Journal: Molecular Therapy. Nucleic Acids

Article Title: m6A demethylase FTO suppresses pancreatic cancer tumorigenesis by demethylating PJA2 and inhibiting Wnt signaling

doi: 10.1016/j.omtn.2021.06.005

Figure Lengend Snippet: Relative N6-methyladenine (m6A) RNA modification in total RNA is upregulated in pancreatic cancer (PC) (A and B) m6A content in total RNA in 16 pairs of pancreatic adenocarcinoma (PAAD) and corresponding para-tumor tissues. Except for patients 3 and 8, the relative m6A RNA modification of PAAD tissues was significantly higher than that of the corresponding para-tumor tissues. (C) Relative m6A RNA modification was higher in PC cell lines. Human PC cell lines (SW1990, AsPC-1, PANC-1, BxPC-3, and Capan-2) and a human immortalized pancreatic duct epithelial cell line (HPDE6-C7) were subjected to RNA m6A detection. Tumor, PAAD tissue. (B) ∗p < 0.05 by paired Student’s t test; (C) ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by unpaired Student’s t test. Data are represented as mean ± standard error.

Article Snippet: Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCC CRL-1682), PANC-1 (ATCC CRL-1469), BxPC-3 (ATCC CRL-1687), and Capan-2 (ATCC HTB-80) were purchased from ATCC (Manassas, VA, USA).

Techniques: RNA modification

Journal: Molecular Therapy. Nucleic Acids

Article Title: m6A demethylase FTO suppresses pancreatic cancer tumorigenesis by demethylating PJA2 and inhibiting Wnt signaling

doi: 10.1016/j.omtn.2021.06.005

Figure Lengend Snippet: FTO acts as m6A demethylase in pancreatic cell lines (A and B) Quantitative real-time PCR and WB assays showed reduced FTO levels in five PC cell lines. The mRNA and protein levels of FTO were (C and E) significantly reduced in SW1990 cells after small interfering (si)RNAs (siFTO-1, siFTO-2, and siFTO-3) transfection, whereas were (D and F) significantly increased in BxPC-3 cells after transfection with wild-type FTO (FTO-WT) and mutant FTO (FTO-mut) overexpressing plasmids. (G) FTO knockdown significantly upregulated the m6A level in total RNA of SW1990 cells. (H) FTO-WT overexpression significantly decreased the m6A level in total RNA of BXPC-3 cells. No significant changes were observed in the m6A levels upon FTO-mut overexpression. GAPDH was used as internal control in the quantitative real-time PCR and WB assays. NC, negative control. (A–H) ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, no statistical significance by unpaired Student’s t test.

Article Snippet: Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCC CRL-1682), PANC-1 (ATCC CRL-1469), BxPC-3 (ATCC CRL-1687), and Capan-2 (ATCC HTB-80) were purchased from ATCC (Manassas, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Transfection, Mutagenesis, Knockdown, Over Expression, Control, Negative Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: m6A demethylase FTO suppresses pancreatic cancer tumorigenesis by demethylating PJA2 and inhibiting Wnt signaling

doi: 10.1016/j.omtn.2021.06.005

Figure Lengend Snippet: FTO enhanced PJA2 stability in an m6A-YTHDF2-dependent manner in PC cells (A and B) Venn diagrams illustrated overlap in differentially bioinformatics analyses of the top 20 genes significantly positively or negatively related to FTO in PC. (C–F) Quantitative real-time PCR was performed to detect the mRNA expression of eight candidate genes after knockdown and overexpression of FTO in BxPC-3 and SW1990 cells. Only the PJA2 expression pattern, by both quantitative real-time PCR and WB, was consistent with that of FTO. (G) cBioPortal analysis displayed the co-expression relationship between FTO and PJA2 in PAAD tissues. Spearman coefficient: 0.74 (p = 2.10e−32); Pearson coefficient: 0.75 (p = 7.98e−34). (H and I) The relative PJA2 m6A level was detected in BxPC-3 and SW1990 cells after knockdown and overexpression of FTO, respectively. (J–M) Quantitative real-time PCR and WB were performed to detect the mRNA and protein expression of PJA2 after YTHDF1 and YTHDF2 knockdown by siRNA in BxPC-3 and SW1990 cells. (N and O) PJA2 stability was detected after knocking down YTHDF2, FTO by siRNA, and overexpression of FTO-WT and FTO-mut in BxPC-3 and SW1990 cells. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

