摘要
目的探讨肝细胞生长因子(HGF)及受体(c-MET)信号通路在乳腺癌上皮-间充质转化(EMT)及细胞增殖侵袭中的作用。方法利用c-MET激酶抑制剂PHA-665752体外处理人乳腺癌MCF-7细胞,分为研究组(加入抑制剂)和对照组(加入等量二甲基亚砜)。使用CCK-8法检测乳腺癌MCF-7细胞的增殖能力,Transwell实验检测细胞的迁移和侵袭能力,蛋白质印迹(Western blot)法检测细胞EMT相关蛋白上皮钙黏蛋白(E-cad)、锌指转录因子(Snail)及神经型钙黏蛋白(N-cad)表达水平,实时荧光定量聚合酶链反应(qRT-PCR)法检测c-MET、E-cad mRNA相对表达量。结果c-MET激酶抑制剂剂量依赖性地抑制乳腺癌MCF-7细胞的增殖能力。0.5μmol/L PHA-665752研究组MCF-7细胞处理24 h后侵袭细胞数为(24.33±1.53),少于对照组的(93.00±5.57),差异有统计学意义(P<0.05)。处理24 h时,0.5μmol/L PHA-665752组乳腺癌MCF-7细胞c-MET mRNA相对表达量低于对照组,E-cad mRNA相对表达量高于对照组,差异均有统计学意义(P<0.05)。0.5μmol/L PHA-665752组MCF-7细胞E-cad蛋白表达水平高于对照组,N-cad、Snail蛋白表达水平均低于对照组,差异均有统计学意义(P<0.05)。结论c-MET激酶抑制剂可能通过抑制乳腺癌EMT,进而抑制肿瘤细胞的侵袭和转移。
Objective To investigate the action of hepatocyte growth factor(HGF)and receptor(c-MET)signaling pathway in breast cancer epithelial-mesenchymal transition(EMT),cell proliferation and cell invasion.Method PHA-665752,a c-MET kinase specific inhibitor,was used to treat human breast cancer MCF-7 cells in vitro,and then they were divided into a study group(cultured with inhibitor)and a control group(cultured with equal amount of dimethyl sulfoxide).The proliferation ability of human breast cancer cells was detected by CCK-8 assay.Cell migration and invasion ability was detected by Transwell assay.The cellular EMT-related protein E-cadherin(E-cad)and Zinc finger transcription factor(Snail)and N-cadherin(N-cad)expression levels were measured by the Western blot method,and the relative expression of c-MET and E-cad mRNA were measured by real-time quantitative polymerase chain reaction(qRT-PCR)method.Result c-MET kinase inhibitor inhibited the proliferation ability of breast cancer MCF-7 cells in a dose-dependent manner.The number of migratory cells in the study group where MCF-7 cells were treated with 0.5μmol/L PHA-665752 for 24 h was(24.33±1.53),which was less than that of the control group whose value was(93.00±5.57),and the difference was statistically significant(P<0.05).After treated for 24 h,the relative expression of c-MET mRNA in breast cancer MCF-7 cells in the study group(0.5μmol/L PHA-665752)was lower than that in the control group while the relative expression of E-cad mRNA was higher than that in the control group.The differences were statistically significant(P<0.05).The E-cad protein expression level of MCF-7 cells in the study group was higher than that of the control group,and the N-cad and Snail protein expression levels were lower than those of the control group,and the differences were statistically significant(P<0.05).Conclusion c-MET kinase inhibitors may inhibit EMT of breast cancer,and thereby inhibiting invasion and metastasis of tumor cells.
作者
刘海旺
张宏旭
刘燃
郝美玲
李春辉
LIU Haiwang;ZHANG Hongxu;LIU Ran;HAO Meiling;LI Chunhui(Department of Pathology,Affiliated Hospital of Chengde Medical University,Chengde 067000,Hebei,China;Department of Breast Surgery,Affiliated Hospital of Chengde Medical University,Chengde 067000,Hebei,China;Department of Anesthesiology,Affiliated Hospital of Chengde Medical University,Chengde 067000,Hebei,China)
出处
《癌症进展》
2020年第20期2080-2083,共4页
Oncology Progress
基金
承德市科学技术研究与发展计划项目(201701A080)。
