TIMP-1基因体外稳定转染对人卵巢癌A_(2780)细胞周期与增殖的影响

Effect of TEMP-1 Gene Steadily Transfected on the Cell Cycle and Proliferation of A_(2780) Human Ovary Carcinoma Cells

目的:探讨正、反义基质金属蛋白酶组织抑制因子-1(TIMP-1)基因对人卵巢癌细胞周期及增殖的影响。方法:将重组质粒正、反义TIMP-1表达载体体外转染到卵巢癌A2780细胞中,用G418筛选稳定表达细胞株并扩增培养,采用MTT细胞计数法检测肿瘤细胞生长增殖率,流式细胞术检测肿瘤细胞周期的变化。结果:经15～20 dG418筛选得到稳定表达细胞株,重组质粒TIMP-1稳定转染正义(PTs)组A2780细胞的增殖可明显受到抑制,细胞周期明显改变,细胞从G1期到S期发生阻滞。而反义(PTas)组细胞与之相反。结论:提示体外转染TIMP-1基因可使A2780细胞周期阻滞于G1期,并可抑制肿瘤细胞增殖。可为临床基因治疗及相关基因药物研究提供实验依据。

Objective： To investigate the possible effects of tissue inhibitor of metalloproteinase-1 （ TIMP-1 ） gene on the cell cycle and proliferation of ovary carcinoma （A2780）. Method： A2780 cells were transfected with recombinant plasmids-PTS（ sense human TIMP- 1cDNA） and PTas（ antisense human TIMP-1 cDNA）. The stable express cells were selected by G418 and proliferated through cultiva- ting. The proliferation rate of tumor cells were examined by MTF cell counter and the changes of cell cycle were detected by FCM. Resuit： After 15 -20 daysG418 selection, the stable express cells were obtained. The growth of A2780 transfected with PTs was inhibited significantly and the cell cycle progression was arrested from GI to S phase. Those of PTas are contrary. Conclusion： These results suggest that TIMP-1 can arrest the cell cycle of A2780 at G1 phase and it can be significantly inhibited. The conclusion contributes to the clinical gene-therapy and the study on gene drugs.