Article Snippet: Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCC CRL-1682), PANC-1 (ATCC CRL-1469), BxPC-3 (ATCC CRL-1687), and Capan-2 (ATCC HTB-80) were purchased from ATCC (Manassas, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Knockdown, Over Expression

Journal: Frontiers in Oncology

Article Title: The Role of Long Noncoding RNA AL161431.1 in the Development and Progression of Pancreatic Cancer

doi: 10.3389/fonc.2021.666313

Figure Lengend Snippet: Knockdown of lncRNA AL161431.1 inhibited the growth, invasion and migration and promoted apoptosis in pancreatic adenocarcinoma cells. (A) Relative AL161431.1 expression in PANC-1, SW1990, and BxPC-3 cells analyzed by qRT-PCR; (B) Relative AL161431.1 expression in SW1990 cells after siRNA transfection analyzed by qRT-PCR; (C) CCK-8 assay showing fold change at OD450, representing growth curve of SW1990 and BxPC-3 cells at 24, 48, and 72 hours after siRNA1 transfection; (D) Wound healing assay in naïve, scramble siRNA, or siRNA1 transfected SW1990 or BxPC-3 cells at 0, 24, and 48 hours post scratching (100x); (E) Transwell assay in scrambled control siRNA or siRNA1 transfected SW-1990 or BxPC-3 cells (100x); (F) Cell death (Q2+Q3) in scrambled control siRNA or siRNA1 transfected SW1990 and BxPC-3 cells analyzed by flow cytometry; (G) Cell cycle analysis of SW1990 and BxPC-3 cells after transfection of scrambled control siRNA or siRNA1 by flow cytometry. NC: scrambled control siRNA or shRNA. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A specific SW1990 cell line (SW1990-LNC-KD) with stable knockdown of lncRNA AL161431.1 or control SW1990 cell line was constructed using a lnc-shRNA sequence targeting 5’- GCAGTATTCCTGCACTTCT -3’ or scramble control sequence 5’- TTCTCCGAACGTGTCACGT -3’ cloned into the pLV3(H1/GFP&Puro) vector , and transfected into 293T cells (Shanghai GenePharma China).

Techniques: Migration, Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Wound Healing Assay, Transwell Assay, Flow Cytometry, Cell Cycle Assay, shRNA

Journal: Frontiers in Oncology

Article Title: The Role of Long Noncoding RNA AL161431.1 in the Development and Progression of Pancreatic Cancer

doi: 10.3389/fonc.2021.666313

Figure Lengend Snippet: Knockdown of lncRNA AL161431.1 inhibited the growth of tumor xenografts. (A) Size of SW1990 tumor xenografts with SW1990-LNC-KD or SW1990-LNC-NC cells at 14 days after injection; (B) Time course of in vivo xenografts growth measured in mm 3 ; (C) Relative AL161431.1 expression in engrafted tumors with SW1990-LNC-NC cell or SW1990-LNC-KD cell; (D) HE staining of tumor xenografts; (E) PCNA immunohistochemistry in tumor xenografts; (F) Relative ratio of PCNA positive cells in the engrafted tumors with SW1990-LNC-NC cell or SW1990-LNC-KD cell. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A specific SW1990 cell line (SW1990-LNC-KD) with stable knockdown of lncRNA AL161431.1 or control SW1990 cell line was constructed using a lnc-shRNA sequence targeting 5’- GCAGTATTCCTGCACTTCT -3’ or scramble control sequence 5’- TTCTCCGAACGTGTCACGT -3’ cloned into the pLV3(H1/GFP&Puro) vector , and transfected into 293T cells (Shanghai GenePharma China).

