【摘要】 目的：探讨miR-217对急性髓系白血病细胞增殖及阿霉素敏感性的影响。方法：构建mimic NC和mi R-217 mimic载体转染至HL-60细胞中，q PCR检测转染效率。不同浓度的阿霉素处理细胞24和48 h,CCK-8法检测阿霉素化学敏感性并筛选阿霉素的最佳处理浓度和时间。将细胞分为对照、mimic NC、mi R-217 mimic、阿霉素和mi R-217 mimic+阿霉素共5组，流式细胞术检测细胞凋亡，qPCR检测miR-217、PI3K和Akt3的表达，Western blot检测PI3K/Akt通路蛋白PI3K、Akt3和凋亡蛋白Bcl-2、Bax的表达，双荧光素酶验证mi R-217和Akt3的关系。结果：mi R-217 mimic能够增强HL-60细胞对阿霉素的敏感性，阿霉素的最佳处理浓度和时间为160 ng/ml、48 h。与对照组相比，mi R-217 mimic组和阿霉素组细胞凋亡率、mi R-217和Bax蛋白水平显著升高（P<0.01），而Bcl-2蛋白和PI3K、Akt3 m RNA和蛋白水平显著降低（P<0.01）；与阿霉素组相比，mi R-217 mimic+阿霉素组细胞凋亡率、mi R-217和Bax蛋白水平显著升高（P<0.01）,Bcl-2蛋白和PI3K、Akt3 m RNA和蛋白水平显著降低（P<0.01）。双荧光素酶实验表明mi R-217与Akt3之间存在靶向调控关系。结论：mi R-217靶向Akt3调控PI3K/Akt通路，抑制细胞增殖，促进细胞凋亡并增强阿霉素对急性髓系白血病细胞的敏感性。更多还原

【Abstract】 Objective: To investigate the effects of mi R-217 on proliferation and adriamycin sensitivity of acute myeloid leukemia(AML) cells. Methods: The mimic NC and mi R-217 mimic vectors were constructed and transfected into HL-60 cells, and transfection efficiency was detected by q PCR. The cells were treated with different concentrations of adriamycin for 24 h and 48 h. CCK-8 assay was used to detect the chemical sensitivity of adriamycin and screen the optimal concentration and time of adriamycin treatment. Cells were divided into control group, mimic NC group, miR-217 mimic group, adriamycin group and mi R-217 mimic+adriamycin group. Apoptosis was detected by flow cytometry, and the expressions of miR-217, PI3K and Akt3 were detected by qPCR. Western blot was used to detect the expression of PI3K/Akt pathway proteins PI3K, Akt3 and apoptosis proteins Bcl-2, Bax, and double luciferase was used to verify the relationship between miR-217 and Akt3. Results: MiR-217 mimic could enhance the sensitivity of HL-60 cells to adriamycin. The optimal concentration and treatment time of adriamycin were 160 ng/ml and 48 h, respectively. Compared with control group, apoptosis rate, mi R-217 and Bax protein levels were significantly increased in miR-217 mimic and adriamycin groups(P<0.01), while Bcl-2 protein, PI3K, Akt3 mRNA and protein levels were significantly decreased(P<0.01). Compared with adriamycin group, apoptosis rate, miR-217 and Bax protein levels were significantly increased in miR-217 mimic+adriamycin group(P<0.01), while Bcl-2 protein, PI3K, Akt3 mRNA and protein levels were significantly decreased (P<0.01). Dual luciferase assay showed that there was a targeted regulatory relationship between miR-217 and Akt3. Conclusion: MiR-217 regulates the PI3K/Akt pathway targeting Akt3, inhibits cell proliferation, promotes cell apoptosis and enhances the sensitivity of adriamycin to AML cells.更多还原