Journal: Cancer Medicine

Article Title: CCNG1 (Cyclin G1) regulation by mutant‐P53 via induction of Notch3 expression promotes high‐grade serous ovarian cancer (HGSOC) tumorigenesis and progression

doi: 10.1002/cam4.1812

Figure Lengend Snippet: Effects of down‐regulating CCNG1 on the migration and invasion ability of ovarian cancer cell lines. The number of migrating and invasion cells in silenced CCNG1group was significantly reduced in A2780 (A) or HO8910 (B). The numerical values were mean ± SD of three replicates. All the experiments were repeated three times using the same batch of cells

Article Snippet: Human ovarian cancer cell line A2780 (Procell, China, CL‐0013) and HO8910 (Procell, China, CL‐0113) were cultured in RPMI 1640 culture medium and SKOV3 (ATCC, USA, HBT‐77) in McCoy's 5A medium, supplemented with 10% fetal bovine serum (FBS, Gibco).

Techniques: Migration

Journal: Cancer Medicine

Article Title: CCNG1 (Cyclin G1) regulation by mutant‐P53 via induction of Notch3 expression promotes high‐grade serous ovarian cancer (HGSOC) tumorigenesis and progression

doi: 10.1002/cam4.1812

Figure Lengend Snippet: CCNG1 modulated cisplatin sensitivity. A, After exposure to cisplatin, CCNG1 protein expression in ovarian cancer cells was time‐dependently up‐regulated. B and C, Cell inhibitory rates and IC50 of cisplatin in HO8910 and A2780 were significantly changed after down‐regulation of CCNG1 expression by shRNA. Cell viability detection, from cells seeded in 96‐well plates to calculate IC50, was repeated three times to obtain three IC50

Article Snippet: Human ovarian cancer cell line A2780 (Procell, China, CL‐0013) and HO8910 (Procell, China, CL‐0113) were cultured in RPMI 1640 culture medium and SKOV3 (ATCC, USA, HBT‐77) in McCoy's 5A medium, supplemented with 10% fetal bovine serum (FBS, Gibco).

Techniques: Expressing, shRNA

Journal: Cancer Medicine

Article Title: CCNG1 (Cyclin G1) regulation by mutant‐P53 via induction of Notch3 expression promotes high‐grade serous ovarian cancer (HGSOC) tumorigenesis and progression

doi: 10.1002/cam4.1812

Figure Lengend Snippet: Down‐regulation of CCNG1 reduced tumor metastasis in vivo . A, H&E staining of lung metastasis of shCCNG1‐A2780 cell line and its control. B, Nude mice injected with shCCNG1‐A2780 cell line developed fewer lung metastatic foci than its control ( P < 0.0001). C, CCNG1 was regulated by P53mt via induction of Notch3 expression, which ultimately promotes both HGSOC tumor cell metastasis and cisplatin resistance

Article Snippet: Human ovarian cancer cell line A2780 (Procell, China, CL‐0013) and HO8910 (Procell, China, CL‐0113) were cultured in RPMI 1640 culture medium and SKOV3 (ATCC, USA, HBT‐77) in McCoy's 5A medium, supplemented with 10% fetal bovine serum (FBS, Gibco).

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

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin inhibits the proliferation of ovarian cancer cells. EdU assays demonstrate that metformin reduces proliferation of (A) SKOV3, (B) A2780 and (C) ES2 cells. Representative fluorescence microscopy images of EdU incorporation and Hoechst 33342 staining of nuclei (magnification, ×100). The proliferation rates of SKOV3, A2780, and ES2 cells were calculated. Data are presented as the mean ± standard deviation. * P<0.05; ** P<0.01; *** P<0.001. Met, metformin; Glu, glucose; EdU, 5-ethynyl-20-deoxyuridine; NC, negative control.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Fluorescence, Microscopy, Staining, Standard Deviation, Negative Control

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin promotes the apoptosis of ovarian cancer cells. Percentage of apoptotic (A) SKOV3, (B) A2780 and (C) ES2 cells were detected by flow cytometry analysis using Annexin V-fluorescein isothiocyanate/PI staining. Apoptotic cells includes Annexin V(-)/PI(+), Annexin V(+)/PI(−) and Annexin V(+)/PI(+) cells. Data are presented the mean ± standard deviation. * P<0.05; ** P<0.01. PI, propidium iodide; Met, metformin; Glu, glucose; NC, negative control.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Flow Cytometry, Staining, Standard Deviation, Negative Control

