The effects of short interfering RNAs on E2F-1 gene expression and biological behaviors of gastric cancer ceIl line MGC803
Yongshuo Yin, Lei Li, Yubo Xie
Abstract
Yongshuo Yin, Lei Li, Yubo Xie
Abstract
Objective To investigate the effects of RNA interference(RNAi)targeting E2F-1 on biological behaviors of gastric cancer cell line MGC803.Methods E2F-1-siRNA or negative control-siRNA was transfected into gastric cancer cell line MGC803 cells.Forty-eight h later,the expression of E2F-1 mRNA and protein in gastric cancer cell fine MGC803 was detected by real-time quantitative PCR and Western blot respectivelv.Cell cycle was examined by flow cytometry,and the proliferative rate of MGC803 cells tegted by MTT assay.Results E2F-1-siRNA effectively inhibited the expression of E2F-1 mRNA and Drotein and influenced proliferation of MGC803 cells.As compared with negative control and nontmsfected groups,the expression levels of E2F-1 mRNA and protein were decreased by 84.8%,85.3%(P<0.05),and 77.2%,79.6%,and proliferation of MGC803 cells was suppressed by 69.0%and 81.6%(P<0.05)in E2F-1-siRNA transfection group,respectively.E2F-1-siRNA promoted accumulation of more DNA of MGC803 cells at G2/M phase,and the rate of DNA content in G2/M phase in E2F-1-siRNA trailsfection group was(54.98±3.46)%,which was significantly higher than(43.07±3.82)% in negative control group,and(31.47±2.12)%in non-transfection group(P<0.05).Conclusion E2F-1 gene repression by RNAi can decrease the DNA content in G1 phase of MGC803 cells,which can arrest the cells in G2/M phase and inhibit the proliferation of MGC803 cells. Key words: Gastric carcinoma; Small interfering RNA; Cell cycle; Proliferation
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Objective To investigate the effects of RNA interference(RNAi)targeting E2F-1 on biological behaviors of gastric cancer cell line MGC803.Methods E2F-1-siRNA or negative control-siRNA was transfected into gastric cancer cell line MGC803 cells.Forty-eight h later,the expression of E2F-1 mRNA and protein in gastric cancer cell fine MGC803 was detected by real-time quantitative PCR and Western blot respectivelv.Cell cycle was examined by flow cytometry,and the proliferative rate of MGC803 cells tegted by MTT assay.Results E2F-1-siRNA effectively inhibited the expression of E2F-1 mRNA and Drotein and influenced proliferation of MGC803 cells.As compared with negative control and nontmsfected groups,the expression levels of E2F-1 mRNA and protein were decreased by 84.8%,85.3%(P<0.05),and 77.2%,79.6%,and proliferation of MGC803 cells was suppressed by 69.0%and 81.6%(P<0.05)in E2F-1-siRNA transfection group,respectively.E2F-1-siRNA promoted accumulation of more DNA of MGC803 cells at G2/M phase,and the rate of DNA content in G2/M phase in E2F-1-siRNA trailsfection group was(54.98±3.46)%,which was significantly higher than(43.07±3.82)% in negative control group,and(31.47±2.12)%in non-transfection group(P<0.05).Conclusion E2F-1 gene repression by RNAi can decrease the DNA content in G1 phase of MGC803 cells,which can arrest the cells in G2/M phase and inhibit the proliferation of MGC803 cells. Key words: Gastric carcinoma; Small interfering RNA; Cell cycle; Proliferation
Key concepts: Transfection, Molecular biology, E2F, Cell cycle, Small interfering RNA, Cell growth, RNA interference, Cell culture