摘要
为探讨卵巢癌细胞中TGF β的信号传导情况及TGF β/Smads信号通路各组分在卵巢癌发生中的作用 ,采用MTT和活细胞计数方法研究了TGF β1对卵巢癌细胞系HO 8910、HO 8910PM及SKOV3的生长抑制作用 ;并应用RT PCR、荧光免疫组化等方法研究了TGF β/Smads传导通路中各组分的表达和定位以及TGF β1刺激前后Smad7和P Smad2定位及表达的变化。结果显示 ,TGF β1对细胞系SKOV3没有生长抑制作用 ,而SKOV3细胞表达了TGF β/Smads信号通路中的所有已知成分。且 3种卵巢癌细胞在TGF β1刺激后Smad7mRNA瞬时表达增加 ,Smad7蛋白表达亦增加并由胞核转位到胞浆 ,P Smad2由胞浆转位到胞核。结果表明TGF β/Smads信号传导通路在卵巢癌细胞HO 8910、HO 8910PM和SKOV3中是完整的 ,SKOV3细胞逃逸TGF β介导的生长抑制作用可能是由于TGF
Resistance to the growth inhibitory effects of transforming growth factor-β(TGF-β) is a characteristic of many transformed cells. The purpose of this study was to determine the response of ovarian cancer cells to TGF-β1 and to investigate the roles of components of the TGF-β/Smads signaling pathway in carcinogenesis of ovarian cancer. Three ovarian cancer cell lines,HO-8910,HO-8910PM and SKOV3,were treated with TGF-β1 and assayed for growth response by MTT assay. Furthermore,expression and subcellular localization of the components of TGF-β/Smads signaling pathway in these cell lines in the absence or presence of TGF-β1 were determined by RT-PCR and immunofluorescence analysis. We found that proliferation of SKOV3 cell was not significantly inhibited by TGF-β1 while it expressed all components of the TGF-β/Smads signaling pathway. After exposure to TGF-β1,Smad7 protein in SKOV3 increased transiently and translocated to cytoplasm from nucleus while P-Smad2 translocated into nucleus from cytoplasm. Taken together,the results suggested that the TGF-β/Smads signaling pathway remained functional in human ovarian cancer cells,HO-8910,HO-8910PM and SKOV3,and the abnormalities of the downstream effectors of Smads proteins might contribute to the resistance of SKOV3 cell to TGF-β1.
基金
高等学校优秀青年教师教学科研奖励计划
国家自然科学基金 (编号 :3 0 3 70 783 )
黑龙江省科学技术计划攻关项目 (编号 :GB0 3C60 1-1)资助~~
