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目的 探讨人肝癌衍生生长因子(hHDGF)对前列腺癌细胞增殖的影响及相关机制.方法 根据hHDGF的干扰靶点构建短发卡RNA(shRNA)慢病毒载体,包装并纯化后稳定转染前列腺癌细胞株DU145和LNCaP.细胞分为2组:① 含无意义序列的慢病毒转染细胞作为对照组;② 含靶序列的慢病毒转染细胞作为实验组.采用实时荧光定量PCR(qRT-PCR)检测hHDGF mRNA表达,CCK-8实验和集落形成实验检测细胞增殖,Western blot-ting检测细胞内相关蛋白的表达水平.结果 成功构建慢病毒载体并稳定转染至前列腺癌细胞株,qRT-PCR和Western blotting结果示,hHDGF mRNA和蛋白表达均下降(P<0.05),CCK-8实验和集落形成实验均显示细胞增殖受到抑制(P<0.05),Western blotting结果示磷脂酰肌醇3激酶(PI3K)、磷酸化丝氨酸苏氨酸蛋白激酶(P-AKT)和磷酸化哺乳动物雷帕霉素靶蛋白(P-mTOR)的表达均下降(P均<0.05).结论 hHDGF表达下调对前列腺癌细胞增殖具有抑制作用,该作用可能与PI3K-AKT-mTOR信号通路改变相关.“,”Objective To study the effect and mechanism of human hepatoma-derived growth factor ( hHDGF) on the proliferation of prostate cancer cells. Methods Lentivirus vector was constructed according to the target sequence of hHDGF gene, which was then packaged into lentivirus particles and purified to transfect prostate cancer cells DU145 and LNCaP. The cells were divided into two groups separately:① Control group, which was treated by lentivirus with the insignificant sequence;② Experimental group, which was treated by the lentivirus with the target sequence. Relative mRNA expression of hHDGF was evaluated by quantitative real-time polymerase chain reaction ( qRT-PCR) . CCK-8 assay and colony formation assay were performed to measure cell proliferation ability. Relative expressions of hHDGF, phos-phatidylinositol 3-kinase ( PI3K) , phospho-serine/threonine protein kinase ( P-AKT) and phosphorylated mammalian target of rapamycin ( P-mTOR) proteins were detected by Western blotting. Results Lentivirus vector was successfully constructed, packaged and stably transfected into prostate cancer cells. qRT-PCR and Western blotting revealed that the mRNA and protein expression of hHDGF in experimental group were both downregulated ( P<0.05) . CCK-8 assay and colony formation assay indicated that cell proliferation of experimental group was inhibited ( P<0.05) . Western blotting revealed that relative protein expression of PI3K, P-AKT and P-mTOR were all downregulated ( P<0.05) . Conclusion Cell proliferation of proatate cancer is inhibited by the downregulation of hHDGF expression, which may be related with the inhibition of PI3K-AKT-mTOR signaling way.