RNA沉默ERp29基因促进人前列腺癌细胞体外侵袭及其机制研究  

Silencing ERp29 promotes the invasiveness of human PCa cells in vitro:Molecular mechanisms

作　　者：朱斌[1] 黄智超[1] 邓黄浩 董志韬[1] 赵晓昆[1] 杨罗艳[1] 蒋宏毅[1] ZHU Bin;HUANG Zhi-chao;DENG Huang-hao;DONG Zhi-tao;ZHAO Xiao-kun;YANG Luo-yan;JIANG Hong-yi(Department of Urology,The Second Xiangya Hospital,Central South University,Changsha,Hunan 410011,China)

机构地区：[1]中南大学湘雅二医院泌尿外科,湖南长沙410011

出　　处：《中华男科学杂志》2020年第7期579-587,共9页National Journal of Andrology

摘　　要：目的:对内质网分子伴侣29(ERp29)与前列腺癌患者的临床病理特征相关性进行综合分析以及RNA干扰ERp29基因对LNCaP前列腺癌细胞株侵袭的影响及其机制。方法:通过免疫组化法检测良性前列腺增生组织和前列腺癌组织中ERp29的表达,通过Western印迹方法检测6例前列腺癌组织以及临近正常组织中ERp29蛋白的差异性;通过脂质体转染法将特异性沉默ERp29基因的siRNA转染人LNCaP细胞;分别用实时荧光定量PCR(qRT-PCR)和Western印迹方法检测ERp29 siRNA对LNCaP细胞ERp29基因和蛋白表达的抑制作用;运用MTT法检测LNCaP细胞的增殖活力;利用Transwell法检测细胞体外迁移及侵袭能力;通过qRT-PCR法检测上皮-间质转化相关标志物变化。结果:前列腺癌中ERp29的高表达阳性率低于非肿瘤前列腺组织(73.9%vs 91.9%,P<0.05);前列腺癌组织中的ERp29低表达阳性率与M分期显著相关(P<0.05)。LNCaP细胞转染ERp29 siRNA后,其细胞增殖能力显著增强,迁移和侵袭能力也显著提高,差异有统计学意义(P<0.05);E-cadherin表达明显下降,Vimentin表达明显上升,与空白对照组相比差异有统计学意义(P<0.05)。结论:ERp29可能是一种新的肿瘤转移抑制基因,沉默ERp29基因能促进人前列腺癌细胞体外侵袭能力,其机制与下调E-cadherin,促进上皮-间质转化有关。Objective:To analyz the correlation of the expression of ERp29 with the clinicopathological characteristics of PCa and investigate the effect of small interfering RNA(siRNA)silencing the ERp29 gene on the biological behavior of PCa LNCaP cells.Methods:The expression of the ERp29 gene in the BPH and PCa tissues was detected by immunohistochemistry and that of the ERp29 protein in the PCa and adjacent normal tissues of 6 PCa patients determined by Western blot.Human LNCaP cells were transfected with siRNA using LipofectamineTM 2000,and the expressions of ERp29 mRNA and protein in the LNCaP cells detected by quantitative real-time PCR(qRT-PCR)and Western blot,respectively.The proliferation of the LNCaP cells was measured by MTT assay,their in vitro migration and invasiveness evaluated by the Transwell method,and the expressions of E-cadherin and Vimentin determined by qRT-PCR.Results:The expression of ERp29 was significantly lower in the PCa than in the adjacent normal tissue(73.9%vs 91.9%,P<0.05),with a significant correlation between the down-regulated ERp29 expression and metastasis(M)staging(P<0.05).After transfection with siRNA,the LNCaP cells showed dramatically increased proliferation,migration and invasiveness(P<0.05),and the expression of E-cadherin was markedly down-regulated while that of Vimentin up-regulated as compared with those in the normal control group(P<0.05).Conclusion:The ERp29 gene may be a novel repressor of tumor metastasis.Silencing ERp29 can promote the invasiveness of human PCa cells in vitro by down-regulating the expression of E-cadherin and increasing epithelial-mesenchymal transition.

关 键 词：前列腺癌 肿瘤转移 小分子干扰 上皮-间质转化 体外 

分 类 号：R737.25[医药卫生—肿瘤]