慢病毒介导跨膜丝氨酸蛋白酶2∶ERG融合基因短发卡RNA靶向抑制前列腺癌细胞生长  

Lentivirus-mediated transmembrane protease serines 2∶ERG silence inhibits proliferation of human prostate cancer line VCaP in vitro

作　　者：梁辉[1] 张圣平[1] 张建文[1] 王宏亮[1] 孙睿[1] 植凡[1] 

机构地区：[1]南方医科大学附属深圳市龙华新区人民医院泌尿外科,深圳518019

出　　处：《中华实验外科杂志》2015年第6期1300-1302,共3页Chinese Journal of Experimental Surgery

基　　金：广东省医学科研基金资助项目（A2012603）

摘　　要：目的 观察靶向抑制跨膜丝氨酸蛋白酶2（TMPRSS2）：ERG融合基因后对前列腺癌细胞生长的影响.方法 设计特异性作用于TMPRSS2∶ ERG融合基因的短发卡RNA （shRNA）,并进行慢病毒包装后感染前列腺癌细胞,进一步通过噻唑蓝（MTT）实验,以及流式细胞术研究ERG基因沉默后前列腺癌细胞增殖的变化.结果 成功构建并筛选到高效特异性的shRNA1序列,在mRNA水平的靶向抑制效率为79％,蛋白水平的靶向抑制率为93％;靶向抑制TMPRSS2∶ ERG融合基因表达96 h后,前列腺细胞生长速度降低了33.34％;细胞周期分析发现ERG基因沉默后的VCaP细胞S期和G2/M期细胞比例分别由10.0％降至6.6％,29.1％下降到17.25％,而G0/G1期细胞比例由51.7％升高到71.2％.结论 靶向抑制TMPRSS2∶ ERG融合基因的表达可以显著抑制前列腺癌细胞的生长.Objective To study the effects of transmembrane protease serines 2 （TMPRSS2） ∶ ERG fusion gene silencing on the growth of prostate cancer VCaP cells.Methods TMPRSS2∶ERG fusion gene shRNAs were designed and constructed to lentiviral vector pLKO.1,virus packing by 293FT cells and infected into the VCaP cells.The TMPRSS2∶ ERG fusion gene silence efficiency was tested by real-time quantitative polymerase chain reaction（Real-time PCR） and Western blotting respectively.Methyl thiazol tetrazolium （MTT） and flow cytometry were used to examine the proliferation capacity and cell cycle of VCaP cells.Results Three pairs shRNAs specific to the ERG gene were successfully designed,and the shRNA-ERG lentiviral clones were constructed.The shRNA1 showed the highest inhibition efficiency.The ERG mRNA level was reduced by 79％,and ERG protein level by 93％ after shRNA1 lentivirus infection,respectively.The MTT results revealed that the proliferation capacity of VCaP cells was reduced by 33.34％ after TMPRSS2∶ ERG fusion gene was silenced.Cell cycle analysis demonstrated that the ratio of S phase cells was reduced to 6.6％ from 10.0％,that of G2/M phase cells was reduced to 17.25％ from 29.1％,and that of G0/G1 phase cells was increased to 71.2％ from 51.7％.Conclusion Targeted inhibition of TMPRSS2∶ ERG fusion gene expression can significantly inhibit the growth of prostate cancer cells.

关 键 词：前列腺癌 跨膜丝氨酸蛋白酶2∶ERG融合基因 RNA干扰 慢病毒 

分 类 号：R737.25[医药卫生—肿瘤]