miRNA-34a对人结肠癌细胞HCT116增殖及侵袭转移的影响

Effects of miRNA-34a on Cell Proliferation and Invasion in Human Colon Cancer Cell HCT116

目的研究微小RNA-34a(microRNA-34a,miR-34a)在人结肠癌细胞株HCT116中的过表达对细胞增殖和侵袭的影响及其机制。方法设计合成并构建携带绿色荧光蛋白(GFP)的miR-34a真核表达载体,脂质体法稳定转染HCT116细胞。通过Realtime PCR验证转染后miR-34a的表达变化。MTT法检测转染前后HCT116细胞增殖能力变化,Transwell法检测转染前后HCT116细胞侵袭能力改变,western blot检测目的蛋白的表达。结果 miR-34a转染HCT116细胞后其表达量增加到(7.32±1.34)倍,而HCT116细胞增殖能力也受到显著抑制,下降到(0.49±0.10);Bcl-2蛋白受到miR-34a表达的抑制,而BAX的表达则受到miR-34a表达的增强;HCT116细胞侵袭能力在miR-34a过表达后显著增强,相应的MMP-2和MMP-9表达也出现显著增强。阴性对照组与空白对照组比较无显著性差异。结论 miR-34a的过表达能够抑制人结肠癌细胞HCT116的增殖及侵袭转移,与调控Bcl-2/BAX和MMP-2和-9蛋白的表达密切相关。

Objective To investigate the effects of microRNA-34a（ miR-34a） overexpression on cell proliferation and invasion in human colon cancer cell line HCT116. Methods A portable green synthesis fluorescent protein（ GFP） of miR-34 a eukaryotic expression vector was designed and established to stably transfect HCT116 cells. Realtime PCR was used to verify the expression of miR-34 a after expression vector transfection. MTT assay was used to determine the changes of HCT116 cell proliferation. Transwell assay was performed to measure the cell invasion ability of HCT116 cells. Results After transfection of miR-34 a into HCT116 cells,the expression of miR-34 a increased by（ 7. 32 ± 1. 34） folds and cell proliferation ability was significantly inhibited,which was reduced to（ 0. 49 ± 0. 10） compared with the control group. Bcl-2 protein expression was inhibited by miR-34 a transfection,but the expression of BAX was enhanced by miR-34 a transfection. The invasion of HCT116 cells was also significantly increased by miR-34 a transfection,which was accompanied by MMP-2 and MMP-9 up-regulation. There was no significant difference in the negative control group and the control group. Conclusion Overexpression of miR-34 a can inhibit the proliferation and invasion in human colon cancer cells HCT116,and it is closely correlated with the regulation of Bcl-2 / BAX and the expression of MMP-2 and-9 proteins.