Bloom解旋酶基因RNA干扰载体对前列腺癌PC3细胞的抑制作用  

Inhibitory effect of Bloom helicase gene targeted RNA interference vector against prostate cancer PC3 cells

作　　者：罗霂榃 许厚强[2] 刘忠伟[2,3] 段志强[2] 赵佳福[1,2] 吴萍[1,2] 陈福[1,2] 

机构地区：[1]贵州大学生命科学学院,贵阳550025 [2]贵州大学动物科学学院高原山地动物遗传育种与繁殖教育部重点实验室,贵阳550025 [3]贵州大学农学院,贵阳550025

出　　处：《第二军医大学学报》2016年第6期699-703,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(31361406);贵州省国际合作项目[黔科合外G字(2011)7008号]

摘　　要：目的采用RNA干扰(RNAi)技术抑制前列腺癌PC3细胞系中Bloom解旋酶基因的表达,探讨Bloom解旋酶基因表达下调后对PC3细胞的抑制作用。方法使用实验室前期成功构建的两条针对于Bloom解旋酶基因的RNAi载体shRNA-1和shRNA-2转染前列腺癌PC3细胞,以未转染RNAi载体为对照组,分别在转染24、48、72h后通过MTT法检测细胞增殖情况,转染48h后通过Transwell小室法检验细胞侵袭、迁移能力,细胞划痕实验检测细胞迁移情况,Hoechst/PI双染法检测细胞凋亡情况。结果转染RNAi载体后,与未转染对照组相比,各实验时间点的转染组细胞增殖率降低(P<0.05),Transwell细胞侵袭和迁移实验中穿过室膜的细胞数与对照组比均减少(侵袭:119±24、118±30vs 227±38;迁移:122±13、121±47vs 277±32,P<0.05),划痕愈合率与划痕迁移距离均降低,48h时差异有统计学意义(P<0.05),而且细胞凋亡明显增加。结论以RNAi载体干扰Bloom解旋酶基因的表达可抑制前列腺癌PC3细胞的增殖、迁移和侵袭能力并促进其凋亡,为前列腺癌的靶向基因治疗提供了依据。Objective To explore the inhibitory effect of down-regulating Bloom helicase gene using RNA interference （RNAi） vectors on prostate cancer PC3 cells. Methods The BLM-RNAi vectors constructed previously in our lab were transfected into prostate cancer PC3 cells, and the proliferation ability of PC3 cells was detected with MTT at 24 h, 48 h and 72 h after transfection of recombinant vectors. At 48 h, the invasion and migration ability was detected with Transwell chamber assay, migration distance was examined with cell scratch test, and cell apoptosis was detected with Hoechst/PI. Results The proliferation ability of cells after transfection was significantly weaker than that in the blank control group （P〈0. 05）, the number of cells across Transwell membrane after transfection was significantly less than that in the blank control group （invasion ability： 119 ± 24, 118 ± 30 vs 227±38, P〈0. 05; migration ability; 122±13, 121±47 vs 2774-32, P〈0. 05）, cell scratch test also showed that the wound healig rate and cell migration distance in the experimental group were significantly lower than those in the blank control group （P〈0. 05）, and cell apoptosis in the experimental group was more notable compared with the blank control group. Conclusion RNAi vectors targeting Bloom helicase gene can inhibit the proliferation, migration and invasion ability of prostate cancer PC3 cells, and increase the cell apoptosis, which may cast new lights on the gene therapy of prostate cancer.

关 键 词：RNA干扰 前列腺肿瘤 Bloom解旋酶 细胞增殖 细胞迁移 细胞侵袭 细胞凋亡 

分 类 号：R737.25[医药卫生—肿瘤]