miR-577对脑胶质瘤U251细胞活力和凋亡的影响
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摘要
目的:探讨miR-577调节Wnt/β-catenin信号通路蛋白Wnt2b对脑胶质瘤U251细胞活力和凋亡的影响。方法:将U251细胞分为miR-对照组(转染miR-577阴性对照)、miR-577组(转染miR-577模拟物)、抑制对照组(转染miR-577抑制物对照)、miR-577抑制组(转染miR-577抑制物)、miR-577+Wnt2b组(转染miR-577模拟物与pc DNA-Wnt2b),采用qRT-PCR法检测转染后细胞miR-577的表达,CCK-8法及流式细胞术分别检测细胞活力和凋亡率。通过双荧光报告基因检测系统验证miR-577和Wnt2b的靶向关系。结果:miR-577组与miR-对照组相比,miR-577的表达水平升高。双荧光报告基因证实Wnt2b是miR-577的靶基因,miR-577组与miR-对照组相比Wnt2b蛋白的表达水平降低,miR-577抑制组与抑制对照组相比Wnt2b蛋白的表达水平升高(P <0. 05)。miR-577+Wnt2b组与miR-577组相比,细胞活力升高,凋亡率降低(P <0. 05)。结论:miR-577可通过下调Wnt/β-catenin信号通路Wnt2b蛋白表达抑制脑胶质瘤U251细胞活力,诱导细胞凋亡。
Aim: To investigate the effects of miR-577 on the viability and apoptosis of glioma cells U251 by regulating Wnt/β-catenin signaling pathway protein Wnt2 b. Methods: U251 cells were divided into miR-control group,miR-577 group( cells were transfected with miR-577 mimics),inhibition control group,miR-577 inhibition group,and miR-577 + Wnt2 b group( simultaneously overexpressing miR-577 and Wnt2 b). The expression of miR-577 was detected by qRT-PCR. Cell viability and apoptotic rate were detected by CCK-8 and flow cytometry,respectively. The targeting relationship between miR-577 and Wnt2 b was validated by dual fluorescence reporter gene detection system. Results: Compared with the miRcontrol group,the expression level of miR-577 increased. The dual fluorescence reporter gene confirmed that Wnt2 b was the target gene of miR-577,and the expression level of Wnt2 b was significantly decreased in miR-577 group compared with miR-control group( P < 0. 05). Compared with inhibition control group,the expression level of Wnt2 b in miR-577 inhibition group was significantly increased( P < 0. 05). Compared with miR-577 group,cell viability of miR-577 + Wnt2 b group increased,and apoptotic rate decreased( P < 0. 05). Conclusion: miR-577 could inhibit the viability of glioma cells U251 and induce apoptosis by down-regulating Wnt/β-catenin signaling pathway protein Wnt2 b.
Aim: To investigate the effects of miR-577 on the viability and apoptosis of glioma cells U251 by regulating Wnt/β-catenin signaling pathway protein Wnt2 b. Methods: U251 cells were divided into miR-control group,miR-577 group( cells were transfected with miR-577 mimics),inhibition control group,miR-577 inhibition group,and miR-577 + Wnt2 b group( simultaneously overexpressing miR-577 and Wnt2 b). The expression of miR-577 was detected by qRT-PCR. Cell viability and apoptotic rate were detected by CCK-8 and flow cytometry,respectively. The targeting relationship between miR-577 and Wnt2 b was validated by dual fluorescence reporter gene detection system. Results: Compared with the miRcontrol group,the expression level of miR-577 increased. The dual fluorescence reporter gene confirmed that Wnt2 b was the target gene of miR-577,and the expression level of Wnt2 b was significantly decreased in miR-577 group compared with miR-control group( P < 0. 05). Compared with inhibition control group,the expression level of Wnt2 b in miR-577 inhibition group was significantly increased( P < 0. 05). Compared with miR-577 group,cell viability of miR-577 + Wnt2 b group increased,and apoptotic rate decreased( P < 0. 05). Conclusion: miR-577 could inhibit the viability of glioma cells U251 and induce apoptosis by down-regulating Wnt/β-catenin signaling pathway protein Wnt2 b.
