摘要
目的:探讨上调miR-34b的表达对HCC827细胞对埃克替尼敏感性的影响及其机制。方法:应用细胞转染技术获得miR-34b高表达的HCC827细胞,并将HCC827细胞分为空白对照组、转染组、埃克替尼组及埃克替尼与转染联合组,采用MTT法测定各组细胞增殖的情况,运用流式细胞术分析各组细胞凋亡率及生长周期的变化,同时应用Real-time PCR及Western Blot方法检测各组c-MET基因mRNA及蛋白的表达水平。结果:miR-34b转染后,HCC827细胞的增殖能力下降,凋亡率升高(P<0.05);miR-34b转染后,埃克替尼对HCC827细胞的抑制率较埃克替尼组明显上升,半数抑制浓度(IC50)明显下降(P<0.05);转染组及埃克替尼与转染联合组的c-MET水平较对照组有所下降(P<0.05)。结论:上调miR-34b表达可以抑制HCC827细胞增殖,促进其凋亡,并可以增强HCC827细胞对盐酸埃克替尼的敏感性;其机制可能与miR-34b反馈抑制c-MET有关。
Objective： To discuss the effect of the increase of miR -34b on the sensitivity of HCC827 cells to Ico-tinib. Methods： According to the need,the experiment was divided into 4 groups： HCC827（control group） ,HCC827/ miR -34b group,HCC827/Icotinib,and HCC827/miR - 34b/Icotinib group. MTT assay and flow cytometry analysis were used to test the cell proliferation, apoptosis rate and cell cycle. Real - time PCR were used to detect c - MET mRNA levels. Western Blot was used to detect protein of c - MET. Results ： Transfection of miR - 34b can suppress the cell growth of HCC827 cells, and promote the apoptosis （ P 〈 0. 05 ） . Up - regulated miR - 34b expression in-creased the sensitivity of HCC827 cells to Icotinib,and the IC50 decreased apparently（P 〈0.05）. The expression of c -MET mRNA and protein in cells of HCC827/miR - 34b and HCC827/miR - 34b/Icotinib group were down - regu-lated significantly （P 〈0. 05）. Conclusion：These results suggested that miR -34b can suppress the cell growth andpromote the apoptosis. The higher expression of miR - 34b are associated with increased sensitivity of HCC827 cells to Icotinib. It may be related to the down - regulates of c - MET.
出处
《现代肿瘤医学》
CAS
2018年第5期676-680,共5页
Journal of Modern Oncology
