TIMP-1对人卵巢癌A_(2780)细胞生长的抑制作用

Growth Inhibition Effect of TIMP-1 on human gramulosa carcinoma A_(2780)

[目的]观察基质金属蛋白酶组织抑制因子-1(TIMP-1)基因表达水平对人卵巢癌细胞生长的影响。[方法]采用质脂体法将正、反义TIMP-1表达载体体外转染到卵巢癌A2780细胞中,用G418筛选稳定表达外源基因的克隆并用RT-PCR鉴定后扩大培养。以未转染组细胞作为对照,分别用直接计数法、四氮唑蓝(MTT)比色法、平皿克隆形成试验测定3组细胞的增殖活性,利用流式细胞术检测肿瘤细胞周期与凋亡的变化。[结果]与对照组相比,转染TIMP-1对卵巢癌细胞A2780呈现增殖抑制效应:生长曲线示增殖速度减慢(P<0.01);在平皿上的集落形成能力下降(38±3.05)%;MTT法结果显示增殖抑制率增加(P<0.01);细胞周期显示G0/G1期比例增加(58.41±0.94)%。转染反义TIMP-1对卵巢癌细胞A2780呈现促增殖效应:生长曲线示增殖速度加快(P<0.01);在平皿上的集落形成能力增强(59±2.08)%;MTT显示增殖抑制率减小(P<0.01);细胞周期显示G0/G1期比例减小(44.82±0.31)%。[结论]体外转染TIMP-1基因提高TIMP-1的表达可抑制卵巢癌细胞的增殖,TIMP-1可能成为卵巢癌基因治疗的一个新靶点。

[ Objective] To observe the effects on growth of A2780 cells by TIMP - 1 gene transfection. [ Methods] Transferred sense and antisense TIMP- 1 into A2780 gramulosa carcinoma cells by liposomal transfection reagent. Then we screened stable expression sublines by G418 solution and further confirmed by RT - PCRo Then the subtypes of A2780 were marked as 2780 - PTs,2780 - Ptas and A - 2780, respectively. We also evaluated the proliferation activities in these three subtypes by direct cells count, MTT assay, plate clone formation test, respectively;The change of cell cycle were detected by FCM. [ Results] Compared with the normal A2780, 2780- PTs showed sush characteristics as followed： Partial inhibition of proliferation activities reflected by growth curves（ P 〈 0.01 ） ; Decreasing clone formation on plate（38 ± 3.05）%;the proliferation inhibition rate increase revealed by MTT method result（ P 〈 0.01 ）; cell cycle reflected G0/G1 increase（58.41 ± 0.94）%. On the contrary, the 2780- PTas subline showed promoted proliferation, which was indicated as followed： Promoted proliferation activities under cells culture（ P 〈 0.01 ） ; Increasing clone formation on plate（59 ± 2.08） % ;decreaseing proliferation inhibition rate revealed by MTT method result （P 〈 0.01 ）;cell cycle reflected Go/Gl decrease （44.82 ± 0.31）%. [Conclusion] TIM P- 1 transfection can inhibit the growth of A2780 gramulosa carcinoma cell. ThusTIMP- 1 can be a novel target for gramulosa carcinoma gene therapy.