miR-3934-5p靶向抑制β内酰胺酶表达促进肝癌细胞增殖

MiR-3934-5p promotes proliferation of hepatocellular carcinoma cells by targeting LACTB

目的观察miR-3934-5p在肝细胞癌(hepatocellular carcinoma, HCC)中的表达,探讨其临床意义及分子机制。方法收集53例2013年9月至2014年8月在本院确诊的肝细胞癌患者的癌旁组织及肿瘤组织。实时定量PCR(RT-qPCR)检测组织和细胞miR-3934-5p的表达;应用人类癌症基因库(the Cancer Genome Atlas, TCGA)分析正常肝组织与肝癌组织中miR-3934-5p的表达;用χ~2检验和Kaplan-Meier生存曲线分析miR-3934-5p与临床病理特征及生存预后之间的相关性;采用RT-qPCR和Western blot检测转染效率;CCK-8实验、平板克隆实验及EdU实验评价HCCLM3细胞的活性及增殖能力;分析HCC组织中β-内酰胺酶(beta-lactamases,LACTB) mRNA与miR-3934-5p表达的相关性;TargetScan数据库预测miR-3934-5p下游靶基因,通过荧光素酶报告基因实验加以验证。结果 miR-3934-5p在肝癌组织及细胞中高表达(P<0.05),且其高表达与肿瘤大小、肿瘤数量、血管侵犯及TNM分期显著相关(P<0.05);与miR-3934-5p低表达组相比,miR-3934-5p高表达组的肝癌患者5年生存率更低;CCK-8实验、平板克隆实验及EdU实验结果证实:敲减miR-3934-5p可显著抑制HCCLM3细胞的增殖能力(P<0.05);LACTB被证实是miR-3934-5p的直接作用靶点,且其介导了miR-3934-5p的生物学功能。结论 miR-3934-5p通过靶向抑制抑癌基因LACTB的表达促进肝癌细胞的增殖能力。

Objective To explore the expression,clinical significance and molecular mechanism of miR-3934-5p in hepatocellular carcinoma(HCC).Methods HCC tissues and adjacent normal tissues were collected from 53 HCC patients diagnosed in our hospital from September 2013 to August 2014.Real-time quantitative PCR(RT-qPCR)was performed to detect the level of miR-3934-5p in the above tissues and human liver cell line L02 and HCC cell lines.The Cancer Genome Atlas(TCGA)database was used to analyze the expression of miR-3934-5p in normal liver tissues and HCC tissues;Chi-square test with Kaplan-Meier survival curve were applied to analyze the correlation of miR-3934-5p and clinicopathological characteristics and survival prognosis;RT-qPCR and Western blotting were adopted to detect transfection efficiency;CCK-8 assay,colony formation assay and EdU assay were employed respectively to evaluate the activity and proliferation of HCCLM3 cells;Spearman correlation analysis was applied to study the correlation between LACTB mRNA and miR-3934-5p expression in HCC tissues;TargetScan database was used to predict the downstream target genes of miR-3934-5p,and the prediction results was verified by luciferase reporter assay.Results MiR-3934-5p was highly expressed in the liver cancer tissues and cells(P<0.05),and its high expression was significantly correlated with tumor size(P=0.040),tumor multiplicity(P=0.013),vascular invasion(P=0.006),and TNM stage(P=0.017);Compared to the low miR-3934-5p expression group,the HCC patients with high expression had poorer survival prognosis.CCK-8 assay,colony formation assay and EdU assay confirmed that knocking down miR-3934-5p notably inhibited the proliferation of HCCLM3 cells(P<0.05).LACTB had been proved to be the direct target of miR-3934-5p,and mediated the biological effect of miR-3934-5p on HCC cells.Conclusion MiR-3934-5p promotes the proliferation of HCC cells by targeting tumor suppressor gene LACTB.