Survivin-siRNA真核重组质粒的构建及对前列腺癌DU145细胞增殖抑制作用  

Construction of Survivin siRNA Expression Vector and the Inhibition Effects on Prostate Neoplasm DU145 Cell Growth

作　　者：刘艳波[1] 耿艳 葛贺[1] 芦丽莉[1] 

机构地区：[1]北华大学基础医学院,吉林吉林132013 [2]吉林市丰满区疾病预防控制中心,吉林吉林132013

出　　处：《北华大学学报（自然科学版）》2010年第2期129-133,共5页Journal of Beihua University（Natural Science）

基　　金：国际科技合作重点项目(2004DFB02000)

摘　　要：目的应用RNA干扰技术（RNAi），构建针对凋亡抑制因子survivin的siRNA，并检测其对DU145细胞增殖抑制的影响．方法构建重组质粒shRNA-sur1和shRNA-sur2并分别转染DU145细胞株，通过MTT法和流式细胞术观察其对细胞的增殖抑制作用及凋亡的影响．结果成功构建针对survivin干扰的两种真核表达质粒，转染72h后，两个实验组细胞的生长速度分别是脂质体组的（37．2％±5．8％）（n=3）和（43．5％±7．2％）（n=3）．流式细胞术发现：实验组细胞的G1期细胞比例明显高于对照组，而G2期和S期细胞比例减少，细胞出现凋亡，与脂质体组比较，两实验组细胞的凋亡率分别为（21．70％±6．45％），（14．835±5．65％）．结论成功构建针对survivin siRNA的真核重组质粒，两种质粒体外均可抑制DU145增殖并诱导其凋亡，为survivin介导的肿瘤基因沉默提供良好的基础．Objective To construct survivin siRNA expression vector by using RNA inference technique and make clear the role of survivin specific siRNA on DU145 proliferation inhibition. Method The recombinant plasmid shRNA-sur1 and shRNA-sur2 were constructed and infected into prostate carcinoma DU145 cells. Proliferation abilities were measured by MTT, and the cell cycle and apoptosis were assayed by FCM. Results Survivin specific siRNA expression vectors were successfully constructed and identified. After 72 h transfection,the proliferation rate of cells in shRNA-surl and shRNA-sur2 groups was inhibited according to MTT, about 37.2% ± 5.8% （ n = 3 ） and 43.5%±7.2% （ n = 3 ） of that in lipofectin group. Cell numbers of G1 phase in two siRNA groups were significantly higher than those in controls,while cells of G2 phase and S phase were much lower. Cell apoptosis was found in both siRNA groups and the apoptotic rate were 21.70% ±6.45% and 14. 835 ± 5.65% respectively compared with lipofectin. Conclusion Recombinant plasmids expressing siRNA of human survivin gene were constructed successfully. Two kinds of plasmids could inhibit DU145 cells proliferation and induce their apoptosis in vitro and shed light on a new strategy in gene silence therapy targeting survivin.

关 键 词：SIRNA DU145细胞株 SURVIVIN基因 前列腺癌 

分 类 号：R737.25[医药卫生—肿瘤]