Li Zhuokai, Wu Lvzhong, Tan Wei, Zhang Kun, Lin Qiaomei, Zhu Jinde, Tu Chaoyong, Lv Xinliang, Jiang Chuan
Department of Hepatobiliary and Pancreatic Surgery, Lishui Municipal Central Hospital, Lishui 323000, Zhejiang Province, China.
Department of General Surgery, Qingyuan County People's Hospital, Lishui, Zhejiang Province, China.
Ann Hepatol. 2021 Jul-Aug;23:100345. doi: 10.1016/j.aohep.2021.100345. Epub 2021 Mar 31.
This study aimed to explore the functional mechanism of the miRNA-20b-5p/cytoplasmic polyadenylation element binding protein 3 (miR-20b-5p/CPEB3) axis in hepatocellular carcinoma (HCC) so as to provide a new idea for targeted therapy of HCC.
Bioinformatics analysis was employed to obtain markedly differentially expressed miRNAs and mRNAs in The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset, so as to find target miRNA and its target mRNA. Real-time quantitative PCR was conducted to detect miR-20b-5p and CPEB3 mRNA expression. Western blot was performed to determine CPEB3 protein expression. Dual-luciferase reporter assay was carried out to verify the targeting relationship between miR-20b-5p and CPEB3. Cell counting kit-8 assay, wound healing assay, Transwell invasion assay and flow cytometry were conducted to evaluate the proliferation, migration, invasion and apoptosis of HCC cells.
Bioinformatics analysis suggested that miR-20b-5p and CPEB3 were markedly highly and lowly expressed, respectively, in HCC tissue in TCGA-LIHC dataset. Over-expressing miR-20b-5p facilitated the proliferation, migration and invasion, and suppressed the apoptosis of HCC cells. Dual-luciferase reporter assay validated that there was a targeting relationship between miR-20b-5p and CPEB3. The inhibitory effect of CPEB3 over-expression on HCC cell proliferation, migration and invasion was reversed by over-expressing miR-20b-5p.
The present study proved that miR-20b-5p promotes HCC cell proliferation, migration and invasion by inhibiting CPEB3 expression, which may provide a theoretical basis for the prognosis and treatment of HCC patients.
本研究旨在探讨微小RNA-20b-5p/细胞质聚腺苷酸化元件结合蛋白3（miR-20b-5p/CPEB3）轴在肝细胞癌（HCC）中的功能机制，从而为HCC的靶向治疗提供新思路。
采用生物信息学分析从癌症基因组图谱-肝细胞癌（TCGA-LIHC）数据集中获取显著差异表达的微小RNA（miRNA）和信使核糖核酸（mRNA），以寻找靶标miRNA及其靶标mRNA。进行实时定量聚合酶链反应（PCR）检测miR-20b-5p和CPEB3 mRNA表达。采用蛋白质免疫印迹法检测CPEB3蛋白表达。进行双荧光素酶报告基因检测以验证miR-20b-5p与CPEB3之间的靶向关系。采用细胞计数试剂盒-8检测、伤口愈合检测、Transwell侵袭检测和流式细胞术评估HCC细胞的增殖、迁移、侵袭和凋亡。
生物信息学分析表明，在TCGA-LIHC数据集中，miR-20b-5p和CPEB3在HCC组织中分别显著高表达和低表达。过表达miR-20b-5p促进HCC细胞的增殖、迁移和侵袭，并抑制其凋亡。双荧光素酶报告基因检测验证了miR-20b-5p与CPEB3之间存在靶向关系。过表达miR-20b-5p可逆转CPEB3过表达对HCC细胞增殖、迁移和侵袭的抑制作用。
本研究证明miR-20b-5p通过抑制CPEB3表达促进HCC细胞增殖、迁移和侵袭，这可能为HCC患者的预后和治疗提供理论依据。
