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目的探讨沉默磷脂酰肌醇3-激酶(PI3K)p110β因子的表达对人胃癌细胞生长的影响。方法运用RNA干扰技术,合成靶向抑制PI3Kp110β的小干扰RNA(si RNA)序列并分别瞬时转染人胃癌细胞株AGS、SGC7901和MKN45(si-PI3Kp110β),作为实验组,未转染的细胞为空白对照组(Con),转染无义序列的si RNA为阴性对照组(si-NC)。采用RT-PCR和Western Blot方法验证瞬时转染后PI3Kp110β mRNA和蛋白在空白对照组(Con)、阴性对照组(si-NC)和实验组(si-PI3Kp110β)细胞中的表达水平,并通过MTT法检测细胞增殖及Annexin V-FITC/PI双染法检测AGS细胞凋亡率的情况。结果小分子干扰RNA(si RNA)瞬时转染人胃癌细胞株AGS、SGC7901和MKN45后,能有效下调PI3Kp110β在细胞内的表达水平。与阴性对照组(si-NC)比较,静默PI3Kp110β因子可减少AGS细胞的增殖(P<0.05),并明显增加AGS细胞的凋亡率(P<0.01)。结论小分子干扰RNA可有效下调PI3Kp在人胃癌细胞中的表达,沉默PI3Kp110β因子可有效抑制胃癌肿瘤细胞的增殖,促进细胞凋亡,提示针对PI3Kp110β蛋白的干预治疗有可能成为抑制胃癌细胞生长的一个新的治疗方法。
Objective To investigate the effect of silencing the expression of PI3K (p110β) on the growth of human gastric cancer cells. Methods RNAi was used to synthesize small interfering RNA (siRNA) sequences targeting PI3Kp110β and transiently transfected into human gastric cancer cell lines AGS, SGC7901 and MKN45 (si-PI3Kp110β) respectively. The untransfected cells were Blank control group (Con), transfected with nonsense sequence of si RNA negative control group (si-NC). The expression of PI3Kp110β mRNA and protein in the blank control group (si), control group (si-NC) and experimental group (si-PI3Kp110β) after transfection was verified by RT-PCR and Western Blot. Method to detect cell proliferation and Annexin V-FITC / PI double staining method to detect the apoptosis rate of AGS cells. Results Transient siRNA transfected human gastric cancer cell lines AGS, SGC7901 and MKN45 could effectively down-regulate the expression of PI3Kp110β in the cells. Compared with negative control group (si-NC), silencing PI3Kp110βprotein could reduce the proliferation of AGS cells (P <0.05) and significantly increase the apoptosis rate of AGS cells (P <0.01). Conclusions Small interfering RNA (siRNA) can effectively down-regulate the expression of PI3Kp in human gastric cancer cells. Silencing PI3Kp110β can effectively inhibit the proliferation of gastric cancer cells and promote the apoptosis of gastric cancer cells. It suggests that the intervention of PI3Kp110β may be one of the mechanisms that inhibit the growth of gastric cancer cells New treatment.