基于生物信息学分析结合RT-PCR检测的肝细胞癌组织中miRNA表达变化及意义

Changes and significance of miRNA expression in hepatocellular carcinoma based on bioinformatics combined with RT-PCR

目的探讨基于生物信息学分析结合RT-PCR检测的肝细胞癌(HCC)组织中微小RNA(miRNA)表达变化及意义。方法检索癌症基因组图谱(TCGA)数据库、高通量基因表达数据库(GEO)中HCC相关数据集,检索时间截至2018年2月,用生物信息学分析肿瘤组织与正常组织之间miRNA的表达差异,在GEO芯片集的HCC组织中验证差异表达的miRNA。用RT-PCR检测76例份HCC及癌旁组织中hsa-miR-191-5p、hsa-miR-221-5p表达,并分析二者表达与患者临床病理特征的关系。结果在GSE12717筛选出43个差异表达miRNA(P<0.05,|Fold change|≥1),GSE10694筛选出50个差异表达miRNA(P<0.05,|Fold change|≥0.5),在TCGA中筛选出101个差异表达miRNA(P<0.05,|Fold change|≥1)。hsa-miR-221-5p、hsa-miR-191-5p在两个GEO数据集中呈现差异表达,但在TCGA中未检测出二者表达差异。HCC及其癌旁组织中hsa-miR-191-5p相对表达量分别为6.03±1.82、0.93±0.08,hsa-miR-221-5p相对表达量分别为6.03±1.82、0.93±0.08。HCC组织中hsa-miR-191-5p、hsa-miR-221-5p表达均高于癌旁组织(P均<0.05)。hsa-miR-191-5p表达与HCC的TNM分期、转移复发有关(P均<0.05),hsa-miR-221-5p表达与HCC的TNM分期、转移复发、包膜浸润有关(P均<0.05);二者表达与患者其他临床病理特征无关(P均>0.05)。结论基于生物信息学分析结合RT-PCR检测的HCC组织中hsa-miR-191、hsa-miR-221呈高表达,二者可能参与HCC的恶性进展。

Objective To investigate the changes and significance of miRNA expression in hepatocellular carcinoma(HCC)tissues based on bioinformatics combined with RT-PCR.Methods The HCC-related datasets in the Cancer Genome Atlas(TCGA)database and the High-throughput Gene Expression Database(GEO)were searched.The deadline for our search was February 2018.The difference in expression of miRNA between tumor tissue and normal tissues was analyzed in the GEO chip.Differentially expressed miRNAs were verified in sets of HCC tissues.Real-time quantitative RT-PCR was used to detect the expression of hsa-miR-191-5p and hsa-miR-221-5p in 76 cases of HCC and adjacent tissues,and the relationship between the expression of hsa-miR-191-5p and hsa-miR-221-5p was analyzed.Results Forty-three differentially expressed miRNAs were screened out in GSE12717(P<0.05,|Fold change|≥1),50 differentially expressed miRNAs were screened out by GSE10694(P<0.05,|Fold change|≥0.5),and 101 differentially expressed miRNAs were screened out in TCGA(P<0.05,|Fold change|≥1).The hsa-miR-221-5p and hsa-miR-191-5p were differentially expressed in the two GEO datasets,but no differences were detected in TCGA.The expression of hsa-miR-191-5p in the HCC and its adjacent tissues was 6.03±1.82 and 0.93±0.08,respectively,and the expression of hsa-miR-221-5p was 6.03±1.82 and 0.93±0.08,respectively.The expression levels of hsa-miR-191-5p and hsa-miR-221-5p in the HCC tissues were higher than those in the adjacent tissues(both P<0.05).The hsa-miR-191-5p was related to TNM stage and metastasis and recurrence of HCC(all P<0.05).The expression of hsa-miR-221-5p was related to TNM stage,metastasis recurrence and capsule infiltration of HCC(all P<0.05),but their expression was not related to the other clinicopathological features(all P>0.05).Conclusion The hsa-miR-191 and hsa-miR-221 are highly expressed in HCC tissues based on bioinformatics analysis combined with RT-PCR,and they may participate in the malignant progression of HCC.