下调miR-125b以抑制白血病K562细胞的增殖

Inhibitory Effect of miR-125b Down-Regulation on Proliferation of Leukemia Cell K562

目的:探讨下调mi R-125b对白血病K562细胞的增殖抑制作用及相关机制。方法:利用LipofectamineTM2000脂质体将mi R-125b inhibitor和mi R-125b NC转染于白血病K562细胞中,应用RT-PCR法检测mi R-125b表达,MTT法检测细胞活力,琼脂糖形成实验检测细胞集落形成能力,流式细胞术检测细胞周期,Western blot检测细胞中BCL-2、BCL-2同源拮抗物1(Bak1)、p53及p53正向细胞凋亡调控因子(Puma)表达。结果:与mi R-125b NC比较,mi R-125b inhibitor组mi R-125b表达下调(P<0.01),细胞活力及细胞集落形成能力降低(P<0.01),细胞周期阻滞在G1期(P<0.01),BCL-2表达下调(P<0.01),BAK1、p53和Puma表达上调(P<0.01)。结论:mi R-125b表达量下调能显著地抑制白血病细胞K562的增殖,这一效应与下调BCL-2表达、上调BAK1、p53及Puma表达有关。

Objective： To explore inhibitory effect of miR - 125b down - regulation on proliferation of leukemia cell K562. Methods： miR-125b inhibitor and miR-125b NC were transfected into K562 cells by liposome LipofectamineTM 2000, the cell viability was measured by MTT assay, cell cloning ability was detected by agarose cloning assay, cell cycle was measured by flow cytometry. The expression of BCL-2, BCL-2 homology antagonist/liller 1 （ BAK1 ）, p53 and p53 up-regulated modulator of apoptosis （Puma） was measured by Western blot. Results： Compared with miR- 125b NC, the expression of miR-125b was down-regulated （P 〈 0. 01 ）, the cell viability and cell cloning ability were reduced （ P 〈 0.01 ）, the cell cycle was arrested in the G1 phase （ P 〈 0.01 ） , the expression of BCL-2 were down- regulated （P 〈 0.01 ）, the expression of BAK1, p53 and Puma were up-regulated in miR-125b inhibitor group （P 〈 0.01）. Conclusion： Down-regulation of miR-125b can inhibit K562 cell proliferation via down-regulating the expressions of BCL-2 and up-regulating the expression of BAK1, p53 and Puma.