CTGF基因转染对宫颈癌细胞MMP-2、MMP-9表达及细胞增殖的影响

目的:研究结缔组织生长因子(connective tissue growth factor,CTGF)基因对人宫颈癌Hela细胞基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达及细胞增殖的影响,探讨CTGF在宫颈癌侵袭和转移中的作用机制。方法:经脂质体介导将含有CTGF重组表达质粒转染人宫颈癌Hela细胞株,用G418筛选阳性细胞克隆及实时荧光定量PCR、蛋白质印迹鉴定;采用实时荧光定量PCR和蛋白质印迹法检测阳性克隆细胞MMP-2及MMP-9的表达;噻唑盐(MTT)比色法检测阳性细胞克隆的增殖活性。结果:成功建立稳定高表达CTGF的阳性Hela细胞克隆,证实其MMP-2、MMP-9表达及细胞增殖活性均显著增高。结论:CTGF转染可显著增加宫颈癌细胞MMP-2及MMP-9的表达并促进其细胞增殖,CTGF可能是宫颈癌基因治疗的潜在靶点。

Effect of gene CTGF transfection on the expression of MMP-2 and MMP-9 and proliferation in human cervical cancer cells

Objective: To investigate the effect of connective tissue growth factor(CTGF) on the proliferation in human cervical cancer cell line Hela,with a focus on the expression of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9(MMP-9),and explore the underlying mechanism for the role of CTGF in the development of cervical cancer.Methods: pcDNA3.0-CTGF and pcDNA3.0 were transfected into Hela cells through lipofectamine and positive clones which were screened by G418.Fluorescence quantitive polymerase chain reaction(FQ-PCR) and Western blot were employed to identify mRNA and protein expression of CTGF in Hela cells,respectively.The expression of MMP-2 and MMP-9 in positive clones was detected by FQ-PCR and Western blot.Cell viability was assessed by dimethylthiazoldiphenyl-tetrazoliumbromide(MTT) method.Results: Positive clone C-16 with CTGF over-expression were successfully established.Compared with non-transfected control group,the expression of MMP-2 and MMP-9 in C-16 were increased significantly,and the proliferation level of C-16 was increased significantly.Conclusion: CTGF transfection could effectively enhance the expression of MMP-2 and MMP-9 and the proliferation in Hela cells which suggested a potential role for CTGF in gene therapy of cervical cancer.