摘要
目的:研究Mig-6对肝癌细胞增殖和凋亡的影响,以探讨Mig-6在肝癌中的作用机制。方法:采用Mig-6过表达质粒和siRNA转染肝癌细胞,使用Real-time PCR和Western Blot法检测转染后的过表达或沉默效果;CCK-8实验检测细胞增殖水平的变化;流式细胞仪检测细胞凋亡百分比的变化;蛋白免疫印迹检测相关蛋白的表达变化。结果:转染Mig-6能抑制肝癌细胞的增殖,促进肝癌细胞的凋亡;干扰Mig-6能促进肝癌细胞的增殖,抑制肝癌细胞的凋亡。上调Mig-6能增加肝癌细胞Caspase-3的活性,抑制P-ERK的磷酸化;干扰下调Mig-6能抑制肝癌细胞Caspase-3的活性,增加P-ERK的磷酸化。结论:在肝癌细胞中,Mig-6表现出抑制细胞增殖及促进细胞凋亡的作用,该作用可能与Mig-6能增加Caspase-3的活性,同时抑制P-ERK的表达有关。
Objective: To investigate the effects of Mig- 6 gene over expression or silencing by plasmid or siRNA( small interfering RNA) interference on the proliferation and apoptotic of hepatocellular carcinoma( HCC),to identify the mechanism. Methods: Hep G- 2 and SMMC- 7721 were transfected with the plasmid or specific siRNA of Mig-6. The result of Mig- 6 silencing or over expression were examine by Real- time PCR. CCK- 8 assay was used to detect the cell proliferation. Flow cytometry and Annexin- V- FITC apoptosis assay were used to detect the apoptosis.Results: Transfection Mig- 6 plasmid can inhibit HCC cell proliferation,promote HCC apoptosis. Down regulation of Mig- 6 by siRNA can promote HCC cell proliferation and inhibit cell apoptosis. Over expression of Mig- 6 can increase the activity of Caspase- 3,inhibit ERK phosphorylation in HCC cell. Interference Mig- 6 in HCC can inhibit Caspase- 3 activity and increase the P- ERK. Conclusion: In HCC cells,Mig- 6 can inhibit cell proliferation,promote cell apoptosis,this effect may be related to Mig- 6 increasing the activity of Caspase- 3,inhibit the expression of P- ERK.
出处
《现代肿瘤医学》
CAS
2016年第12期1857-1861,共5页
Journal of Modern Oncology
基金
国家自然科学基金资助项目(编号:81501564)
