摘要
目的研究沉默信息调节因子1(SIRT1)在胃癌细胞增殖和凋亡中的作用。方法构建包装针对SIRT1的shRNA慢病毒载体,感染胃癌细胞及胃黏膜上皮细胞;应用荧光显微镜观察并应用免疫印迹方法证实转染的可靠性;应用CCK-8法检测shRNA-SIRT1对胃癌细胞及胃黏膜上皮细胞增殖的影响;APC-Annexin V标记法,流式细胞仪检测shRNA-SIRT1对胃癌细胞及胃黏膜上皮细胞凋亡的影响。结果成功进行了携带以SIRT1为靶向的shRNA的慢病毒包装,并成功转染到人胃癌细胞中。通过对比研究转染shRNA-SIRT1和空转对照组的胃癌细胞,发现shRNA可以通过沉寂胃癌细胞中的SIRT1从而抑制了胃癌细胞的增殖,促进其凋亡,而对胃黏膜上皮细胞的增殖凋亡影响较小。结论通过靶向应用于SIRT1的shRNA技术抑制了胃癌细胞的增殖,促进其凋亡,进一步明确SIRT1在胃癌细胞增殖和发展的作用,而且为生物基因治疗胃癌提供了实验基础。
Objective To study the effect of SIRT1 in gastric cancer cell proliferation and apoptosis. Methods Lentivirus expressing SIRT1-target shRNA was produced by plasmid. Gastric cancer cell lines and gastric epithelial cells were infected by the recombinant lentivirus. Fluorescence microscope and immunoblotting were used to confirm the validation of lentivirus transfection. CCK-8 analysis was used to test the effect of shRNA-SIRT1 on proliferation of gastric cancer cells and gastric epithelial cells;APCAnnexin V analysis and flow cytometry were used to test the effect of shRNA-SIRT1 on apoptosis of gastric cancer cells and gastric epithelial cells. Results Lentivirus expressing SIRT1-target shRNA was produced successfully by plasmid. Gastric cancer cell lines and gastric epithelial cells were successfully infected by lentivirus. The shRNA can suppress the proliferation of gastric cancer cells and promote the apoptosis by silencing SIRT1,nearly no effect on gastric epithelial cells. Conclusion Application of SIRT1-target shRNA can suppress proliferation and promote apoptosis of gastric cancer cell. We conclude that SIRT1 plays a role in gastric cell proliferation and apoptosis, thus providing a potential choice for the development of novel biological gene therapy.
出处
《中华保健医学杂志》
2014年第2期117-120,共4页
Chinese Journal of Health Care and Medicine
关键词
SIRT1
胃癌
细胞增殖
凋亡
SIRT1
Gastric cancer
Cell proliferation
Apoptosis
