miR-17-92簇对慢性粒细胞白血病细胞K562增殖和凋亡的影响

Effects of miR-17-92 cluster on proliferation and apoptosis of chronic myeloid leukemia cell K562

目的探讨miR-17-92簇对慢性粒细胞白血病(CML)细胞K562增殖和凋亡的影响。方法采用RT-qPCR检测CML细胞K562中miR-17-92表达水平;采用Lipofectamin 2000转染miR-17-92 mimic,CCK-8检测转染后K562细胞增殖能力;Annexin V-FITC/PI标记,流式细胞仪检测转染后K562细胞凋亡情况,Western blotting检测转染后K562细胞中转化生长因子-β1(TGF-β1)蛋白表达水平。结果CML CD34+细胞中miR-17-92表达水平显著高于健康者CD34+细胞,差异有统计学意义(P值均<0.05)。转染miR-17-92后,K562细胞增殖能力较对照组显著升高,凋亡能力较对照组显著降低,差异有统计学意义(P值均<0.05)。转染miR-17-92后K562细胞中TGF-β1蛋白表达水平较对照组均显著升高,差异有统计学意义(P值均<0.01)。结论miR-17-92表达促进细胞的增殖能力,抑制细胞的凋亡,在CML中发挥促癌基因作用,其作用机制可能与TGF-β1信号通路的激活有关。

Objective To explore the effects of mir-17-92 on proliferation and apoptosis of chronic myelogenous leukemia(CML)cells K562.Methods The expression of miR-17-92 in CML cell K562 was detected by RT-qPCR.miR-17-92 mimic was transfected with Lipofectamin 2000,and CCK-8 was used to detect the proliferation capacity of K562 cells after transfection.Annexin V-FITC/PI labeling,flowcytometry was used to detect the apoptosis of K562 cells after transfection.The expression of TGF-β1protein in K562 cells was detected by Western blotting.Results The expression level of mir-17-92 in CML CD34+cells was significantly higher than that in normal CD34+cells,the difference was statistically significant(all P<0.05).After transfection of mir-17-92,the proliferation ability of K562 cells was significantly increased and the apoptosis ability was significantly decreased compared with the control group,the difference were statistically significant(P<0.05).After transfection of mir-17-92,the protein expression level of TGF-β1 in K562 cells was significantly higher than that in the control group,the difference was statistically significant(P<0.01).Conclusion The expression of miR-17-92 promotes the proliferation of cells,inhibits cell apoptosis,and it plays a pro-oncogene role in CML,its mechanism may be related to the activation of TGF-1 signaling pathway.