摘要
目的:研究胃癌细胞SGC7901中RUNX2的表达,以及siRNA沉默RUNX2的表达后,对人胃癌细胞SGC7901的RUNX2的表达、细胞增殖和凋亡的影响.方法:将细胞分为以下3组:空白对照组、空载体组、实验组(即转染RUNX2siRNA组).用针对RUNX2特异靶点的siRNA转染胃癌细胞SGC7901,运用RT-PCR、Westernblot检测RUNX2mRNA和蛋白水平的变化;MTT检测细胞增殖的影响;流式细胞仪检测细胞凋亡的变化.所有实验均重复3次.结果:在胃癌细胞SGC7901中存在RUNX2的表达,siRNA沉默RUNX2的表达后,与空白对照组相比,RUNX2在mRNA(0.27±0.068vs0.45±0.058,F=75.6,P<0.01)和蛋白水平(F=123.8,P<0.001)的表达均下降,差异有显著统计学意义.siRNA转染24、48、72h,MTT检测胃癌细胞SGC7901的增殖率与空白对照组相比明显下降(0.23±0.039vs0.32±0.012;0.31±0.037vs0.45±0.074;0.52±0.021vs0.72±0.006;F=173.744、14.012、253.145;均P<0.001),流式细胞术检测RUNX2siRNA组72h细胞凋亡明显高于空白对照组及空载体组(45.65±0.64vs4.46±0.27,4.23±0.33,均P<0.01).结论:胃癌细胞SGC7901中存在RUNX2的表达,siRNA抑制RUNX2的表达可抑制胃癌细胞的增殖,促进凋亡,RUNX2可望成为胃癌基因治疗的新的靶点.
AIM：To detect the expression of runt-related transcription factor gene 2（RUNX2） in human gastric cancer cell line SGC7901 and to investigate the influence of small interfering RNA（siRNA）-mediated silencing of the RUNX2 gene on the proliferation and apoptosis of SGC7901 cells.METHODS：SGC7901 cells were divided into three groups：blank control group,negative control group（transfected with an empty vector）,and experiment group（transfected with RUNX2 siRNA）.After SGC7901 cells were transfected with RUNX2 siRNA,the mRNA and protein expression of RUNX2 was examined by RT-PCR and Western blot,respectively;cell proliferation was evaluated by MTT assay;and cell apoptosis was detected by ？ ow cytometry（FCM）.RESULTS：Compared with cells of the blank control group,the expression of RUNX2 mRNA（0.27 ± 0.068 vs 0.45 ± 0.058,F = 75.6,P 0.01） and protein（F = 123.8,P 0.001） was down-regulated in cells transfected with RUNX2 siRNA.At 24,48,and 72 h after transfection,the proliferation rates of SGC7901 cells transfected with RUNX2 siRNA were significantly lower than those of non-transfected cells（0.23 ± 0.039 vs 0.32 ± 0.012;0.31 ± 0.037 vs 0.45 ± 0.074;0.52 ± 0.021 vs 0.72 ± 0.006;F = 173.744,14.012,253.145;all P 0.001）.The apoptosis rate of SGC7901 cells transfected with RUNX2 siRNA was signif icantly higher than those of cells of the blank control group and negative control（45.65 ± 0.64 vs 4.46 ± 0.27,4.23 ± 0.33,both P 0.01）.CONCLUSION：RUNX2 expression was detected in SGC7901 cells.SiRNA-mediated silencing of the RUNX2 gene can inhibit proliferation and induce apoptosis in SGC7901 cells.RUNX2 may be a new gene therapy target for gastric cancer.
出处
《世界华人消化杂志》
CAS
北大核心
2011年第4期338-343,共6页
World Chinese Journal of Digestology
关键词
胃癌
人类相关转录基因2
小分子干扰核苷酸
凋亡
Gastric cancer
Runt-related transcription factor gene 2
Small interfering RNA
Apoptosis
