TIMP-1对人卵巢癌A2780细胞生长的抑制作用

Growth Inhibition Effect of TIMP-1 on human gramulosa carcinoma A2780

关键词:  TIMP-1  卵巢癌  基因转染

Key words:  TIMP-1  gramulosa carcinoma  gene transfection

摘要:

［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T IMP-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TI MP-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。

Abstract:

［Objective］ To observe the effects on growth of A₂₇₈₀cells by TIMP-1 gene transfection. ［Methods］Transferred sense and antisense TIMP -1 into A₂₇₈₀gramulosa carcinoma cells by liposomal transfection reagent . Then we screened stable expression sublines by G418 solution and further confirmed by RT-PCR。Then the subtypes of A₂₇₈₀ were marked as 2780-PTs,2780-Pt as and A-2780， respectively. We also evaluated the proliferation activities in these three subtypes by direct cells count，MTT assay, plate clone formation tes t，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 G0/G1 decrease (44.82±0.31)%.［Conclusion］TIMP-1 transfection can inhibit the growth of A₂₇₈₀ gramulosa carcinoma cell.ThusTIMP-1 can be a novel target for gramulosa carcinoma gene therapy.