MicroRNA-34a调控c-Met表达对肝癌细胞的增殖及迁移和侵袭的影响  

MiR-34a inhibits cell invasion and proliferation of hepatocellular carcinoma by repressing c-Met signaling

作　　者：蒋伟 李涛[1] 王静静[1] 施晓 季国忠[1] JIANG Wei;LI Tao;WANG Jing-jing;SHI Xiao;JI Guo-zhong(Digestive Medical Center,Second Affiliated Hospital of Nanjing Medical University/Jiangsu Key Lab of Cancer Biomarkers,Prevention and Treatment,Collaborative Innovation Center for Cancer Personalized Medicine,Nanjing Medical University,Nanjing 210011,Jiangsu,China)

机构地区：[1]南京医科大学第二附属医院消化医学中心江苏省肿瘤个体化医学协同创新中心,南京210011

出　　处：《医学研究生学报》2020年第2期139-143,共5页Journal of Medical Postgraduates

基　　金：江苏省科学技术厅重点研发专项基金(BE2016799)。

摘　　要：目的 MicroRNA-34a在原发性肝癌中的作用及其相关机制仍有待探究。文中旨在探讨miR-34a对肝癌增殖、迁移和侵袭能力的影响。方法使用脂质体转染试剂2000瞬时转染miR34a过表达、过表达对照质粒,建立miR-34a过表达的肝癌细胞HepG2和Huh-7细胞系,分为过表达组(转染过表达质粒miR-34a mimics)、过表达对照组(转染过表达对照质粒miR-34a mimics control)和空白组(未转染质粒)。qPCR检测各组细胞miR-34a的表达水平;使用CCK8增殖实验、克隆形成实验和Transwell实验,体外水平检测各组细胞的增殖、迁移和侵袭能力的变化;构建裸鼠肝癌皮下移植瘤模型,体内水平检测miR-34a对肝癌移植瘤的治疗作用。另将7 d后成瘤裸鼠随机分为miR-34a组(注射miR-34a micmics)和对照组(注射miR-34a micmics control),每组6只。荧光素酶报告基因实验鉴定miR-34a在肝癌中的靶基因。结果过表达组HepG2、Huh-7细胞miR-34a表达水平显著高于过表达对照组、空白组(P<0.01)。过表达组48、72、96 h细胞增殖活力较过表达对照组显著降低(P<0.01)。过表达组HepG2、Huh-7细胞的克隆形成数[(28.00±4.36)个、(51.33±9.56)个]较过表达对照组[(52±5.81)个、(97.33±6.74)个]显著降低(P<0.05)。过表达组HepG2、Huh-7细胞迁移能力、侵袭能力较过表达对照组显著降低(P<0.01)。成瘤14 d后,过表达组移植瘤重量显著低于过表达对照组的[(45.488±11.651)mg vs(125.408±27.107) mg,P<0.01],肿瘤体积亦显著低于过表达对照组(P<0.01)。miR-34a组pGL3-c-Met的荧光素酶活性较对照组显著升高[(0.717±0.052)vs(0.517±0.098),P<0.05]。结论肝癌中miR-34a水平上调可以抑制肝癌细胞的增殖、迁移和侵袭能力,其对肝癌细胞侵袭性生长的调控作用可能通过靶向下调c-Met表达实现。Objective To explore the role of miR-34 a in the proliferation, migration and invasion of hepatocellular carcinoma(HCC) cells. Methods HepG2 and Huh-7 cells were transfected with miR-34 a constructs by using lipofection. qPCR was applied to detect the expression of miR-34 a. Cell proliferation assay, colony formation assay, cell migration and invasion assay were used to elucidate the invasive growth of transfected and un-transfected cells. The subcutaneous xenograft in nude mice was used to verified the anti-tumor effect of miR-34 a in vivo. TargetScan, miR-WALK and luciferase assay were performed to explore the target of miR-34 a in HCC. Results Compared with the overexpression control group and blank group, the expression of miR-34 a in the miR-34 a overexpression group cells were(2.727±0.173) and(4.042±0.104), respectively(P<0.05). The levels of miR-34 a in overexpression control and blank groups were(0.003±0.030) and(0.005±0.027)(P>0.05). Compared with the overexpression control groups, cell proliferation, colony formation, migration and invasion of HCC cells overexpressing miR34 a were all impeded with statistically significant differences(P<0.05). In vivo, tumor weight and tumor volume were significantly reduced in the miR-34 a treatment group. The results of luciferase reporter gene indicated that c-Met was the direct target of miR-34 a in HCC. Conclusions Up-regulation of miR-34 a in hepatocellular carcinoma can inhibit the proliferation, migration and invasion of HCC cells, and its repression may be benefited by declining c-Met signaling in HCC.

关 键 词：肝癌 MIR-34A C-MET 侵袭性生长 

分 类 号：R735.7[医药卫生—肿瘤]