Techniques: Injection, In Vivo, Expressing, Staining, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: The Role of Long Noncoding RNA AL161431.1 in the Development and Progression of Pancreatic Cancer

doi: 10.3389/fonc.2021.666313

Figure Lengend Snippet: Knockdown of lncRNA AL161431.1 inhibited EMT. (A) qRT-PCR showing the knock down of lncRNA AL161431.1, and the corresponding changes in E-cadherin, N-cadherin, and vimentin in SW1990 and BxPC-3 cells; (B) Western blots showing the changes in protein level in E-cadherin, N-cadherin, and vimentin in SW1990 and BxPC-3 cells after transfection of scrambled siRNA or siRNA1; (C) Immunofluorescence analysis showing the change of expression of E-cadherin, N-cadherin, and vimentin in SW1990 and BxPC-3 cells after transfection of scrambled siRNA or siRNA1 (200x). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A specific SW1990 cell line (SW1990-LNC-KD) with stable knockdown of lncRNA AL161431.1 or control SW1990 cell line was constructed using a lnc-shRNA sequence targeting 5’- GCAGTATTCCTGCACTTCT -3’ or scramble control sequence 5’- TTCTCCGAACGTGTCACGT -3’ cloned into the pLV3(H1/GFP&Puro) vector , and transfected into 293T cells (Shanghai GenePharma China).

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Immunofluorescence, Expressing

Journal: Frontiers in Pharmacology

Article Title: Phytopharmacology and Clinical Updates of Berberis Species Against Diabetes and Other Metabolic Diseases

doi: 10.3389/fphar.2020.00041

Figure Lengend Snippet: In vitro activity of extracts and/or isolated compounds from Berberis species against diabetes and metabolic diseases.

Article Snippet: Berberine (BBR) , CEM, HCT-116, HepG2.2.15, SW1990, HT1080 and 293T cell lines , ↑gene expression of the insulin receptor , ( ) .

Techniques: In Vitro, Activity Assay, Isolation, Binding Assay, Activation Assay, Cell Culture, Expressing, Small Interfering RNA, Blocking Assay, Gene Expression, Inhibition, Translocation Assay, Phospho-proteomics, Modification

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: miR-142-3p attenuates the sensitivity to irradiation in CRC cells. ( A ) The TCGA dataset showed the expression of miR-142-3p in COAD (colorectal adenocarcinoma). ( B ) Left: Representative images of colony formation in SW480 and SW620 cells irradiated at doses of 0, 3, 6, and 9 Gy. Right: Surviving fraction of CRC cells. ( C ) qPCR analysis confirmed the stable overexpression of miR-142-3p in SW480 and SW620 cells. ( D ) Upper panel: Representative images of colony formation in the cancer cell lines after 6 Gy irradiation (NC, LV, miR-142-3p). Lower panel: Quantification of colony formation. ( E ) The CCK-8 assay revealed proliferation curves of SW480 and SW620 cells (NC, LV, miR-142-3p) after 6 Gy. ( F ) γH2AX was detected in NC, LV, and miR-142-3p cells after irradiation treatment for 72 h using immunofluorescence analysis. Scale bar = 20 μm. The bar graph showed the average number of γH2AX positive cells per field. ( G ) Western blot showed γH2AX expression under 0–6 Gy of irradiation in SW480 and SW620 cells (NC, LV, miR-142-3p). β-actin was used as the internal control. All data were represented as mean ± SD. ns = not significant; **** P < 0.0001. ( n = 3 per group)

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: Irradiation, Expressing, Over Expression, CCK-8 Assay, Immunofluorescence, Western Blot, Control