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin inhibits migration of ovarian cancer cells. Migration of (A) SKOV3, (B) A2780 and (C) ES2 cells were measured by scratch wound healing assay. Representative bright-field images (magnification, ×100) and quantification of recovered distance at 24 h after 10 mM metformin treatment. Data are presented the mean ± standard deviation. * P<0.05; ** P<0.01; *** P<0.001. Met, metformin; Glu, glucose; NC, negative control.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Migration, Wound Healing Assay, Standard Deviation, Negative Control

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin inhibits migration of ovarian cancer cells. Migration of (A) SKOV3, (B) A2780 and (C) ES2 cells were measured by Transwell migration assay. Representative bright-field images of the cell migration (magnification, ×100) and quantification of migrated cells at 24 h after 10 mM metformin treatment are shown. Data are presented the mean ± standard deviation. * P<0.05; ** P<0.01; *** P<0.001. Met, metformin; Glu, glucose; NC, negative control.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Migration, Transwell Migration Assay, Standard Deviation, Negative Control

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin decreases H3K27me3 and expression of polycomb repressor complex 2 in ovarian cancer cells. The protein expression of (A) H3K27me3 and (B) EZH2, SUZ12 and EED in A2780, SKOV3 and ES2 cells were assayed by western blot analysis. Met, metformin; Glu, glucose; H3K27me3, histone H3 lysine 27 trimethylated; H3, histone H3; EZH2, histone-lysine N-methyltransferase EZH2; SUZ12, polycomb protein SUZ12; EED, polycomb protein EED.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Expressing, Western Blot

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin decreases PRC2 of ovarian cancer cells. mRNA expression of EZH2, SUZ12 and EED in (A) SKOV3, (B) A2780 and (C) ES2 cells determined by reverse transcription-quantitative polymerase chain reaction analysis. Data are presented the mean ± standard deviation. * P<0.05; ** P<0.01; *** P<0.001. Met, metformin; Glu, glucose; EZH2, histone-lysine N-methyltransferase EZH2; SUZ12, polycomb protein SUZ12; EED, polycomb protein EED.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: International Journal of Oncology

Article Title: Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation

doi: 10.3892/ijo.2018.4343

Figure Lengend Snippet: Metformin inhibits H3K27me3 through the AMPK pathway. (A) Metformin activates AMPK of ovarian cancer cells in normoglycemic condition. (B) 2-DG activates AMPK and suppresses H3K27me3, PRC2 of ovarian cancer cells in normoglycemic condition. (C) Compound C inhibits the effect of metformin on PRC2 and H3K27me3 when ovarian cancer cells were cultured in normoglycemic condition. The protein expression of p-AMPKα, AMPKα, H3K27me3, EZH2, SUZ12 and EED in A2780, SKOV3 and ES2 cells were assayed by western blot analysis. The expression was normalized to β-actin. Met, metformin; Glu, glucose; p-AMPKα, phospho-AMPKα; AMPKα, AMP-activated protein kinase; 2-DG, 2-deoxy-D-glucose; EZH2, histone-lysine N-methyltransferase EZH2;SUZ12, polycomb protein SUZ12; EED, polycomb protein EED; 2-DG, dorsomorphin 2HCl; H3K27me3, histone H3 lysine 27 trimethylation.

Article Snippet: Human epithelial ovarian cancer cell lines SKOV3 (ovarian adenocarcinoma), ES2 (ovarian clear cell carcinoma), and A2780 (ovarian carcinoma) were purchased from China Center for Type Culture Collection (Wuhan University, Wuhan, China) and were cultured in Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) containing 25 mM glucose (mimicking hyperglycemia) or 5.5 mM glucose (mimicking normoglycemia) supplemented with 10% fetal bovine serum (v/v; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China) at 37°C in 95% air and 5% CO 2 .

