摘要
目的研究miR-34a对乳腺癌细胞生物学功能的影响及其作用机制,为乳腺癌的诊治提供理论依据。方法实时荧光定量PCR法检测miR-34a在乳腺癌细胞系中的表达,CCK8法、Transwell实验检测miR-34a对乳腺癌细胞增殖、侵袭能力的影响,双荧光素酶法和Western blot法对靶基因Wnt1进行验证,并检测其下游基因β-catenin、CyclinD1的表达。结果 miR-34a抑制乳腺癌细胞的增殖、侵袭能力,Wnt1蛋白为miR-34a的直接作用靶点,过表达miR-34a可直接抑制Wnt1蛋白的表达,下游基因β-catenin、CyclinD1也受到抑制。结论 miR-34a直接靶向作用于Wnt1蛋白从而抑制乳腺癌细胞的增殖和侵袭。
Objective To investigate the expression profiles and potential mechanisms of microRNA-34a(miR-34a)on breast cancer cells,and to provide a theoretical basis for diagnosis and treatment of breast cancer.Methods Real-time fluorescence quantificantion PCR was used to detect the expression of miR-34a in breast cancer cells.The effect of miR-34a on the proliferation and invasion of breast cancer cells was detected by CCK8 and Transwell assay.The target gene Wnt1 was verified by dual-luciferase reporter assay and Western blot,and the expressions of its downstream genesβ-catenin and CyclinD1 were assessed.Results MiR-34a inhibited the proliferation and invasion of breast cancer cells.Wnt1 protein was the direct target of miR-34a.The expression of Wnt1 protein could be inhibited by the overexpression of miR-34a directly.Meanwhile,the expression of its downstream geneβ-catenin and CyclinD1 were inhibited as well.Conclusion MiR-34a directly targets Wnt1 protein,which suppresses the proliferation and invasion of breast cancer cells.
作者
冯刚
李良平
樊友本
刘志苏
FENG Gang;LI Liangping;FAN Youben;LIU Zhisu(Department of Thyroid Breast Surgery,the First Affiliated Hospital of China Three Gorges University/Yichang Central People′s Hospital,Yichang,Hubei 443000,China;Departmentof General Surgery,the Sixth Affiliated Hospital of Shanghai Jiaotong University,Shanghai 200000,China;Department of General Surgery,Zhongnan Hospital of Wuhan University,Wuhan,Hubei 430000,China)
出处
《重庆医学》
CAS
2019年第2期207-211,共5页
Chongqing medicine
基金
国家青年科学基金资助项目(81603345)