摘要
目的研究microRNA-616(miR-616)在肝细胞性肝癌(hepatocellular carcinoma,HCC)组织中的表达,探讨其与HCC上皮-间质转化(epithelial-mesenchymal transition,EMT)的关系及临床意义。方法用实时定量PCR检测40例HCC组织及癌旁组织及人正常肝细胞株LO2和6种HCC细胞株Hep3B、Hep G2、Huh7、MHCC97L、HCCLM3和MHCC97H中miR-616表达。经细胞转染分别在Hep3B细胞中过表达miR-616及在MHCC97H细胞沉默miR-616,通过Western blot、荧光素酶报告检测方法了解相应HCC细胞中miR-616、E-cadherin及vimentin的表达情况。结果与对应的癌旁组织相比,miR-616在肿瘤组织中的表达水平显著升高(0.031±0.043 vs 0.12±0.078,P<0.01)。所有6种HCC细胞株的miR-616表达水平均显著高于人正常肝细胞株LO2(P<0.01)。在Hep3B细胞中过表达miR-616,可降低E-cadherin表达水平(P<0.01),增加vimentin的表达(P<0.01)。在MHCC97H细胞中抑制miR-616的表达,可增加E-cadherin的表达水平(P<0.01),降低vimentin表达(P<0.01)。结论 miR-616可作为HCC一种有价值的临床生物标志物,靶向干预miR-616,可影响HCC的EMT。
Objective To investigate the expression of microRNA-616（ miR-616） in hepatocellular carcinoma（ HCC）,and to explore its relationship with epithelial-mesenchymal transition（ EMT） of HCC and its clinical significance. Methods The miR-616 expression was detected by real-time quantitative PCR method in specimens of primary cancer and their adjacent non-cancerous tissues from40 HCC patients and in HCC cells with different differentiation degree. The miR-616 was transfected into Hep3 B cells and miR-616 inhibitor was transfected into MHCC97 H cells,and then the expression of miR-616 was detected by luciferase reporter assay and the levels of E-cadherin,vimentin were detected by Western blot. Results The expression level of miR-616 in tumor tissues was significantly higher than that of the corresponding adjacent cancer tissues（ 0. 12 ± 0. 078 vs 0. 031 ± 0. 043,P〈0. 01）. The expression level of miR-616 was significantly higher in 6 kinds of HCC cells than in normal human liver cell line L02（ P〈0. 01）. The expression level of E-cadherin was reduced and vimentin was elevated in Hep3 B cells with miR-616 overexpression vector transfected（ P〈0. 01）. The expression level of E-cadherin was elevated and vimentin was reduced in MHCC97 H cells transfected with miR-616 inhibitor（ P〈0. 01）.Conclusion MiR-616 can be used as a valuable biomarker for HCC,and it can affect the EMT of HCC by targeting miR-616.
作者
张立
张迪
李孝彬
王炜
孙学军
李杰
李军辉
ZHANG Li ZHANG Di LI Xiaobin WANG Wei SUN Xuejun LI Jie LI Junhui(Department of General Surgery, Second Affiliated Hospital of Xi＇ an Jiaotong University, Xi＇ an 710004, China Department of General Surgery, First Affiliated Hospital of Xi＇ an Jiaotong University)
出处
《山西医科大学学报》
CAS
2016年第9期795-799,共5页
Journal of Shanxi Medical University
基金
陕西省社会发展科技攻关基金资助项目(2016SF-137)
