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作 者:王开琼[1] 邢贻雷[1] 乔欣 李仕总 宫东伟 余智威 吴奕强[1] WANG Kaiqiong;XING Yilei;QIAO Xin;LI Sizong;GONG Dongwei;YU Zhiwei;WU Yiqiang(Department of Biliary and Pancreatic Surgery,Hainan General Hospital,Haikou 570311,China)
机构地区:[1]海南省人民医院胆胰外科
出 处:《肿瘤防治研究》2019年第10期879-883,共5页Cancer Research on Prevention and Treatment
摘 要:目的探讨miR-144对胰腺癌SW1990细胞增殖、迁移、侵袭及磷脂酰肌醇-3激酶(PI3K)通路的影响。方法采用实时荧光定量PCR(RT-qPCR)检测人正常胰腺上皮细胞系HPDE与胰腺癌SW1990细胞中miR-144表达水平。将miR-144阴性对照质粒、miR-144 mimic质粒分别转染至SW1990细胞,分别命名为阴性转染组、miR-144过表达组,未经处理的细胞命名为空白对照组。采用RT-qPCR法检测各组miR-144表达水平;CCK-8法检测细胞增殖情况;平板细胞克隆形成实验检测菌落形成;Transwell检测细胞迁移及侵袭能力;Western blot法检测PI3K通路中PI3K及其磷酸化蛋白(p-PI3K)、丝氨酸-苏氨酸蛋白激酶(Akt)及其磷酸化蛋白(p-Akt)的表达水平。结果SW1990细胞中miR-144的表达水平显著低于HPDE细胞(P<0.05);miR-144过表达组中miR-144表达水平显著高于空白对照组和阴性转染组(P<0.05)、细胞增殖率、菌落形成、迁移及侵袭数量和p-PI3K、p-Akt蛋白表达水平均显著低于空白对照组、阴性转染组(均P<0.05)。结论miR-144在胰腺癌细胞SW1990中呈低表达,上调miR-144表达水平有效抑制胰腺癌细胞增殖、迁移及侵袭,其机制可能与PI3K通路抑制有关。Objective To investigate the effects of microRNA-144(miR-144)on the proliferation,migration,invasion and phosphoinositide-3 kinase(PI3K)pathway of pancreatic cancer SW1990 cells.Methods The expression of miR-144 in human normal pancreatic epithelial cell line HPDE and pancreatic cancer cells SW1990 were detected by RT-qPCR.SW1990 cells were transfected with miR-144 negative control plasmid(negative transfection group)and miR-144 mimic plasmid(miR-144 overexpression group),and untreated cells were taken as blank control group.The expression level of miR-144 in each group was detected by RTqPCR.CCK-8 assay was used to detect cell proliferation.Plate clone formation assay was used to detect colony formation.Transwell assay was used to detect cell migration and invasion.The expression levels of PI3K and phosphorylated protein(p-PI3K),serine-threonine protein kinase(Akt)and phosphorylated protein(p-Akt)proteins in PI3K pathway were detected by Western blot.Results The expression level of miR-144 in SW1990 cells was significantly lower than that in HPDE cells(P<0.05).The expression level of miR-144 in miR-144 overexpression group was significantly higher than those in blank control group and negative transfection group(P<0.05).Cells proliferation rate,colony formation,the number of migration and invasion cells and the expression levels of p-PI3K and p-Akt proteins in the overexpression group were significantly lower than those in the blank control group and negative transfection group(all P<0.05).Conclusion miR-144 is lowly expressed in pancreatic cancer cells SW1990.The up-regulation of miR-144 expression could effectively inhibit the proliferation,migration and invasion of pancreatic cancer cells,which may be related to the inhibition of PI3K pathway.
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