反义微小RNA-21对人膀胱癌细胞增殖和凋亡的影响

Effect of antisense miRNA-21 on proliferation and apoptosis of bladder cancer cells

目的:探讨微小RNA-21(miRNA-21)反义寡核苷酸(ASOND)对人膀胱癌细胞株T24增殖和凋亡的影响,为膀胱癌的治疗提供理论依据。方法:将脂质体介导的反义寡核苷酸(AS-miRNA-21)转染T24细胞,设转染无义序列组、对照组和AS-miRNA-21转染组,采用蛋白印迹(Western blotting)检测转染细胞miRNA-21表达水平,噻唑蓝(MTT)比色法和流式细胞仪(FCM)分别检测转染细胞的增殖和凋亡情况,转染无义序列组、对照组分别与AS-miRNA-21转染组比较。结果:经过转染AS-miRNA-21的T24细胞miRNA-21表达水平下降,细胞增殖能力受抑。AS-miRNA-21组T24细胞增殖率为53.6%,转染无义序列组T24细胞增殖率为92.5%,对照组为100%;相对应的凋亡率分别为13.8%、1.9%和2.7%。上述3组间T24细胞的增殖率、凋亡率比较差异均有显著性(P<0.01)。结论:反义miRNA-21对人膀胱癌细胞T24具有抑制增殖和促进凋亡的作用,抑制miRNA-21表达有望成为膀胱癌基因治疗的有效方法。

Objective To investigate the effect of miRNA-21 antisense oligonucleotide（ASOND） on the proliferation and apoptosis of bladder cancer cell line T24,and provide theoretical basis for treatment of bladder cancer. Methods AS-miRNA-21 mediated by liposome was transfected into T24 cells. As-miRNA-21 group,transfection nonsense series group and control group were set up.The protein expression level of miRNA-21 of the transfected cells was detected by Western blotting,and the proliferation and apoptosis of the transfected cells were measured by MTT and FCM.Results The miRNA-21 expression level of the tumor cells transfected with AS-miRNA-21 was decreased,the cell proliferation was inhibited.The proliferation rates of T24 cells in AS-miRNA-21 group,transfection nonsense series group and control group were 53.6%,92.5%,and 100%,respectively; the relative apoptotic rates were 13.8%,1.9% and 2.7%.There were significant differences of the proliferation rate and apoptotic rates between the three groups（P0.01）.Conclusion Antisense miRNA-21 has the action of proliferation inhibition and apoptosis enhancement on human bladder cancer cell T24. Inhibiting the expression of miRNA-21 may become a prospective gene therapy for bladder cancer.