miR-221对慢性粒细胞白血病K562细胞增殖和凋亡的影响及其作用机制  

Effects of miR-221 on cell proliferation and apoptosis of chronic myeloid leukemia K562 cells and its mechanism

作　　者：于游游 李彬[2] 闫婷 张长庚[1] YU Youyou;LI Bin;YAN Ting;ZHANG Changgeng(Department of Laboratory Medicine,Harraison International Peace Hospital,Hengshui 053000,Hebei,China;Department of Nursing,Harraison International Peace Hospital,Hengshui 053000,Hebei,China;Return Visit Center,Harraison International Peace Hospital,Hengshui 053000,Hebei,China)

机构地区：[1]哈励逊国际和平医院检验科,河北衡水053000 [2]哈励逊国际和平医院护理部,河北衡水053000 [3]哈励逊国际和平医院回访中心,河北衡水053000

出　　处：《中国肿瘤生物治疗杂志》2019年第12期1311-1317,共7页Chinese Journal of Cancer Biotherapy

基　　金：河北省指令性课题计划资助项目（No.ZL20140057）~~

摘　　要：目的:探讨下调miR-221对慢性粒细胞白血病(CML)K562细胞增殖和凋亡的影响及其相关的调控机制。方法:将K562细胞分为对照组、miRNA阴性对照(miR-NC)组、miR-221 inhibitor组、miR-221 inhibitor+阴性对照siRNA (NC siRNA)组和miR-221 inhibitor+SOCS3 siRNA组。其中,对照组细胞不进行另外处理;miR-NC组和miR-221 inhibitor组分别采用miR-NC和miR-221 inhibitor转染至细胞;miR-221 inhibitor+NC siRNA组和miR-221 inhibitor+SOCS3 siRNA组采用已稳定转染miR-221 inhibitor的细胞再分别转染NC siRNA和SOCS3 siRNA。用qPCR鉴定miR-221 inhibitor的转染效率,CCK-8法检测各组细胞的增殖活性,Annexin V-FITC/PI双染色流式术检测各组细胞的凋亡水平,WB实验检测各组细胞的SOCS3、p-JAK1、p-JAK2、p-STAT3和survivin的蛋白表达水平。结果:与对照组比较,miR-221 inhibitor组细胞内miR-221的表达显著下调(P<0.01),细胞增殖活性在转染后48、72 h时明显降低(P<0.05或P<0.01),凋亡细胞数量明显增加(P<0.01),细胞内SOCS3的表达水平明显增加(P<0.01),而p-JAK1、p-JAK2、p-STAT3和survivin的表达水平均明显降低(均P<0.01)。与miR-221 inhibitor组比较,miR-221 inhibitor+SOCS3 siRNA组细胞增殖活性在转染后24、48和72 h时明显增加(P<0.05或P<0.01),凋亡细胞数量明显减少(P<0.01),细胞内p-JAK1、p-JAK2、p-STAT3和survivin的表达水平均明显增加(均P<0.01)。结论:下调miR-221可能通过上调SOCS3表达抑制JAK-STAT3信号通路,从而抑制K562细胞增殖并促进其凋亡。Objective:To investigate the effect of down-regulation of miR-221 on cell proliferation and apoptosis of chronic myeloid leukemia(CML) K562 cells and its related regulatory mechanism.Methods:K562 cells were divided into control group,miRNA negative control(miR-NC) group,miR-221 inhibitor group,miR-221 inhibitor+negative control siRNA(NC siRNA) group and miR-221 inhibitor+SOCS3 siRNA group.The cells in the control group received no additional treatment.Cells in miR-NC group and miR-221 inhibitor group were transfected with miR-NC and miR-221 inhibitor,respectively.Cells in miR-221 inhibitor+NC siRNA group and miR-221 inhibitor+SOCS3 siRNA group were transfected with NC siRNA and SOCS3 siRNA,respectively,on the basis of successful transfection with miR-221 inhibitor.The transfection efficiency of miR-221 inhibitor was identified by qPCR.Cell viability in each group was measured by CCK-8 assay.Apoptosis in each group was detected by Annexin V-FITC/PI staining using a flow cytometry.The protein expressions of SOCS3,p-JAK1,p-JAK2,p-STAT3 and survivin in each group were detected by WB.Results:Compared with the control group,miR-221 expression was significantly down-regulated in miR-221 inhibitor group(P<0.01),cell viability was significantly reduced at 48 and 72 h after transfection(P<0.05 or P<0.01),the number of apoptotic cells was significantly increased(P<0.01),the expression of SOCS3 was significantly increased(P<0.01) and the expression levels of p-JAK1,p-JAK2,p-STAT3 and survivin were significantly reduced(all P<0.01).Compared with miR-221 inhibitor group,cell viability was significantly increased at 24,48 and 72 h after transfection(P<0.05 or P<0.01),the number of apoptotic cells was significantly decreased(P<0.01) and the expression levels of p-JAK1,p-JAK2,p-STAT3 and survivin were significantly increased in miR-221 inhibitor+SOCS3 siRNA group(all P<0.01).Conclusion:Down-regulation of miR-221 inhibits proliferation and promotes apoptosis of K562 cells,the mechanism of which may be related with up-regulating SO

关 键 词：MIR-221 慢性粒细胞白血病 凋亡 增殖 JAK-STAT信号通路 

分 类 号：R733.72[医药卫生—肿瘤] R392.12[医药卫生—临床医学]