HPV16E6和E7对乳腺癌细胞侵袭能力的影响及其机制

目的探讨HPV16E6、E7和E6/E7同时稳定高表达对乳腺癌细胞MCF-7侵袭能力的影响及其机制。方法利用脂质体介导将HPV16E6、E7和全长E6/E7转染MCF-7细胞,利用Western blot检测转染后E6和E7的表达。采用Transwell实验检测MCF-7细胞侵袭能力的变化,RT-PCR法检测基质金属蛋白酶-2(MMP-2)mRNA的表达情况。结果在MCF-7-E6细胞克隆中,克隆1表达水平最高;MCF-7-E7细胞克隆中,克隆4表达水平最高;MCF-7-E6/E7细胞克隆中,克隆4表达水平最高。与MCF-7亲本和MCF-7-vect细胞比较,MCF-7-E6和MCF-7-E6/E7细胞穿过小室底膜的细胞数均显著增加(P<0.01)。在MCF-7-E6、MCF-7-E7和MCF-7-E6/E7细胞中MMP-2mRNA表达均显著增加。结论 HPV16E6和E7促进MCF-7细胞体外侵袭能力的机制之一可能是通过诱导MMP-2的表达。

Effects of HPV 16 E6 and E7 on invasion of carcinoma cell and its mechanism

Objective To investigate the effects of HPV 16 E6,E7 and simultaneous expression of E6 and E7 on the invasion of breast carcinoma cells MCF-7 and its mechanism.Methods The recombinant plasmids were transfected into human breast carcinoma cell MCF-7 mediated by lipofectamine.The Western blotting was used to detect the effects of stable expression of HPV16 E6,E7 and simultaneous expression of E6 and E7.The invasive ability was investigated by Transwell assay.The mRNA expression of MMP-2 was detected by RT-PCR.Results The results from Western blotting indicated that protein expression of clone 1 was highest in the four MCF-7-E6 positive clones,and protein expression of clone 4 was highest in the four MCF-7-E7 positive clones,and expression of clone 4 was highest in the four MCF-7-E6/E7 positive clones.The results from Transwell assay disclosed that compared with MCF-7 parental cells,MCF-7-vect cells and MCF-7-E7 cells,the numbers of invasion and metastasis cells were significantly increased in MCF-7-E6 cells and MCF-7-E6/E7 cells(P<0.01).The RT-PCR results indicated that expression of MMP-2 mRNA in MCF-7-E6 cells,MCF-7-E7 cells and MCF-7-E6/E7 cells were significantly increased than that in parental cells and MCF-7-vect cells.Conclusion HPV16 E6 and E7 can promote the expression of MMP-2,which may be one of its mechanisms of promoting invasion ability of MCF-7 cells.