肝癌衍生生长因子对前列腺癌细胞增殖的影响及其机制  

Effects and mechanisms of hepatoma-derived growth factor on the proliferation of prostate cancer cells

作　　者：杨飞龙[1,2] 周尊林 任巨超[1] 闫磊[1] 刘海南[1] 张温花[1] 俞能旺[1] 李大伟 徐忠华[1] 

机构地区：[1]山东大学齐鲁医院泌尿外科,山东济南250012 [2]山东大学心血管重构与功能研究实验室,山东济南250012

出　　处：《山东大学学报（医学版）》2018年第1期62-69,共8页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81502213;81372335);山东省自然科学基金(ZR2015HM046);山东省重点研发计划(2016GSF201171)

摘　　要：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 blotting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(PAKT)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; ②Experimental 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, phosphatidylinositol 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 ceils, 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.

关 键 词：肝癌衍生生长因子 前列腺肿瘤 细胞增殖 P13K—AKT—mTOR信号通路 DU145 LNCAP 

分 类 号：R737.25[医药卫生—肿瘤]