慢病毒载体介导RNA结合蛋白QKI-5过表达对前列腺癌细胞PC3增殖的抑制作用

摘要

Objective:To study RNA binding protein QKI-5 on the proliferation of androgen independent prostate cancer PC3 cell line. Methods:The expression of QKI-5 protein and mRNA was detected by using Western blot and RT-PCR respectively. Recombinant lentivirus was used to infected prostate cancer cell line PC3. MTT assays were performed to determine cell proliferation. The changes in cell cycle were detected by flow cytometer after transfected. Results:The results of western blot and RT-PCR indicated that the three prostatic carcinoma cell lines had relatively lower QKI-5 mRNA and protein levels. PC3 cells had the lowest level among the three cell lines. After infected by lentivirus that it had been verified that the QKI-5 protein over-expression was identified cells infected with recom-binant lentivirus. The MTT assay showed that QKI -5 could significantly inhibit PC3 cell proliferation(P<0. 05).Compared with the blank control and Cherry group,Lv-QKI-5 group showed an increase in G1 phase(P<0. 05). Conclusion:RNA-binding protein QKI-5 may be involved in the development of prostate cancer,lentiviral-medi-ated QKI-5 gene can inhibit the proliferation of PC3 cells.%目的：观察RNA结合蛋白Quaking-5（QKI-5）对雄激素非依赖性前列腺癌PC3细胞增殖的影响。方法：采用Western blot及逆转录-聚合酶链反应（ RT-PCR）方法检测不同前列腺癌细胞株（ PC3、LNCaP、DU145）及正常人前列腺上皮细胞（RWPE1）中QKI -5的表达水平；通过慢病毒感染并构建稳定过表达QKI-5的PC3细胞，噻唑蓝比色法（ MTT法）绘制细胞生长曲线、流式细胞仪（ FCM）检测细胞周期。结果：3株前列腺癌细胞中QKI-5的表达水平明显低于正常前列腺上皮细胞，其中PC3细胞表达水平最低，细胞生长曲线显示稳定转染QKI-5的PC3细胞增殖能力明显低于对照组（ P<0.05），FCM检测发现稳定转染QKI-5的PC3细胞同对照组比较出现了明显的G1期阻滞（ P<0.05）。结论：RNA结合蛋白QKI-5可能与前列腺癌的发生发展有着密切的联系，慢病毒介导的QKI-5基因能够抑制PC3细胞的增殖。