miR-149-5p在肝细胞癌中的表达及其临床意义  

Expression of miR-149-5p in hepatocellular carcinoma and its clinical significance

作　　者：曹亚玲[1] 陈陵[1] 吕沐瀚[1] 魏晓林[2] 吴玉云[1] 罗刚[1] 刘祥德[2] 杨仕明[1] 

机构地区：[1]第三军医大学西南医院全军消化病研究所,重庆400038 [2]第三军医大学西南医院全军肝胆外科研究所,重庆400038

出　　处：《第三军医大学学报》2012年第7期627-631,共5页Journal of Third Military Medical University

摘　　要：目的分析miR-149-5p在肝细胞癌(hepatocellular carcinoma,HCC)组织中的表达及其临床意义。方法应用qRT-PCR方法检测40对HCC癌组织和癌旁肝组织中miR-149-5p的表达,并分析miR-149-5p的表达与肝癌患者临床病理之间的关系,进一步将以红色荧光蛋白(red fluorescent protein,RFP)为报告基因的miR-149-5p慢病毒(LV-miR-149-5p)感染MHCC97-H肝癌细胞,以过表达miR-149-5p,采用MTT检测感染后细胞增殖的变化,采用流式细胞仪测定感染后细胞周期的变化,并采用Transwell侵袭实验检测感染后细胞的体外侵袭能力的变化。结果 qRT-PCR结果提示miR-149-5p在HCC癌组织中的相对含量(5.522±1.104)低于癌旁肝组织(9.279±1.594)(P<0.05),统计分析表明miR-149-5p在癌和癌旁肝组织中相对含量的比值与AFP含量负相关(P<0.05)。体外实验证实,miR-149-5p过表达不能影响细胞周期但能抑制细胞增殖(P<0.05),而在Transwell侵袭实验中,LV-miR-149-5p感染MHCC97-H细胞后,对照细胞组透膜细胞数为(86.3±11.8)/视野,对照慢病毒组透膜细胞数为(83.7±4.0)/视野,LV-miR-149-5p组透膜细胞数为(50.6±3.4)/视野,实验组细胞侵袭能力较对照组细胞显著降低(P<0.05)。结论 miR-149-5p的过表达可抑制MHCC97-H细胞的增殖和侵袭,因此,miR-149-5p可作为一种抑癌基因在HCC的诊断及治疗中发挥作用。Objective To determine the expression of miR-149-5p in hepatocellular carcinoma（HCC） and its clinical significance.Methods The expression of miR-149-5p in 40 pairs of HCC tissue samples and adjacent noncancerous liver tissues were tested by quantitative real-time polymerase chain reaction（qRT-PCR）.The relationship between the expression and clinical pathological data was analyzed.MHCC97-H cells were infected by lentiviral vectors with red fluorescent proteins（RFP） for the reporter gene to over-express miR-149-5p（LV-miR-149-5p）.Then MTT assay was carried out to analyze cell proliferation.Flow cytometry was performed to analyze cell cycle.Transwell invasion assay was accomplished to analyze cell invasion ability.Results The results of qRT-PCR suggested that the relative expression of miR-149-5p in HCC（5.522±1.104） was lower than in the adjacent noncancerous liver tissues（9.279±1.594,P0.05）.Statistical analysis showed that the relative expression ratio of cancer and adjacent noncancerous liver tissues was correlated negatively with α-fetoprotein（AFP） levels（P0.05）.In vitro experiments confirmed that over-expression of miR-149-5p didn＇t affect the cell cycle but inhibited cell proliferation（P0.05）.And in transwell invasion assay,the cells through the matrigel were 86.3±11.8,83.7±4.0,50.6±3.4 in control group,control lentiviral treatment group,and LV-miR-149-5p treatment group,respectively.The invasive ability of experimental group was significantly reduced than that of control group（P0.05）.Conclusion Over-expression of miR-149-5p could inhibit the proliferation and invasion of MHCC97-H cells.Therefore,miR-149-5p might act as a tumor suppressor gene in HCC diagnosis and treatment.

关 键 词：miR-149-5p miR-149 肝癌 肝细胞癌 

分 类 号：R394.2[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]