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人核小体结合蛋白1小分子干扰RNA重组慢病毒抑制非激素依赖性前列腺癌体内生长的实验研究

蒋宁,周利群,姚鲲,黄晨

蒋宁、周利群、姚鲲、黄晨,北京大学泌尿外科研究所 北京大学第一医院泌尿外科,100034

基金项目:国家自然科学基金(30271295,30672099);北京市自然科学基金(7092101)

摘 要:目的 探讨重组慢病毒介导的人核小体结合蛋白1(NSBP1)基因沉默对非激素依赖性前列腺癌细胞系DU145移植瘤生长的抑制作用及机制。方法 慢病毒lentivirus-NSBP1转染非激素依赖性前列腺癌细胞系DU145。用转染细胞成瘤,观察抑瘤效果。运用实时RT-PCR和Western blot方法检测移植瘤中NSBP1、cyclinB1与Bcl-2的mRNA和蛋白的表达。结果 体内实验证实NSBP1表达水平的降低,对肿瘤细胞在裸鼠体内成瘤率没有影响,但对肿瘤的生长有明显抑制作用。同时随着NSBP1表达水平的降低,cyclinB1和Bcl-2的mRNA及蛋白的表达也降低。结论 NSBP1-RNAi-lentivirus 重组慢病毒能有效抑制DU145细胞移植瘤的生长,NSBP1可能通过调控cyclinB1、Bcl-2基因的变化影响肿瘤细胞的生长。

关键词:前列腺肿瘤; RNA,小分子干扰; HMGN1蛋白质; 慢病毒属; 印迹法,蛋白

The experimental investigation of androgen-independent prostate cancer cell line inhibited growth in vivo by recombinant lentivirus of small interfering RNA(siRNA)targeting NSBP1

JIANG Ning,ZHOU Li-qun,YAO Kun,HUANG Chen.

Department of Urology,Peking University First Hospital,Institute of Urology,Peking University,Beijing 100034,China

Abstract:Objective To study the effect and mechanism of constructed a lentiviral vector harboring RNAi sequence targeting the human high mobility group NSBP1 gene on the growth of tumor transplanted from human androgen-independent prostate cancer cell DU145 into nude mice. Methods The transfected lentivirus-NSBP1 DU145 cells were transplanted into nude mice,then tumor inhibitive effect was observed. Realtime RT-PCR and Western blot were used to test the expression of NSBP1,cyclinB1 and Bcl-2 in the xenografts. Results In an in vivo study,the downregulation of NSBP1 reduced cancer growth rate,whereas tumorgenicity was not influenced. After knockdown NSBP1,expressions of cyclin B1 and Bcl-2 mRNAs and proteins decreased. Conclusions The growth of xenografts could be inhibited by recombinant NSBP1-RNAi-lentivirus,NSBP1 may effect on the growth of the prostate cancer cell line DU145 resulting from regulating cyclin B1 and Bcl-2 expression.

Key words:Prostatic neoplasms; RNA,small interfering; HMGN1 protein; Lentivirus; Blotting,western

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文献标引:蒋宁,周利群,姚鲲,黄晨.人核小体结合蛋白1小分子干扰RNA重组慢病毒抑制非激素依赖性前列腺癌体内生长的实验研究[J/CD].中华临床医师杂志:电子版,2010,4(2):154-157.

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