MiR-486-5p在肝癌中的表达及其对肝癌细胞生长的影响  

Expression of miR-486-5p in hepatocllular cancer and its effect on the growth of HCC cells

作　　者：黄馨萍 侯晋[2] 沈筱芸[1] 陆海娟[1] 罗小玲[1] 

机构地区：[1]广西医科大学附属肿瘤医院实验研究部,南宁530021 [2]第二军医大学医学免疫学国家重点实验室,上海200433

出　　处：《现代免疫学》2014年第1期7-13,共7页Current Immunology

基　　金：第二军医大学免疫学研究所暨医学免疫学国家重点实验室开放课题

摘　　要：本研究旨在探讨肝癌中miR-486-5p的表达及其对肝癌细胞生长的影响。首先,通过qRT-PCR对40例肝癌患者的肝癌组织及其癌旁组织、以及肝癌细胞系(QGY-7701、QGY-7703、BEL-7404、SMMC-7721、Huh7、HepG2和PCL/PRF/5)进行miR-486-5p表达水平的验证;然后,合成miR-486-5p模拟物及其阴性对照物NC,分别转入上述七种肝癌细胞,采用CCK8试剂检测miR-486-5p对肝癌细胞生长的影响;最后,采用流式细胞术检测miR-486-5p对肝癌细胞周期和细胞凋亡的影响。结果显示,与miRNA第二代高通量大规模平行测序(miRNomes MPSS)结果一致,miR-486-5p在肝癌组织和肝癌细胞中显著低表达(P<0.05)。CCK8检测结果表明,转染miR-486-5p模拟物能有效、稳定的抑制上述七种肝癌细胞的生长(P<0.05)。流式细胞仪检测发现,过表达miR-486-5p能促进肝癌细胞凋亡(QGY-7703细胞,P<0.001;QGY-7701细胞,P<0.01),及诱导细胞周期阻滞在G1期(P<0.01)。研究结果表明,miR-486-5p在肝癌组织中低表达;重建肝癌细胞内miR-486-5p的表达,可有效诱导肝癌细胞周期阻滞和凋亡,从而抑制肝癌细胞生长,提示miR-486-5p可能在肝癌的发生发展中起着重要作用。This study aims to investigate the expression of miR-486-5p in hepatocellular carcinoma （HCC） and to explore its role in the growth of HCC cells. Firstly, the expression of miR-486-Sp in 40 samples of HCC tissue and their matched controls as well as in a group of HCC cell lines （QGY-7701 ,QGY-7703,BEL-7404,SMMC-7721 ,Huh7 ,HepG2 and PCL/PRF/5） was tested by qRT-PCR. Secondly, synthetic miR-486-5p mimics or negative control（NC） were respectively transfected into the above mentioned cell lines by liposome method, and their effect on HCC proliferation was tested by CCK8 assay. Last, the role of miR-486-5p on HCC cells cell cycle and apoptosis was detected by FACS. The results showed that. 1, Consistent with the result of miRomes MPSS, the expression of miR-486-5p was downregulated in 40 paired HCC tissues and HCC cells compared to that in the match control by qRT-PCR （P 〈0.05）. 2. CCK8 assay revealed that miR-486-5p can effectively and steadily inhibit proliferation of all the HCC cells tested （P 〈0.05）. 3. FACS technology found that miR-486-5p overexpression could induce HCC cells apoptosis （QGY-7703 cell P 〈0. 001; QGY-7701 cell P d0.01） and blocked cell cycle progression of HCC cells at G1 phage （P 〈0.01）. In conclusion, these results indicate that miR-486-5p is markedly decreased in HCC,and its restoration can inhibit HCC cell proliferation by arresting cell cycle at Glphase and inducing apoptosis. These evidences suggest that miR-486-5p may play important roles in HCC development.

关 键 词：肝癌 miR-486—5p 细胞周期 凋亡 

分 类 号：R593[医药卫生—内科学]