引文
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[2] LI X,CHEN W,ZENG W,et al. microRNA-137 promotes apoptosis in ovarian cancer cells via the regulation of XIAP[J]. Br J Cancer,2017,116(1):66
[3] JIANG TH,LI MF,LI QY,et al. MicroRNA-98-5p inhibits cell proliferation and induces cell apoptosis in hepatocellular carcinoma via targeting IGF2BP1[J]. Oncol Res,2017,25(7):1117
[4] YIN C,MOU Q,PAN X,et al. MiR-577 suppresses epithelial-mesenchymal transition and metastasis of breast cancer by targeting Rab25[J]. Thorac Cancer,2018,9(4):472
[5] XUE KC,HU DD,ZHAO L,et al. MiR-577 inhibits papillary thyroid carcinoma cell proliferation,migration and invasion by targeting Sph K2[J]. Eur Rev Med Pharmacol Sci,2017,21(17):3794
[6] WEI N,WEI H,ZHANG H. Long non-coding RNA ZEB1-AS1 promotes glioma cell proliferation,migration and invasion through regulating miR-577[J]. Eur Rev Med Pharmacol Sci,2018,22(10):3085
[7] WANG B,SUN L,LI J,et al. miR-577 suppresses cell proliferation and epithelial-mesenchymal transition by regulating the WNT2B mediated Wnt/β-catenin pathway in nonsmall cell lung cancer[J]. Mol Med Rep,2018,18(3):2753
[8] ZHU M,HUANG Z,ZHU D,et al. A panel of microRNA signature in serum for colorectal cancer diagnosis[J]. Oncotarget,2017,8(10):17081
[9] LI H,LIU YW,WANG H,et al. MiR-519a functions as a tumor suppressor in glioma by targeting the oncogenic STAT3 pathway[J]. J Neurooncol,2016,128(1):35
[10]何银平,叶景旺. miR-577在胃癌中的表达及对SGC-7901细胞侵袭的影响[J].医学研究杂志,2016,45(10):95
[11]WANG LY,LI B,JIANG HH,et al. Inhibition effect of miR-577 on hepatocellular carcinoma cell growth via targetingβ-catenin[J]. Asian Pac J Trop Med,2015,8(11):923
[12]张雪雅,张维溪,李昌崇. Wnt/β-catenin在气道平滑肌细胞及支气管哮喘中的作用[J].国际呼吸杂志,2014,34(23):1789
[13]WU KF,LIANG WC,FENG L,et al. H19 mediates methotrexate resistance in colorectal cancer through activating Wnt/β-catenin pathway[J]. Exp Cell Res,2017,350(2):312
[14]TAO Q,WU C,XU R,et al. Diallyl trisulfide inhibits proliferation,invasion and angiogenesis of glioma cells by inactivating Wnt/β-catenin signaling[J]. Cell Tissue Res,2017,370(3):379
[15]GAO L,CHEN B,LI J,et al. Wnt/β-catenin signaling pathway inhibits the proliferation and apoptosis of U87 glioma cells via different mechanisms[J]. PLo S One,2017,12(8):e0181346
[16]LI G,WANG YY,LIU Y,et al. miR-185-3p regulates nasopharyngeal carcinoma radioresistance by targeting WNT2B in vitro[J]. Cancer Sci,2014,105(12):1560
[17]LIU C,LI G,REN SL,et al. miR-185-3p regulates the invasion and metastasis of nasopharyngeal carcinoma by targeting WNT2B in vitro[J]. Oncol Lett,2017,13(4):2631
[18]JIANG Z,JIANG C,FANG J. Up-regulated lnc-SNHG1contributes to osteosarcoma progression through sequestration of miR-577 and activation of WNT2B/Wnt/β-catenin pathway[J]. Biochem Biophys Res Commun,2018,495(1):238