干扰Bmi-1表达对宫颈癌Hela细胞TGF-β/Smads通路基因表达的影响

Impact of Bmi RNAi on TGF-β/Smads Pathway in Hela cells

[目的]干扰Bmi-1对siRNA转染宫颈癌细胞系Hela中TGF-β、Hey1、Hes1、Bmp7、Smad3、Casp3和Casp6基因的表达影响,为寻找宫颈癌中Bmi-1的靶基因奠定基础。[方法]将Bmi-1 siRNA转染Hela细胞,实时荧光定量RT-PCR方法检测转染前后Hela细胞中TGF-β、Hey1、Hes1、Bmp7、Smad3、Casp3和Casp6基因的表达变化,对有明显变化的基因用WesternBlot在蛋白水平进一步验证。[结果]Hela细胞中Bmi-1基因RNA干扰后,TGF-β表达上调,Smad3、Casp3和Casp6基因表达下调。[结论]沉默Bmi-1的表达可使宫颈癌细胞系Hela中TGF-β、Smad3、Casp3和Casp6的表达量均有不同程度的改变,Bmi-1基因在宫颈癌发生发展中可能与TGF-β/Smads信号通路有关。

Purpose To evaluate the effect of Bmi RNAi on TGF-β,Hey1,Hes1,Bmp7,Smad3, Casp3 and Casp6 expressions in cervical cancer cell line Hela,in order to look for a basis for Bmi-1 targeted tumor gene in cervical cancer. Methods Hela cells were transfected with siRNA,then Real-time quantitative RT-PCR was used to detect the expression of TGF-β,Hey1,Hes1,Bmp7, Smad3,Casp3 and Casp6 in Hela cells. The genes with different expression after RNAi were validated at the protein level by Western blot.Results RNAi caused upregulation of TGF-β and downregulation of Smad3,Casp3 and Casp6 genes. Conclusions Bmi-1RNAi leads to change expression levels of TGF-β,Smad3,Casp3 and Casp6 in a definite level,It suggests Bmi-1 gene might play a role in carcinogensis for cervical cancer by acting on the TGF-β/Smads signaling pathway.