利用RNA干涉技术研究雄激素受体在人前列腺癌细胞增殖中的作用  

Role of androgen receptor in prostate cancer cell proliferation by use of RNAi

作　　者：颜艳灵[1] 钟雪云[2] 钟振宇 秦艳芳[2] 洪岸[1] 陈弘武 

机构地区：[1]暨南大学生物工程研究所,广东广州510632 [2]暨南大学医学院病理学教研室,广东广州510632 [3]加洲大学戴维斯分校癌症中心,美国加洲95817

出　　处：《中国病理生理杂志》2004年第4期580-583,共4页Chinese Journal of Pathophysiology

基　　金：重大基础研究前期研究专项 (973前期预研;No.2 0 0 2ccc0 0 4 0 0 ) ;广东省科委科技攻关项目 (2 0 0 3B30 10 1)

摘　　要：目的 :利用RNA干涉技术抑制雄激素受体 (AR)的表达 ,研究AR在激素依赖性和激素非依赖性前列腺癌细胞增殖中的作用。方法 :设计、合成针对AR的 4种不同的小干涉RNA(siRNA) ,连接到带有人H1启动子的腺病毒载体质粒pShuttle -H1-Ri中 ,构建成能产生AR -siRNA的质粒pShuttle -H1-Ri-AR ,与能表达AR的质粒pcDNA -AR共转染HEK2 93细胞 ,Westernblot检测不同的siRNA对AR表达的抑制效率 ,选取抑制效率高的siRNA制备重组腺病毒 ,感染LNCap、C4 - 2B和CWR2 2Rv13种对雄激素有不同反应性的人前列腺癌细胞 ,采用Westernblot检测感染后细胞中AR的表达程度 ,并用MTT比色法测定细胞的增殖活性。结果 :4种siRNA都能抑制共转染AR的表达 ,用抑制效率高的siRNA制备重组腺病毒感染 3种靶细胞后 ,均能特异性地抑制 3种细胞中AR的表达 ,细胞的增殖活性也随之明显下降。结论 :AR -siRNA通过抑制激素依赖性和激素非依赖性前列腺癌细胞中AR的表达 ,有效地抑制细胞的增殖 ,AR对维持激素依赖性和激素非依赖性前列腺癌细胞的增殖活性具有十分重要的作用 ,是治疗前列腺癌的重要靶分子。AIM: To study the role of androgen receptor (AR) in hormone-dependent and hormone-independent prostate cancer cell proliferation by knocking down AR expression with adenovirus-delivered siRNA. METHODS: Four well-designed siRNAs were synthesized and inserted into the adenovirus plasmid pShuttle-H1-Ri. The recombinant pShuttle-H1-Ri-AR plasmid was then co-transfected with pcDNA-AR to HEK293 cell line and Western blot was used to detect the inhibitory efficiency of different siRNAs on AR expression. Recombinant adenovirus containing more efficient siRNAs were prepared and used to infect three different humane prostate cancer cell lines including LNCapC4-2B and CWR22Rv1. The efficiency of knocking down AR expression was detected by Western blot. The effect of AR-knocking down on cell proliferation was detected by MTT colorimetric assay. RESULTS: All of the four designed siRNAs could knock down AR expression in transient co-transfection. Infecting with recombinant adenovirus containing more efficient siRNAs in hormone-dependent and hormone-independent prostate cancer cell lines specifically knocked down AR expression with high efficiency. Knocking down AR expression significantly decreased the proliferation rate in all these prostate cancer cells. CONCLUSION: The suppressed expression of AR in prostate cell lines mediated by siRNA could efficiently inhibit the cell proliferation, and these results show that AR plays an important role in the proliferation of hormone-dependent and hormone-independent prostate cancer cells. AR is an important therapeutic target for the treatment of prostate cancer.

关 键 词：受体 雄激素 重组腺病毒 SIRNA 前列腺肿瘤 

分 类 号：Q754[生物学—分子生物学] R737.25[医药卫生—肿瘤]