锤头型核酶特异性阻断雄激素受体表达对前列腺癌细胞生长的影响

目的：探讨核酶阻断雄激素受体（AR）基因表达治疗前列腺癌的可能性。方法：在脂质体介导下，锤头型抗雄激素受体核酶表达载体转染前列腺癌细胞，采用逆转录－聚合酶链反应（RT－PCR）检测ARmRNA，四甲基偶氮唑盐（MTT）法检测细胞增殖活性，流式细胞仪（FCM）检测细胞周期时相，原位末端转移酶标记法检测细胞凋亡。结果：核酶表达载体转染1－5d后，前列腺癌细胞AR mRNA水平降低32．6％－40．7％（P＜0．05），细胞生长抑制率18．28％－35．34％（P＜0．05），细胞周期阻滞于G2／M期，诱导细胞凋亡，细胞凋亡比率增加20．70％（P＜0．01）。结论：应用核酶特异性切割ARmRNA，能有效阻断AR基因的表达，诱导前列腺癌细胞凋亡，有可能成为前列腺癌基因治疗的有效方法之一。

Effects of blocking androgen receptor expression by specific hammerhead ribozyme in vitro on prostate cancer cell growth

Objective To explore the possibility of hammerhead ribozyme gene therapy for prostate cancer.Methods The vector pcDNA hAR RZ expressing specific hammerhead ribozyme against androgen receptor was transformed into prostate cancer cell line (LNCaP) by lipofectamine.Androgen receptor mRNA was detected by RT-PCR and cell growth and apoptosis were studied by MTT,FCM and TUNEL.Results AR mRNA was decreased by 32.6 % to 40.7 %(P<0.05),with cell growth being inhibited by 18.28 % to 35.34 % (P<0.05) after pcDNA-hAR-RZ transfection for 24 to 72 h,LNCaP cells were blocked at G 2/M stage and apoptosis was induced by ribozyme with the apoptosis rate being increased by 20.7 % as compared with that in the control group.Conclusion The specific hammerhead ribozyme against androgen receptor was capable of inhibiting the expression of androgen receptor and inducing apoptosis of LNCaP cells.It might be used an effective way for prostate cancer gene therapy.