miR-148b在急性髓系白血病患者及细胞系中的表达与生物学功能

Preliminary study on expression and biological functions of miR-148b in acute myeloid leukemia patients and cell lines

目的:检测miR-148b在急性髓系白血病患者和细胞系中的表达水平。初步研究miR-148b在急性髓系白血病中的生物学功能。方法:收集急性髓系白血病患者56例,非恶性血液病对照组20例,提取骨髓有核细胞;培养U937、Kasumi-1、THP-1三种AML细胞系。RT-PCR检测miR-148b的表达水平。分别对U937细胞系及Kasumi-1细胞系转染miR-148b mimics和inhibitor。CCK-8法检测U937细胞及Kasumi-1细胞增殖情况,Annexin V/PI流式检测细胞凋亡,流式细胞术检测细胞周期。结果:miR-148b在急性髓系白血病患者及细胞系中较非恶性血液病对照组表达明显下降。U937细胞转染miR-148b mimics后,增殖受抑制,细胞凋亡增加,细胞阻滞在G_0-G_1期;Kasumi-1细胞转染miR-148b inhibitor后,增殖增加,细胞凋亡减少,G_0-G_1期细胞减少。结论:miR-148b在急性髓系白血病患者及细胞系中的表达水平均较正常对照显著降低。miR-148b在急性髓系白血病中起抑癌基因的作用,过表达miR-148b可以抑制细胞增殖,促进凋亡。

Objective：To detect the expression level of miR - 148b in acute myeloid leukemia and AML cell lines. A preliminary study on the biological function of miR - 148b in AML was also conducted. Methods：56 patients with AML and 20 cases with non - malignant blood disease were collected. AML cell lines U937,THP - 1 and Kasumi- 1 were cultured. Expression level of miR - 148b was detected by RT - PCR. miR - 148b mimics/ inhibitor respec-tively transfected U937 cell lines and Kasumi - 1 cell lines. U937 cell and Kasumi - 1 cell proliferation were detected by CCK - 8 method. Flow cytometry was used to detect cell apoptosis and cell cycle. Results：miR - 148b was signifi-cantly decreased in AML patients and AML cell lines compared with non - malignant hematologic disease control group. miR - 148b mimics were transfected into U937 cell line. U937 cells were blocked in G0 - G1 phase,prolifera-tion was inhibited and apoptosis was increased after transfection. miR - 148b inhibitor were transfected into Kasumi -1 cell lines,showing that cells in G0 - G1 were reduced,proliferation increased and apoptosis was decreased. Conclu-sion：Expression of miR - 148b was significantly lower in AML patients and cell lines than normal control. miR - 148b plays a role of tumor suppressor genes in AML. Over expression of miR - 148b can inhibit cell proliferation,promote apoptosis.