HMGA2基因沉默对结肠癌LS 174T细胞增殖和侵袭的影响

Effects of HMGA2 RNA interference on proliferation and invasion of colon cancer LS 174T cells

目的探讨利用小发夹RNA(short hairpin RNA,shRNA)干扰技术沉默高迁移率族蛋白A2(high mobility group protein AT-hook 2,HMGA2)基因,观察对结肠癌LS 174T细胞增殖和侵袭的影响。方法构建HMGA2基因特异性shRNA慢病毒载体,干扰结肠癌LS 174T细胞,设立空白对照组(MOCK)、阴性对照组(Scramble shRNA)和HMGA2 shRNA三组,利用Real-time PCR和Western blot方法分别从基因和蛋白水平检测shRNA对HMGA2基因的干扰效果;MTT法检测抑制HMGA2基因表达后LS 174T细胞增殖情况;Transwell侵袭实验检测HMGA2基因沉默后LS 174T细胞侵袭能力的变化。结果转染shHMGA2慢病毒后,结肠癌LS 174T细胞中HMGA2 mRNA和蛋白表达均明显受到抑制(P<0.01);MTT法检测各组细胞的增殖能力,shHMGA2组细胞增殖水平明显低于空白对照组和阴性对照组(P<0.05);敲除HMGA2基因可明显降低LS 174T细胞的侵袭能力,空白对照组和阴性对照组平均穿膜细胞数分别为(124.28±65)/视野和(118.52±15)/视野,shHMGA2组平均穿膜细胞数为(53.35±8.45)/视野(P<0.05)。结论 shHMGA2慢病毒能明显下调HMGA2基因的表达,在体外可有效抑制结肠癌LS 174T细胞的增殖和侵袭。

Objective To investigate the effect of HMGA2 gene short hairpin RNA（shRNA） on proliferation and invasion of human colon cancer LS 174T cell line.Methods The recombinant lentivirus vector containing ShRNA targeting HMGA2 gene was constructed,and transfected into LS174T cells.The expression of HMGA2 mRNA and protein was detected by real-time PCR and Western blotting respectively.Cell proliferation was measured by MTT assay.The invasion ability of LS 174T cells was assessed by invasion assay with Transwell chamber.Results HMGA2 shRNA significantly inhibited the expression of HMGA2 in LS 174T cells at both mRNA and protein levels（P 0.01）.Compared with non interference group and negative control group,the proliferation rate of HMGA2shRNA-transfected LS 174T cells was decreased（P 0.05）.The abilities of invasion in transfected cells was inhibit as compared with the blank group（53.35 ± 8.45 vs.124.28 ± 65,P 0.05） and negative control group（53.35 ± 8.45 vs.118.52 ± 15,P 0.05）.Conclusion Inhibition of HMGA2 expression by shRNA significantly suppresses the proliferation and invasion of LS 174T cells in vitro.