上调miR-126表达对白血病K562细胞增殖及凋亡的影响
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摘要
目的:探讨上调miR-126对白血病K562细胞增殖和凋亡的影响及作用机制。方法:采用脂质体转染法分别将miR-126 mimic和miR-126阴性对照(miR-126 NC)转染入K562细胞,以不做转染处理的K562细胞为空白对照。采用qRT-PCR检测转染48 h后细胞中miR-126的表达水平,转染24、48、72、96 h后MTT法检测细胞活力,转染48 h采用Annexin V-FITC/PI双染法检测细胞凋亡率,Western blot法检测细胞中PCNA及凋亡相关蛋白Bax和Bcl-2的表达。结果:与空白对照和miR-126 NC组相比,miR-126 mimic组细胞中miR-126的表达上调,细胞活力降低,细胞凋亡率升高,细胞中PCNA蛋白和Bcl-2蛋白表达下调,Bax蛋白表达上调(P均<0. 05)。结论:上调miR-126能够抑制K562细胞增殖,诱导细胞凋亡,这一效应与下调Bcl-2、促进Bax蛋白表达有关。
Aim: To investigate the effects of miR-126 up-regulation on proliferation and apoptosis of leukemia K562 cells and its mechanism. Methods: K562 cells were transfected by lipofection with miR-126 mimic or negative control( miR-126 NC). K562 cells without transfection were the blank group. The expression of miR-126 was detected by qRT-PCR.Proliferation was detected by MTT assay. Apoptosis was detected by Annexin V-FITC/PI staining. PCNA,Bax and Bcl-2 were detected by Western blot. Results: Compared with those of the blank group and miR-126 NC group,the expression of miR-126 was significantly up-regulated in miR-126 mimic group,cell viability was significantly decreased,apoptosis rate was significantly increased,PCNA and Bcl-2 protein levels were down-regulated,and Bax protein level was up-regulated( P < 0. 05). Conclusion: The up-regulation of miR-126 can inhibit the proliferation of K562 cells and induce apoptosis,which is related to down-regulating Bcl-2 protein and promoting Bax protein expression.
Aim: To investigate the effects of miR-126 up-regulation on proliferation and apoptosis of leukemia K562 cells and its mechanism. Methods: K562 cells were transfected by lipofection with miR-126 mimic or negative control( miR-126 NC). K562 cells without transfection were the blank group. The expression of miR-126 was detected by qRT-PCR.Proliferation was detected by MTT assay. Apoptosis was detected by Annexin V-FITC/PI staining. PCNA,Bax and Bcl-2 were detected by Western blot. Results: Compared with those of the blank group and miR-126 NC group,the expression of miR-126 was significantly up-regulated in miR-126 mimic group,cell viability was significantly decreased,apoptosis rate was significantly increased,PCNA and Bcl-2 protein levels were down-regulated,and Bax protein level was up-regulated( P < 0. 05). Conclusion: The up-regulation of miR-126 can inhibit the proliferation of K562 cells and induce apoptosis,which is related to down-regulating Bcl-2 protein and promoting Bax protein expression.
引文
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[3] MONDEJAR R,PEREZ C,ONAINDIA A,et al. Molecular basis of targeted therapy in T/NK-cell lymphoma/leukemia:a comprehensive genomic and immunohistochemical analysis of a panel of 33 cell lines[J]. PLo S One,2017,12(5):e0177524
[4] RUPAIMOOLE R,CALIN GA,LOPEZ-BERESTEIN G,et al. miRNA deregulation in cancer cells and the tumor microenvironment[J]. Cancer Discov,2016,6(3):235
[5]廖芬芳,吴清清,陈永恒,等.慢性粒细胞白血病干细胞microRNA的表达[J].广东医学,2016,37(19):2889
[6] TRINO S,LAMORTE D,CAIVANO AA,et al. MicroRNAs as new biomarkers for diagnosis and prognosis,and as potential therapeutic targets in acute myeloid leukemia[J].Int J Mol Sci,2018,19(2):460
[7]米旭光,李首庆,刘多,等. miR-126在乳腺癌中与VEGF表达关系及其抗肿瘤效果[J].中国免疫学杂志,2016,32(7):1000
[8] LUO P,FEI J,ZHOU J,et al. microRNA-126 suppresses PAK4 expression in ovarian cancer SKOV3 cells[J]. Oncol Lett,2015,9(5):2225
[9] ISHIHARA K,SASAKI D,TSURUDA K,et al. Impact of miR-155 and miR-126 as novel biomarkers on the assessment of disease progression and prognosis in adult T-cell leukemia[J]. Cancer Epidemiol,2012,36(6):560
[10]ZHAO CF,LI Y,ZHANG M,et al. miR-126 inhibits cell proliferation and induces cell apoptosis of hepatocellular carcinoma cells partially by targeting Sox2[J]. Hum Cell,2015,28(2):91
[11]XIANG LY,OU HH,LIU XC,et al. Loss of tumor suppressor miR-126 contributes to the development of hepatitis B virus-related hepatocellular carcinoma metastasis through the upregulation of Adam9[J]. Tumor Biol,2017,39(6):1010428317709128
[12]LIU YL,ZHOU Y,FENG X,et al. MicroRNA-126 functions as a tumor suppressor in colorectal cancer cells by targeting CXCR4 via the AKT and ERK1/2 signaling pathways[J].Int J Oncol,2014,44(1):203
[13]CHEN XM,XIONG W,LI HY. Comparison of microRNA expression profiles in K562-cells-derived microvesicles and parental cells,and analysis of their roles in leukemia[J].Oncol Lett,2016,12(6,B):4937
[14]易宗春,李扬,余春红,等. T20. 57氢醌处理K562细胞红系分化相关miRNA的表达变化[J].中国药理学与毒理学杂志,2013,27(S1):388
[15]JURIKOVAM,DANIHEL L,POLAKS,et al. Ki67,PCNA,and MCM proteins:markers of proliferation in the diagnosis of breast cancer[J]. Acta Histochem,2016,118(5):544
[16]SALVADOR-GALLEGO R,MUND M,COSENTINO KA,et al. Bax assembly into rings and arcs in apoptotic mitochondria is linked to membrane pores[J]. EMBO J,2016,35(4):389
[17]GONG C,FANG J,LI G,et al. Effects of microRNA-126 on cell proliferation,apoptosis and tumor angiogenesis via the down-regulating ERK signaling pathway by targeting EGFL7 in hepatocellular carcinoma[J]. Oncotarget,2017,8(32):52527
[18]WU XJ,ZHAO ZF,KANG XJ,et al. MicroRNA-126-3p suppresses cell proliferation by targeting PIK3R2 in Kaposi's sarcoma cells[J]. Oncotarget,2016,7(24):36614