monitored using the IVIS Imaging System. Tumor growth was quantified by fluorescent imaging from week 0–6. (I) Tumors were paraffin embedded, and sections were immunostained using the WISP-1, VEGF-A, and CD31 antibodies. (E = epithelial, T = tumor, S = stroma). (J) Diagrammatic model for the role of WISP-1 in OSCC. (1) WISP-1 induces VEGF-A expression and secretion in OSCC cells through the integrin αvβ3/FAK/c-Src pathway, which transactivates the EGFR/ERK/HIF1-α signal pathway. (2) The WISP-1-induced secretion of VEGF-A subsequently recruiting EPCs to OSCC tumor microenvironment and promoting neoangiogenesis. Data represent the mean ± SEM * P < 0.05 compared to control shRNA/SCC4.

Article Snippet: Moreover, our result indicated that OSCC cell lines (SCC4 and SAS) and OSCC tumor specimens showed highly expression of VEGF-A protein compared with normal specimens ( ).

Techniques: Stable Transfection, Expressing, shRNA, Construct, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Transfection, Migration, Injection, Staining, Imaging