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: FOXO4 is a direct target of miR-142-3p. ( A ) TargetScan predicted a binding site between miR-142-3p and the 3’ UTR of FOXO4. The schematic diagram showed the complementary base pairing between miR-142-3p and the 3’ UTR of FOXO4 (positions 685-692 of the FOXO4 3’ UTR). ( B ) The expression of FOXO4 in the COAD dataset (TCGA). ( C ) A schematic illustration of a potential miR-142-3p binding site in the FOXO4 3’ UTR. The MT sequence of the FOXO4 3’ UTR was presented alongside the WT sequence. ( D ) A dual-luciferase assay was performed after transfecting the WT or MT construct into SW480 cells. After incubation for 48 h, relative luciferase activity was measured using renilla luciferase as a normalization control. ( E ) FOXO4 expression levels in SW480 and SW620 cells (NC, LV, miR-142-3p) were measured using qPCR. ( F ) FOXO4 expression levels in SW480 and SW620 cells (NC, LV, miR-142-3p) were measured using Western blot. β-actin was used as the loading control. All data were represented as mean ± SD. ns = not significant; **** P <0.0001. ( n = 3 per group)

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: Binding Assay, Expressing, Sequencing, Luciferase, Construct, Incubation, Activity Assay, Control, Western Blot

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: Inhibiting FOXO4 reduces radiosensitivity in CRC. ( A ) FOXO4 mRNA expression levels in SW480 and SW620 cells (NC, si-NC, si-FOXO4) were measured by qPCR. ( B ) FOXO4 and γH2AX protein expression levels in SW480 and SW620 cells (NC, si-NC, si-FOXO4) were measured by Western blot. β-actin was used as the loading control. ( C ) Upper panel: Representative images of colony formation in the cancer cell lines after 6 Gy irradiation (NC, si-NC, si-FOXO4). Lower panel: Quantification of colony formation. ( D ) The CCK-8 assay showed the proliferation curves of SW480 and SW620 cells (NC, si-NC, si-FOXO4) after 6 Gy. All data were represented as mean ± SD. ns = not significant; **** P < 0.0001. ( n = 3 per group)

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: Expressing, Western Blot, Control, Irradiation, CCK-8 Assay

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: Overexpression of FOXO4 can reverse the effect of miR-142-3p in promoting radioresistance in CRC. ( A ) FOXO4 mRNA expression levels in SW480 and SW620 cells (miR-142-3p, miR-142-3p + NC, miR-142-3p+FOXO4) were measured by qPCR. ( B ) FOXO4 and γH2AX protein expression levels in SW480 and SW620 cells (miR-142-3p, miR-142-3p + NC, miR-142-3p+FOXO4) were measured by Western blot. β-actin was used as the loading control. ( C ) Upper panel: Representative images of colony formation in the cancer cell lines after 6 Gy irradiation (miR-142-3p, miR-142-3p + NC, miR-142-3p+FOXO4). Lower panel: Quantification of colony formation. ( D ) The CCK-8 assay showed the proliferation curves of SW480 and SW620 cells (miR-142-3p, miR-142-3p + NC, miR-142-3p+FOXO4) after 6 Gy. All data were represented as mean ± SD. ns = not significant; **** P < 0.0001. ( n = 3 per group)

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: Over Expression, Expressing, Western Blot, Control, Irradiation, CCK-8 Assay

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: miR-142-3p activates NHEJ by targeting FOXO4, thereby promoting DNA damage repair. ( A ) Advanced volcano plot of RNA-seq comparing miR-142-3p over-expression cells with LV control cells. ( B , C ) KEGG Enrichment Bubble Plot and KEGG Enrichment Scatter Plot of RNA-seq comparing miR-142-3p over-expression cells with LV control cells. ( D ) GO Enrichment Scatter Plot of RNA-seq comparing miR-142-3p overexpression cells with LV control cells. ( E ) Western blot analysis of DNA damage response proteins (DNA-PKcs, Ku80, RAD50, MRE11, NBS1, XRCC1, LIG3) in SW480 and SW620 cells under different treatments (NC, LV, si-FOXO4, miR-142-3p, miR-142-3p+FOXO4). β-actin was used as the loading control. ( F ) Western blot analysis of γH2AX in SW480 and SW620 cells treated with 6 Gy of radiation. β-actin was used as the loading control

