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  

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

作　　者：肖蔚[1] 焦霞[1] 钱华[2] 崔永安[3] 林梅[3] 王薇[1] 周彤敏 窦荣荣 于鸿[1] 

机构地区：[1]泰州市人民医院病理科,江苏泰州225300 [2]泰州市人民医院妇产科,江苏泰州225300 [3]泰州市人民医院肿瘤科,江苏泰州225300

出　　处：《江苏大学学报（医学版）》2012年第5期382-386,共5页Journal of Jiangsu University：Medicine Edition

基　　金：江苏省自然科学基金资助项目(BK2010357);南通大学自然科学基金资助项目(10Z088)

摘　　要：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。Objective： To investigate the effect of connective tissue growth factor （CTGF） on the prolif- eration 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 quan- titive 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-tetrazolium- bromide （MT-F） method. Results： Positive clone C-16 with CTGF over-expression were successfully estab- lished. 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.

关 键 词：结缔组织生长因子 基质金属蛋白酶-2 基质金属蛋白酶-9 细胞增殖 人宫颈癌细胞 

分 类 号：R737.33[医药卫生—肿瘤] R456[医药卫生—临床医学]