MicroRNA-193b对K562细胞bcr/abl表达的影响

MicroRNA-193b down-regulates bcr/abl in K562 cells

目的用Ad-pre-miRNA-193b重组腺病毒感染K562细胞,探讨miR-193b在细胞中过表达对bcr/abl表达的影响。方法将前期构建的Ad-pre-miRNA-193b重组腺病毒感染K562细胞后,采用荧光定量PCR(qRT-PCR)检测miR-193b的表达;qRT-PCR检测bcr/abl融合基因的表达;Western blot检测BCR/ABL融合蛋白的表达;CCK-8检测K562细胞的增殖情况,流式细胞术(FCM)检测K562细胞凋亡率。结果与对照比较,K562细胞感染Ad-pre-miRNA-193b重组腺病毒后,miR-193b表达明显升高;bcr/abl融合基因及BCR/ABL融合蛋白表达明显下调;K562细胞增殖能力明显降低(P<0.05),凋亡水平明显升高(P<0.05)。结论 miR-193b可下调bcr/abl表达,抑制K562细胞的生物学效应,过表达miR-193b能作为治疗CML的有效手段。

Objective To determine the effect of miR-193 b over-expression on the expression of bcr/abl in chronic myelocytic leukemia K562 cells by transfection of recombinant adenovirus of Ad-pre-microRNA-193 b. Methods After K562 cells were infected by Ad-pre-microRNA-193 b virus,the expression of miR-193 b was detected by real-time PCR. The expression of bcr/abl at mRNA and protein levels was detected by qRT-PCR and Western blotting. Cell proliferation was measured by CCK-8 assay,and cell apoptosis was tested by flow cytometry. Results Compared with control cells, the level of microRNA-193 b was significantly up-regulated in K562 cells after Ad-pre-microRNA-193 b infection. The expression of bcr/abl was significantly decreased,and BCR/ABL fusion protein was decreased too. The virus infection resulted in significant inhibition in cell proliferation and obvious improvement in apoptotic rate( both P < 0. 05).Conclusion miR-193 b down-regulates the expression of bcr/abl and inhibits the biological effects of K562 cells. The over-expression of miR-193 b may be an effective target in the treatment of chronic myelogenous leukemia.