摘要
目的探讨RNA干扰技术(RNAi)封闭人前列腺癌细胞系PC-4中Smad4的表达以阻断转化生长因子(TGF)-β/Smad信号转导进而诱导细胞凋亡的机制。方法以RNAi特异性封闭PC-4中Smad4的表达,逆转录-聚合酶链反应(RT-PCR)及Western blot分别检测Smad4的表达,RT-PCR检测TGF-β/Smad信号通路下游分子Smad2/3的表达,并绘制细胞生长曲线。结果自RNAi后12h起,PC-4细胞中Smad4 mRNA表达量为封闭前的20%～68%,蛋白表达量为封闭前的8.0%～17.2%,Smad2/3 mRNA表达量为封闭前的0%～52%,且细胞凋亡加速。结论抑制Smad4表达可以在一定程度上阻断PC-4细胞TGF-β/Smad信号的转导,进而促进细胞凋亡。
To investigate the mechanism of blocking the transduction of transforming growth factor （TGF） --β/Smad signal pathway in human prostate cancer cell lines in PC-4 by use of RNA interference （RNAi） and apoptosis induction of PC cells. Methods The expression of Smad4 in PC-4 cell lines was specifically blocked by RNAi. RT-PCR and Western blot were used to detect the expression of Smad4. The downstream molecule, Smad2/3, was detected by RT-PCR and the growth curves of PC cells were drawn. Results Twelve h after blocking by RNAi, the expression of Smad4 mRNA and protein and Smad2/3 mRNA in PC cells was respectively 20% -68% , 8% -17.2% and 0-52% of that before blocking. Furthermore, RNAi of Smad4 could even accelerate the apoptosis of PC ceils. Conclusion Suppression of Smad4 by RNAi can efficiently block the TGF-β/Smad signal of PC-4 cells, and subsequently enhance the apoptosis of PC ceils.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第10期1176-1178,共3页
Chinese Journal of Experimental Surgery
关键词
前列腺癌
转化生长因子-Β
转染
RNA干扰
Prostate carcinoma
Transforming growth factor-13
Transfect
RNA interference
