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[摘要]
摘要 目的:探究miR-365对胃癌细胞的作用机制。方法:提取人正常胃黏膜上皮细胞RGM-1和胃癌SGC-7901内的miR-365,通过RT-PCR测定其表达的差异;采用脂质体转染的方法将miR-365转入细胞中。MTT法检测转染后的SGC-7901增殖情况;流式细胞术检测转染后细胞的凋亡情况;Western blotting检测转染48h后p21及Cyclin D1的表达。结果: miR-365的表达显著低于RGM-1细胞 (P<0.05)。转染miR-365 mimics后, miR-365转染组中miR-365表达显著高于空白对照组和阴性对照组。转染2d后,空白对照组中miR-365的OD值显著高于干扰组细胞,差异显著(P<0.05)。miR-365 转染组细胞的凋亡率明显增加 (P <0.05)。转染后的p21蛋白表达量显著升高,而Cyclin D1的表达量显著下降 (P <0.05).结论:转染miR-365促进细胞凋亡的同时也抑制了胃癌细胞的增殖。
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[Abstract]
Objective: To investigate the effect of microRNA-365 on the proliferation of gastric cancer and its mechanism. Methods: Real-time fluorescence quantitative PCR was used to detect the difference of the expression of microRNA-365 in human normal gastric mucosal epithelial cells RGM-1 and gastric cancer SGC-7901 cells; Liposome transfection was used to change the expression of microRNA-365 in cells; MTT was used to detect the proliferation of transfected SGC-7901 cells; flow cytometry was used to detect the apoptosis of SGC-7901 cells after transfected; Western blotting was used to detect the expression of p21 and cyclin D1 protein after 48 hours of transfection. Results: The expression of microRNA-365 was significantly lower than that epithelial RGM-1 cells (P < 0.05). the expression of microRNA-365 in the microRNA-365 transfection group increased significantly (P < 0.05). the OD value of the cells in the blank control group was significantly higher than that in the interference group 2 days after transfection (P < 0.05); The apoptotic rate of microRNA-365 transfection group increased significantly (P < 0.05). After transfection, the expression level of p21 protein increased significantly , and the relative expression level of Cyclin D1 decreased significantly (P < 0.05). Conclusion: Transfection of microRNA-365 can promote apoptosis and inhibit the proliferation of gastric cancer cells.
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