RNA干涉技术稳定抑制PC-1基因表达对前列腺癌细胞C4-2的影响  

Impact of PC-1 Gene Knockdown on the Biological Action of Prostate Cancer Cell Line C4-2

作　　者：周立权[1] 张惠[1] 高雪松[2] 王健[1] 梁瑞霞[1] 洪宝发[2] 周建光[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]解放军总医院泌尿外科,北京100853

出　　处：《中华男科学杂志》2005年第4期256-260,共5页National Journal of Andrology

基　　金：国家"863"计划资助项目(2002AA223061);国家自然科学基金项目(30070296)

摘　　要：目的:研究用RNA干涉技术(RNAi)稳定抑制PC-1基因表达对前列腺癌细胞C4-2生物学行为的影响。方法:用DNA重组技术构建携带与短的发夹结构的干涉RNA相对应DNA序列的重组质粒pSC394-414,采用脂质体将pSC394-414重组质粒稳定转染C4-2细胞,PCR分析外源DNA序列的整合情况,RTPCR及Western印迹鉴定RNAi抑制PC1基因表达的细胞株,MTT试验分析该细胞的生长速度,软琼脂克隆形成实验检测其非锚着依赖性生长能力。结果:获得了用RNAi稳定抑制PC-1基因表达的细胞株C4-2-pSC394-414,该细胞株中的PC-1mRNA及PC1蛋白的表达均显著降低,细胞生长速度显著减慢,软琼脂克隆形成能力显著下降。结论:抑制PC-1基因表达,可降低C4-2细胞生长速度及非锚着依赖性生长能力。Objective: To study the effect of PC-1 gene knockdown on the biological action of prostate cancer cell line C4-2. Methods: Recombinant plasmids of expressing short hairpin RNA targeting PC-1 mRNA were constructed using DNA recombinant technology and transfected into C4-2 cells via liposome. The positive cell clones were selected by G418. The expression of PC-1 gene was analyzed by RT-PCR and Western blotting technology. MTT and soft agar cloning formation were applied to observe the changes of the growth rate and independent anchor ability of C4-2 cells. Results: PC-1 RNA interference severely affected the expression of PC-1 gene and reduced the growth and colony formation ability of C4-2 cells. Conclusion: RNA interference-mediated PC-1 gene knockdown can decrease the growth and cloning formation ability of C4-2 cells. Natl J Androl,2005,11(4):256-260

关 键 词：前列腺癌 PC—1基因 RNA干涉 基因表达 

分 类 号：R737.25[医药卫生—肿瘤] Q786[医药卫生—临床医学]