Techniques: Cell Culture, Expressing, Western Blot

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: miR-1299 is downregulated in OC and acts as a negative regulator of NOTCH3. (A) Analysis of miR-1299 expression in OC and normal ovary tissues (NC) using RT-qPCR. (B) miR-1299 expression was significantly lower in OC tissues with high NOTCH3 expression compared to those with low NOTCH3 expression. (C) Correlation between miR-1299 and NOTCH3 levels in OC cell lines. (D) RT-qPCR analysis of NOTCH3 pathway genes after transfection of miR-1299 mimics in A2780 cells. (E) Western blot analysis of NOTCH3 after transfection of miR-1299 mimics in A2780 and CAOV3 cell lines. (F) Putative miR-1299 binding sequence in NOTCH3 3′UTR (untranslated region) (WT) and the mutated 3′UTR sequence (MUT). (G) Relative luciferase activity of reporter plasmids carrying wild-type (WT) or mutant (MUT) NOTCH3 3′UTR in A2780 cells co-transfected with NC or miR-1299 mimics. Means ± SD are shown. Statistical analysis was conducted using Student's t-test. *P<0.05 and **P<0.01; ns, not significant. OC, ovarian cancer; NOTCH3, notch receptor 3; HES1, hairy and enhancer of split-1; HEY1, Hes related family BHLH transcription factor with YRPW motif 1; HEY 2, Hes related family BHLH transcription factor with YRPW motif 2; JAG1, Jagged canonical Notch ligand 1; PBX1, Pre-B-cell leukemia transcription factor 1.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Binding Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: Upregulation of miR-1299 inhibits cell proliferation, colony formation, EdU incorporation, and cell cycle in OC A2780 and CAOV3 cell lines. (A) RT-qPCR analysis of miR-1299 expression level in OC cells after transfection of NC or miR-1299 mimics for 48 h. (B) Cell proliferation of OC cells after transfection of NC or miR-1299 mimics by CCK-8 assay. (C) Representative photographs and quantifications of the colony formation assay after transfection of NC or miR-1299 mimics. (D) Representative photographs of EdU incorporation assay in OC cells after transfection of NC or miR-1299 mimics. (E) Cell cycle distribution of OC cells after transfection of NC or miR-1299 mimics by flow cytometry. Means ± SD are shown. Statistical analysis was conducted using Student's t-test. *P<0.05 and **P<0.01, miR-1299 mimic compared with the NC mimic; ns, not significant; OC, ovarian cancer.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: lncRNA TUG1 functions as a sponge of miR-1299 and promotes cell proliferation in OC. (A) Screening methods for regulatory lncRNAs of miR-1299 in OC. (B) The two highest scoring putative miR-1299 binding sites in the TUG1 sequence (WT) and the point mutations in either binding site (MUT). (C) Relative luciferase activity of reporter plasmids carrying wild-type (WT) or mutant (MUT) TUG1 binding sites in A2780 cells co-transfected with NC or miR-1299 mimics. (D) Confirmation of TUG1 knockdown in A2780 cells transfected with TUG1 shRNA by RT-qPCR analysis. (E) RT-qPCR analysis of miR-1299 level in A2780 cells transfected with TUG1 shRNA or scrambled shRNA. Results of the (F) cell proliferation, (G) colony formation, (H) EdU incorporation assays, and (I) cell cycle analysis of A2780 cells transfected with NC-sh, TUG1-sh1, TUG1-sh2, and TUG1-sh+miR-1299 inhibitors. (J) Representative blot image of NOTCH3 protein level in A2780 cells transfected with NC-sh, TUG1-sh1, TUG1-sh2, and TUG1-sh+miR-1299 inhibitors by western blotting. Means ± SD are shown. Statistical analysis was conducted using Student's t-test and one-way ANOVA with Tukey post hoc test. *P<0.05 and **P<0.01; ns, not significant; OC, ovarian cancer; NOTCH3, notch receptor 3; TUG1, lncRNA taurine upregulated gene 1.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Binding Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis, Transfection, Knockdown, shRNA, Quantitative RT-PCR, Cell Cycle Assay, Western Blot

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: TUG1 is a potential downstream target of NOTCH3. (A) Schematic diagram showing RBP Jκ motifs around the transcriptional start site (TSS) of the TUG1 gene. (B and C) TUG1 expression was inversely correlated with NOTCH3 mRNA in fresh OC tissues (B) and in the TCGA database (C). (D) RT-qPCR analysis of TUG1 level in A2780 cells treated with DAPT. (E) Diagrams showing the miR-1299/NOTCH3/TUG1 feedback loop in the development of OC. Statistical analysis was conducted using one-way ANOVA with Tukey post hoc test and Pearson's correlation analysis. **P<0.01; ns, not significant; OC, ovarian cancer; NOTCH3, notch receptor 3; TUG1, lncRNA taurine upregulated gene 1; NICD3, NOTCH3 intracellular domain; HES1, hairy and enhancer of split-1; HEY1, Hes related family BHLH transcription factor with YRPW motif 1; HEY 2, Hes related family BHLH transcription factor with YRPW motif 2.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR