摘要
目的探讨miR-542-5p靶向结合ENSA对肝细胞癌增殖、迁移和侵袭的影响。方法通过GSE36915数据矩阵分析miR-542-5p在肝癌组织和癌旁组织表达情况,构建miR-542-5p过表达肝癌细胞HepG2和Hep3B细胞系并使用RT-PCR验证,采用CCK-8、划痕、Transwell法检测细胞增殖、迁移和侵袭能力;使用miR-542-5p过表达Hep3B细胞系建立体内裸鼠移植瘤,观察移植瘤体积变化和重量变化,使用RT-PCR检测移植瘤miR-542-5p水平。通过GEO2R分析GSE101728、GSE7642的差异基因,使用miRWalk在线预测miR-542-5p的靶基因,筛选符合以上条件的基因得到ENSA;通过TCGA肝癌数据分析ENSA在肝癌组织和癌旁组织的表达情况,RT-PCR检测miR-542-5p过表达HepG2和Hep3B细胞ENSA水平,并在miR-542-5p过表达HepG2和Hep3B细胞系中转染ENSA过表达质粒,使用上述相同方法检测细胞增殖、迁移和侵袭能力。结果miR-542-5p在肝癌组织中表达水平显著低于癌旁组织(P<0.01),miR-542-5p过表达HepG2和Hep3B细胞系的光密度(A值)和相对细胞侵袭数均明显低于其阴性对照(miR-NC)(P<0.01),而划痕宽度明显高于其miR-NC(P<0.01);裸鼠移植瘤实验结果显示miR-542-5p过表达Hep3B细胞瘤的体积和重量明显低于miR-NC(P<0.01)。ENSA在肝癌组织中表达水平显著高于癌旁组织(P<0.01)且ENSA促进肝癌进展;miR-542-5p过表达HepG2和Hep3B细胞系中ENSA蛋白表达显著降低于其阴性对照(miR-NC)(P<0.01);miR-542-5p+ENSA双过表达HepG2和Hep3B细胞系的A值和细胞侵袭数均明显高于其miR-542-5p过表达细胞(P<0.01),而划痕宽度明显低于其miR-542-5p过表达细胞(P<0.01)。结论miR-542-5p可通过靶向抑制ENSA表达抑制肝细胞癌细胞增殖、迁移和侵袭。
Objective To explore the effect of miR-542-5p on proliferation,migration and invasion of hepatocellular carcinoma(HCC)by targeting ENSA.Methods The expression of miR-542-5p in HCC tissues and para-carcinoma tissues was analyzed by GSE36915 data matrix.miR-542-5p overexpression were performed in the HCC cell lines HepG2 and Hep3B and verified by RTPCR.The proliferation,migration and invasion of cells were detected by CCK-8,wound healing and Transwell assays.miR-542-5p overexpressed Hep3B cells were used to establish the transplanted tumor model in nude mice.The volume and weight of transplanted tumors were measured.The miR-542-5p expression level in transplanted tumors was detected by RT-PCR.The differential expression genes of GSE101728 and GSE7642 were analyzed by GEO2R,and the target genes of miR-542-5p were predicted by miRWalk,ENSA gene was then screened out.The expression level of ENSA in HCC tissues and para-carcinoma tissues was analyzed by TCGA HCC data.The expression level of ENSA in miR-542-5p overexpressed HepG2 and Hep3B cells was detected by RT-PCR.The cell proliferation,migration and invasion were detected in miR-542-5p overexpressed HepG2 and Hep3B cells after transfected with ENSA overexpression plasmid.Results The expression level of miR-542-5p in the HCC tissues was significantly lower than that in the para-carcinoma tissues(P<0.01).The proliferation rate,migration rate and relative number of invasion cells in the miR-542-5p overexpressed cells were significantly lower than those in miR-NC overexpressed cells(P<0.01).The results ofin vivoexperiments showed that the volume and weight of tumors in the miR-542-5p overexpression group were significantly lower than those in the miR-NC overexpression group(P<0.01).The expression level of ENSA in the HCC tissues was significantly higher than that in the para-carcinoma tissues(P<0.01),and ENSA could promote HCC progression.The protein expression level of ENSA in the miR-542-5p overexpressed cells were significantly lower than that in the miR-NC overexpr
作者
代汉莉
王国华
徐向勇
DAI Hanli;WANG Guohua;XU Xiangyong(Department of Clinical Laboratory,Wuhan Ninth Hospital,Wuhan,430081,China;Department of Oncology,the Second Staff Hospital of Wuhan Iron and Steel(Group)Company,Wuhan,430085,China;Department of Clinical Laboratory,the Second Staff Hospital of Wuhan Iron and Steel(Group)Company,Wuhan,430085,China)
出处
《医学分子生物学杂志》
CAS
2024年第5期405-412,共8页
Journal of Medical Molecular Biology
基金
湖北省自然科学基金(No.2017CFL083)。
