Effects of RNAi-mediated inhibitor of growth 1 gene silencing on apoptosis of gastric cancer cell line AGS
Xiangmin He
Abstract
Xiangmin He
Abstract
Objective To explore the effects of RNAi-mediated inhibitor of growth 1 (ING1)gene silencing on cell apoptosis of human gastric cancer cell line AGS.Methods After cultured,human gastric cell line AGS was divided into blank control group, negative control group (transfected with negative control siRNA sequence) and siRNA group (transfected with specific ING1 siRNA sequence).After transfection, the ING1 gene silencing effects at different time points were detected by immunofluorescence, real-time PCR and Western blot.The effects of ING1 gene silencing on apoptosis of AGS cells were evaluated by flow cytometry.Resnlts ING1 expression was restricted to the AGS cytoplasm.In real time PCR testing, the ING1 gene expression in blank control group was set as 1.24 h and 40 h after transfection, the ING1 gene relative expression quantity of negative control group was 0.88±0.16 and 0.92±0.13, and that of siRNA group was 0.38±0.09 and 0.17±0.06,respectively.Compared blank control group with negative control group, P values were 0.78 and 0.82.Compared blank control group with siRNA group, both P values were 0.01.Compared negative control group with siRNA group, P value was 0.02 and 0.01.In western blot testing, the protein expression of ING1 decreased significantly after transfected for 40 h.The apoptosis rate of blank control group was 11.06% ±0.97%, and 40 h after transfection, the apoptosis rate in AGS cells of negative control group and siRNA group was 11.82 % ± 0.69 % and 6.70% ± 0.41%, respectively.Compared negative control group with siRNA group, the P value was 0.024.Compared blank control group with negative control group, the P value was 0.76.Compared negative control group with siRNA group, the P value was 0.019.Conclusion ING1 plays an important role in cell apoptosis of human gastric adenocarcinoma cells, and may become a new target for gastric cancer gene therapy. Key words: RNA interference; Stomach neoplasms; Adenocarcinoma; Genes, Tumor suppressor; Gene expression; Apoptosis
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Objective To explore the effects of RNAi-mediated inhibitor of growth 1 (ING1)gene silencing on cell apoptosis of human gastric cancer cell line AGS.Methods After cultured,human gastric cell line AGS was divided into blank control group, negative control group (transfected with negative control siRNA sequence) and siRNA group (transfected with specific ING1 siRNA sequence).After transfection, the ING1 gene silencing effects at different time points were detected by immunofluorescence, real-time PCR and Western blot.The effects of ING1 gene silencing on apoptosis of AGS cells were evaluated by flow cytometry.Resnlts ING1 expression was restricted to the AGS cytoplasm.In real time PCR testing, the ING1 gene expression in blank control group was set as 1.24 h and 40 h after transfection, the ING1 gene relative expression quantity of negative control group was 0.88±0.16 and 0.92±0.13, and that of siRNA group was 0.38±0.09 and 0.17±0.06,respectively.Compared blank control group with negative control group, P values were 0.78 and 0.82.Compared blank control group with siRNA group, both P values were 0.01.Compared negative control group with siRNA group, P value was 0.02 and 0.01.In western blot testing, the protein expression of ING1 decreased significantly after transfected for 40 h.The apoptosis rate of blank control group was 11.06% ±0.97%, and 40 h after transfection, the apoptosis rate in AGS cells of negative control group and siRNA group was 11.82 % ± 0.69 % and 6.70% ± 0.41%, respectively.Compared negative control group with siRNA group, the P value was 0.024.Compared blank control group with negative control group, the P value was 0.76.Compared negative control group with siRNA group, the P value was 0.019.Conclusion ING1 plays an important role in cell apoptosis of human gastric adenocarcinoma cells, and may become a new target for gastric cancer gene therapy. Key words: RNA interference; Stomach neoplasms; Adenocarcinoma; Genes, Tumor suppressor; Gene expression; Apoptosis
Key concepts: Gene silencing, Transfection, Molecular biology, Apoptosis, RNA interference, Biology, Cell culture, Gene expression