Objective To investigate the effect and mechanism of Androgen receptor(AR)small hairpin RNA(shRNA) plasmid on prostate cancer xenografts. Methods Recombinant plasmid of AR shRNA was constructed and prepared. The BALB/C nude mice were used to construct DU-145 prostate cancer cell xenografts model and divided into model control,positive control and experimental groups(n=10). The positive control group, the experimental group and the model control group were inoculated with Fluorouracil (20 mg/kg), AR shRNA plasmid (2 mg/kg) and normal saline for 10 days respectively to determine the tumor volume. The growth of nude mice was sacri-ficed at 4 weeks after the start of administration. AR mRNA expression in xenografts was detected by RT-PCR. The proteins in PI3K/AKT signal pathway was detected by Western blot. Results There was no significant difference in the volume of the transplanted carcinoma be-tween the experimental and the model control groups after administrate 3 days(P>0.05). The tumor volume of the experimental group at 1 week,2 weeks,3 weeks and 4 weeks after administration was significantly reduced than that in the model control group(P<0.05). The ex-pression of AR mRNA in the transplanted carcinoma of nude mice in experimental group was significantly lower than that in model control group (P<0.05). The expression of p-AKT,p-GSK3β and p-mTOR proteins was significantly lower than that of the model control group(P<0.05). The expression of AKT,GSK3β and mTOR protein was significantly higher than that of the model control group(P<0.05).Conclu-sions AR-shRNA could inhibit the growth of prostate cancer xenografts by inhibiting the expression of androgen receptor and the phosphoryl-ation of PI3K/AKT signaling pathway in prostate cancer.%目的 探讨雄激素受体(AR)小片段发夹 RNA(shRNA)质粒对前列腺癌移植瘤的作用及其机制.方法 构建AR shRNA并制备重组质粒.使用BALB/C裸鼠构建前列腺癌细胞DU-145移植瘤模型并分为模型对照组、阳性对照组和实验组(n=10).阳性对照组、实验组和模型对照组裸鼠分别尾静脉注射氟尿嘧啶(20 mg/kg)、AR shRNA质粒(2 mg/kg)及等体积生理盐水共10 d,测定各组裸鼠的肿瘤体积生长情况,开始给药后第4周处死裸鼠剥离肿瘤,采用RT-PCT测定移植瘤中AR mRNA表达水平,采用Western印迹测定磷脂酰肌醇-3-羟激酶(PI3K)/丝氨酸-苏氨酸蛋白激酶(AKT)信号通路相关蛋白表达水平.结果 给药3d时实验组和模型对照组裸鼠肿瘤体积差异无统计学意义(P>0.05),实验组裸鼠在给药后1w、2w、3w和4w时肿瘤体积较模型对照组显著降低(P<0.05).实验组裸鼠移植瘤AR mRNA表达水平较模型对照组显著降低(P<0.05).p-AKT、p-GSK3β和p-雷帕霉素靶蛋白(mTOR)蛋白表达水平较模型对照组显著降低(P<0.05),AKT、GSK3β和mTOR蛋白表达较模型对照组显著升高(P<0.05).结论 AR shRNA可抑制前列腺癌组织AR表达及PI3K/AKT信号通路相关蛋白磷酸化而发挥对前列腺癌移植瘤生长的抑制作用.
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