miR-29b通过PI3K/AKT信号通路抑制白血病细胞的增殖

Mechanism and effect of miR-29b on proliferation and apoptosis in leukemia cell

目的探讨miR-29b对K562白血病细胞增殖与凋亡的影响及相关机制。方法利用Lipofectamine^(TM)2000脂质体将miR-29b mimics,miR-29b NC转入白血病细胞K562中,实时荧光定量PCR法检测miR-29b表达,MTT法检测细胞活力,EdU染色法检测细胞增殖情况,流式细胞术检测细胞周期,Western印迹检测细胞中B淋巴细胞瘤(Bcl)-2、Bcl-2相关X蛋白(Bax),髓样细胞白血病(MCL)-1,细胞周期蛋白依赖性激酶(CDK)4,CDK6蛋白表达及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)激活情况。结果与miR-29b NC组比较,miR-29b mimics组miR-29b表达上调(P<0.01),细胞活力及细胞增殖率降低(P<0.01),细胞周期阻滞在G1期(P<0.01),Bcl-2,MCL-1,CDK4,CDK6,PI3K及p-AKT表达量均明显下调(P<0.01),Bax表达量明显上调(P<0.01)。结论 miR-29b mimics能显著抑制白血病细胞K562增殖、诱导细胞凋亡,可能与阻断PI3K/AKT信号通路有关。

Objective To explore mechanism and effect of miR-29b on proliferation and apoptosis in leukemia cell K562.Methods miR-29b mimics and miR-29b NC were transfected into K562 cell by liposome Lipofectamine TM2000. The expression of miR-29b was detected by Quantitative Real-time PCR. Cell viability was measured by MTT assay. Cell proliferation was measured by Ed U staining. Cell cycle was measured by flow cytometry. The expressions of B-cell lymphoma-2（ Bcl-2）,Bcl-2 associated X protein（ Bax）,myeloid cell leukemia（ MCL）-1,cell cyclin-dependent kinase（ CDK） 4 and CDK6,the activation of phosphatidylinositol 3 kinase（ PI3K）/protein kinase B（ AKT） signal pathway were measured by Western blot.Results Compared with miR-29b NC,the expression of miR-29 b was up-regulated（ P0.01）,cell viability and cell proliferation were reduced（ P0.01）,cell cycle was made arrested in the G1 phase（ P0.01）,the expressions of Bcl-2,MCL-1,CDK4,CDK6,PI3 K and p-AKT were down-regulated（ P 0. 01）,the expression of Bax was up-regulated in miR-29b mimics group（ P0.01）. Conclusions miR-29b mimics could inhibit K562 cell proliferation and induce cell apoptosis via blocking PI3K/AKT signal pathway.