miRNA-34 a对人结肠癌细胞 HCT116增殖及侵袭转移的影响

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。

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

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-34a eukaryotic expression vector was designed and established to stably transfect HCT116 cells.Realtime PCR was used to verify the expression of miR-34a after expression vector transfection.MTT assay was used to determine the changes of HCT116 cell prolifera-tion.Transwell assay was performed to measure the cell invasion ability of HCT116 cells.Results After transfection of miR-34a into HCT116 cells,the expression of miR-34a increased by(7.32 ±1.34) folds and cell proliferation ability was significantly in-hibited,which was reduced to(0.49 ±0.10) compared with the control group.Bcl-2 protein expression was inhibited by miR-34a transfection,but the expression of BAX was enhanced by miR-34a transfection.The invasion of HCT116 cells was also significant-ly increased by miR-34a 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-34a can inhibit the prolifera-tion and invasion in human colon cancer cells HCT116,and it is closely correlated with the regulation of Bcl-2/BAX and the ex-pression of MMP-2 and-9 proteins.