Cancer Center, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Department of Hepatobiliary Surgery, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong, China.
J Cell Mol Med. 2019 Apr;23(4):2475-2488. doi: 10.1111/jcmm.14121. Epub 2019 Feb 1.
The study was aimed to screen out miRNAs with differential expression in hepatocellular carcinoma (HCC), and to explore the influence of the expressions of these miRNAs and their target gene on HCC cell proliferation, invasion and apoptosis. MiRNAs with differential expression in HCC were screened out by microarray analysis. The common target gene of these miRNAs (miR-139-5p, miR-940 and miR-193a-5p) was screened out by analysing the target genes profile (acquired from Targetscan) of the three miRNAs. Expression levels of miRNAs and SPOCK1 were determined by quantitative real time polymerase chain reaction (qRT-PCR). The target relationships were verified by dual luciferase reporter gene assay and RNA pull-down assay. Through 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,thiazolyl blue tetrazolium bromide (MTT) and transwell assays and flow cytometry, HCC cell viability, invasion and apoptosis were determined. In vivo experiment was conducted in nude mice to investigate the influence of three miRNAs on tumour growth. Down-regulation of miR-139-5p, miR-940 and miR-193a-5p was found in HCC. Overexpression of these miRNAs suppressed HCC cell viability and invasion, promoted apoptosis and inhibited tumour growth. SPOCK1, the common target gene of miR-139-5p, miR-940 and miR-193a-5p, was overexpressed in HCC. SPOCK1 overexpression promoted proliferation and invasion, and restrained apoptosis of HCC cells. MiR-139-5p, miR-940 and miR-193a-5p inhibited HCC development through targeting SPOCK1.
本研究旨在筛选出肝癌（HCC）中差异表达的 microRNAs，并探讨这些 microRNAs 的表达及其靶基因对 HCC 细胞增殖、侵袭和凋亡的影响。通过 microarray 分析筛选出 HCC 中差异表达的 microRNAs。通过分析这 3 个 microRNAs 的靶基因谱（从 Targetscan 获得）筛选出这 3 个 microRNAs 的共同靶基因（miR-139-5p、miR-940 和 miR-193a-5p）。通过定量实时聚合酶链反应（qRT-PCR）测定 microRNAs 和 SPOCK1 的表达水平。通过双荧光素酶报告基因检测和 RNA 下拉实验验证靶关系。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）和噻唑蓝溴化四唑（MTT）和 Transwell 检测以及流式细胞术测定 HCC 细胞活力、侵袭和凋亡。在裸鼠体内实验中研究了这 3 个 microRNAs 对肿瘤生长的影响。发现 HCC 中存在 miR-139-5p、miR-940 和 miR-193a-5p 的下调。这些 microRNAs 的过表达抑制 HCC 细胞活力和侵袭，促进凋亡并抑制肿瘤生长。SPOCK1，miR-139-5p、miR-940 和 miR-193a-5p 的共同靶基因，在 HCC 中过表达。SPOCK1 的过表达促进了 HCC 细胞的增殖和侵袭，同时抑制了 HCC 细胞的凋亡。miR-139-5p、miR-940 和 miR-193a-5p 通过靶向 SPOCK1 抑制 HCC 的发展。
