TGF-β1和Smad4基因转染对子宫内膜癌细胞HHUA体外增殖和凋亡的研究  

Study of TGF-β1 and Smad4 gene transfection on the proliferation and apoptosis of endometrial carcinoma cell line HHUA in vitro.

作　　者：纪国欣[1] 王欣[1] 赵兴波[1] 李明江[1] 李继俊[1] 

机构地区：[1]山东大学附属山东省立医院妇产科

出　　处：《现代妇产科进展》2007年第11期822-826,882,共6页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目(No:30640076)

摘　　要：目的:探讨TGF-β1和Smad4基因转染对子宫内膜癌细胞HHUA体外增殖、细胞周期、凋亡及Smad7、PAI-1的影响。方法:电穿孔法将Smad4基因转染入HHUA细胞。观察未转染和转染Smad4基因的HHUA细胞在不同浓度TGF-β1(0,2.5,5,10ng/ml)培养液中其Bcl-2、Bax、Smad7、PAI-1的表达、肿瘤细胞增殖活性和凋亡活性变化及Smad7、PAI-1和TGF-β1 mRNA水平。结果:TGF-β1以时间依赖方式明显抑制HHUA生长(P<0.05),各浓度组间无显著差异;转染Smad4后TGF-β1仍明显抑制HHUA生长(P<0.01)。转染Smad4前后,TGF-β1均阻滞细胞停滞于G1期;转染后作用增强,差异有统计学意义(P<0.01)。TGF-β1明显降调Bcl-2,升调Bax、Smad7和PAI-1表达,促进细胞凋亡;转染Smad4后作用增强(P<0.05)。TGF-β1、Smad7和PAI-1 mRNA水平在TGF-β1作用和(或)Smad4转染前后都明显升高。结论:TGF-β1以时间依赖方式抑制HHUA体外增殖,阻滞细胞周期,促进细胞凋亡;转染Smad4明显增强了TGF-β1的抗肿瘤作用。To investigate the effect of TGF-β1 and Smad4 gene on the proliferation, cell cycle, apoptosis and the change of Smad7 and PAI-1 of the endometrial carcinoma cell line HHUA in vitro. Methods：Electroporation was used to transfect the Smad4 gene into HHUA in vitro. The transfected and non-transfected HHUA cells were cultured in the media containing TGF-β1 of different concentration （0,2.5,5,10ng/ml）. The expression of Bcl-2, Bax,Smad7 and PAI-1 was measured by immunohistochemestry, the proliferation and phase of cell cycle was detected by MTY and FCM, the semi-quantitative RT-PCR was adopted to measure the mRNA of Smad7, PAI-1 and TGF-β1. Results：TGF-β1 inhibited the proliferation of HHUA ,and it had no significant difference with different concentration of TGF-β1. The effect of TGF-151 was significantly enhanced after transfecting Smad4 gene. TGF-β1 blocked the HHUA cell at G1 phase, and its effect was enlarged after transfection of Smad4 gene. TGF-β1 could down-regulate Bcl-2 expression,up-regulate Bax, Smad7 and PAI-1 expression, and could promote the apoptosis of HHUA in time dependent fashion. The mRNA level of Smad7 ,PAI-1 and TGF-β1 was elevated after TGF-β1＇s effect and/or transfection of Smad4 gene, comparing with non-transfection of Smad4 gene. Conclusion： TGF-β1 can inhibit the proliferation, block cell cycle,and promote the apoptosis of HHUA in time-dependent fashion. The effect of TGF-β1 is significantly raised after transfecting Smad4 gene in vitro.

关 键 词：子宫内膜肿瘤 转化生长因子Β1 基因 SMAD4 细胞凋亡 细胞系 

分 类 号：R737.33[医药卫生—肿瘤]