[关键词]
[摘要]
[摘要] 目的:探讨扭转原肠胚形成同系物1 基因(twisted gastrulation protein homolog 1,TWSG1)对胃癌MGC-803 细胞增殖及凋亡的影响。方法:设计3 条TWSG1 基因相关的siRNA,将对数生长期MGC-803 细胞分为空白对照组、阴性对照组、siRNA1干扰组、siRNA2 干扰组和siRNA3 干扰组,待细胞汇合率达70%~80%时分别转染空载体、连接siRNA1、siRNA2 和siRNA3 的载体。采用qPCR 和Western blotting 检测各组细胞TWSG1 mRNA和TWSG1 蛋白的表达水平,筛选干扰效率最高的稳定细胞株。用CCK-8 法检测各组细胞的增殖情况,用流式细胞术检测各组细胞的凋亡情况。结果:各转染组细胞中siRNA1 干扰效率最高。与空白对照组和阴性对照组相比,siRNA干扰组细胞TWSG1 表达下调(P<0.05),其细胞增殖显著增加(P<0.05),凋亡显著减少(P<0.05)。结论:siRNA干扰MGC-803 细胞中TWSG1基因的表达后,可促进细胞增殖、抑制细胞凋亡。
[Key word]
[Abstract]
[Abstract] Objective: To study the effects of twisted gastrulation protein homolog 1 (TWSG1) gene on proliferation and apoptosis of gastric cancer MGC-803 cells. Methods: Three siRNAs for TWSG1 gene were designed.The MGC-803 cells at logarithmic phase were divided into blank control group, negative control group (siRNA-NC), siRNA1 interference group, siRNA2 interference group and siRNA3 interference group by transfecting with relevant vectors. The mRNA and protein expressions of TWSG1 in each group were identified by qPCR and Western blotting, respectively; and the stable cell line with highest interference efficiency was screened.The proliferation of cells in each group was detected by CCK-8 assay, and the apoptosis of three groups was detected by flow cytometry. Results:The results of qPCR and Western blotting showed siRNA1 exhibited highest interference efficiency. Compared with the blank control group and the negative control group, the expression of TWSG1 in siRNA interference cell group was lower (P<0.05), the cell proliferation significantly increased (P<0.05), and apoptosis significantly reduced (P<0.05). Conclusion: siRNA interfering TWSG1 expression in MGC-803 cells can promote cell proliferation, inhibit cell apoptosis.
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[基金项目]
广东省科技计划资助项目(No.2015A010107005);广东省自然科学基金(No. 2017A030310022)