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: RNA Sequencing, Over Expression, Control, Western Blot

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: miR-142-3p confers resistance to radiation in a CRC xenograft model. ( A ) Schematic diagram of radiation exposure in nude mice with transplanted tumors. ( B ) Changes in mouse body weight: Body weights of mice bearing SW480 and SW620 tumors were measured every 3 days after tumor implantation. Data were presented as mean ± SD ( n = 5 per group). ( C , D ) Tumor volume growth curves: Tumor volumes of SW480 and SW620 xenografts in mice treated with miR-142-3p, LV, miR-142-3p + IR, and LV + IR were shown. ( E , F ) Representative images of tumors: SW480 and SW620 tumors harvested at the endpoint. ( G , H ) Final tumor weights: Quantitative analysis of tumor weights from SW480 and SW620 xenografts at the endpoint. ( I ) FOXO4 IHC staining and quantification: Representative IHC images (scale bar = 100 μm) of FOXO4 in SW480/SW620 xenograft tumors under different treatments (miR-142-3p, LV). Statistical significance; ** P < 0.01, *** P < 0.001. ( n = 3 per group)

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: Tumor Implantation, Immunohistochemistry

Journal: Biology Direct

Article Title: MiR-142-3p promotes radioresistance by targeting FOXO4 in colorectal cancer

doi: 10.1186/s13062-026-00772-w

Figure Lengend Snippet: miR-142-3p promotes DNA repair in CRC xenograft models, thereby reducing the efficacy of radiotherapy. ( A ) Ki67 IHC staining and quantification: Representative IHC images (scale bar = 100 μm) of Ki67 in SW480 and SW620 xenograft tumors under different treatments (miR-142-3p, miR-142-3p + IR, LV, LV + IR). Right panel: Quantification of Ki67-positive cells (%). All data were represented as mean ± SD. ns = not significant; **** P < 0.0001. ( n = 3 per group) ( B ) Hematoxylin and eosin (HE) staining: Representative HE images (scale bar = 100 μm) of SW480 and SW620 xenograft tumors under different treatments (miR-142-3p, miR-142-3p + IR, LV, LV + IR), showing histological changes (cell density, morphology). ( C ) IHC staining of γH2AX: Representative IHC images (scale bar = 100 μm) of γH2AX in SW480 and SW620 xenograft tumors under various treatments

Article Snippet: SW480 and SW620, human CRC cell lines, were purchased from Genechem Company (Shanghai, PR China) and BeNa Culture Collection (Beijing City, PR China).

Techniques: Immunohistochemistry, Staining

Journal: Discover Oncology

Article Title: Identification of the disulfidptosis-related gene G6PD as a potential biomarker and therapeutic target in pancreatic ductal adenocarcinoma by integrative bioinformatics analysis and experimental validation

doi: 10.1007/s12672-026-04753-3

Figure Lengend Snippet: G6PD Inhibitor RRx-001 Activates Disulfide-Mediated Cell Death in SW1990 Cells. A Molecular docking and optimization of RRx-001 with G6PD. B CCK-8 assay to detect the proliferation capacity of SW1990 cells. C-E Flow cytometry analysis of SW1990 cell death rates. F WST-8 colorimetric assay to measure NADPH levels in SW1990 cells. G Fluorescence microplate assay to determine cystine uptake capacity in SW1990 cells. H Representative immunofluorescence images of F-actin (phalloidin staining) and nuclei (DAPI) in SW1990 cells treated with RRx-001 or vehicle control. I Quantification of immunofluorescence staining. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human pancreatic ductal epithelial cells (HPDE-C7) and pancreatic cancer cells (SW1990) were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: CCK-8 Assay, Flow Cytometry, Colorimetric Assay, Fluorescence, Immunofluorescence, Staining, Control