摘要
目的:探讨RNA干扰技术沉默STAT3基因表达对人前列腺癌细胞PC3及LNCaP的生长抑制作用.方法:针对STAT3 mRNA序列设计合成3对编码小干扰RNA(siRNA)的DNA模板,构建pSilencer 1.0-U6-siRNA-STAT3重组质粒,转染PC3及LNCaP细胞;采用Western印迹、Northern印迹等技术观察重组质粒对STAT3基因表达的影响;用MTT法观察重组质粒对PC3及LNCaP细胞体外生长抑制作用;用流式细胞术(FCM)及吖啶橙染色检测重组质粒诱导细胞凋亡.结果:成功构建pSilencer1.0-U6-siRNA-STAT3重组质粒,并成功转染PC3及LNCaP细胞;Western印迹,Northern印迹结果证实重组质粒在mRNA及蛋白水平分别显著抑制STAT3基因表达,抑制率为60%～75%;MTT及FCM结果证明上述重组质粒可显著抑制PC3及LNCaP细胞的体外生长并诱导PC3细胞凋亡.结论:pSilencer1.0-U6-siRNA-STAT3可抑制STAT3在人前列腺癌细胞中的表达,并抑制肿瘤细胞的生长,促进其凋亡.
Objective: To study the effects of pSilencer 1.0-U6-siRNA-STAT3 on the growth of PC3 and LNCaP cells. Methods: Three pairs of DNA template coding siRNA were synthesized against STAT3 to reconstruct pSilencer 1.0-U6-STAT3-siRNA, which was transfected into PC3 and LNCaP cells. The STAT3 expression in PC3 cells and LNCaP were transfected with pSilencer 1.0-U6-siRNA-STAT3, and it was detected by Western blot and Northern blot. MTT and FCM were used to observe the growth-inhibiting ratio and apoptosis in PC3 cells. Results: Western blot and Northern blot analyses demonstrated that pSilencer 1.0-U6-siRNA-STAT3 could significantly inhibit the expression of STAT3 in PC3 and LNCaP cells; MTT and FCM results showed that it could suppress the growth of PC3 cells and LNCaP and induce apoptosis of PC3 cells in vitro. Conclusion: PSilencer 1.0-U6-siRNA-STAT3 could significantly inhibit STAT3 expression, suppress the growth of PC3 and LNCaP cells and induce the apoptosis of PC3 cells. Natl J
出处
《中华男科学杂志》
CAS
CSCD
2005年第1期29-33,37,共6页
National Journal of Andrology
基金
中日政府间专项技术合作项目第 59项(1999)
