目的:本研究旨在探讨let-7c-5p对肝细胞癌(HCC)恶性行为的影响及其特异性分子通路。
方法:从癌症基因组图谱数据库获得let-7c-5p的差异表达和生存分析,然后通过qPCR初步验证其表达水平。let-7c-5p对HCC细胞恶性表型的影响随后使用CCK-8,transwell,伤口愈合,和流式细胞术测定。由let-7c-5p调节的下游mRNA被ENCORI数据库鉴定和确认,双荧光素酶报告基因,和蛋白质印迹分析。通过仙桃工具评估基因的免疫相关性,以及TIMER和TISIDB数据库。
结果:肝癌组织中let-7c-5p的表达水平明显降低,这被发现与肝癌患者的生存时间短密切相关。细胞表型实验表明let-7c-5p抑制增殖,入侵,和迁移,促进肝癌细胞凋亡。双荧光素酶报告基因和蛋白质印迹分析表明,CDCA8是let-7c-5p的下游mRNA,并受其负调控。挽救实验表明,CDCA8逆转了let-7c-5p对HCC细胞恶性表型的影响。此外,对公共数据库的分析表明,CDCA8与一些免疫细胞和免疫调节剂有关,它可能参与某些免疫途径和免疫功能的调节。
结论:Let-7c-5p已被证明通过下调免疫相关的CDCA8来抑制HCC,这将有助于了解HCC的发病机制并开发其治疗药物。
OBJECTIVE: The aim of this study is to investigate the effect of let-7c-5p on the malignant behaviors of hepatocellular carcinoma (HCC) and its specific molecular pathway.
METHODS: Differential expression and survival analysis of let-7c-5p were obtained from The Cancer Genome Atlas database, and then its expression level was preliminarily verified through qPCR. The effect of let-7c-5p on the malignant phenotype of HCC cells was subsequently evaluated using CCK-8, transwell, wound healing, and flow cytometry assays. Downstream mRNA regulated by let-7c-5p was identified and confirmed by ENCORI database, dual-luciferase reporter, and western blot assays. The immunocorrelation of genes was evaluated by Xiantao tool, and TIMER and TISIDB databases.
RESULTS: The expression level of let-7c-5p in HCC was obviously reduced, which was found to be closely associated with the short survival time of HCC patients. Cell phenotypic experiments showed that let-7c-5p inhibited proliferation, invasion, and migration and promoted apoptosis of HCC cells. Dual-luciferase reporter and western blot analysis demonstrated that CDCA8 is a downstream mRNA of let-7c-5p and is negatively regulated by it. Rescue experiment revealed that CDCA8 reversed the effect of let-7c-5p on the malignant phenotype of HCC cells. Furthermore, analysis of the public database revealed that CDCA8 is related to some immune cells and immunomodulators, and that it may participate in the regulation of some immune pathways and immune functions.
CONCLUSIONS: Let-7c-5p has been proved to suppress HCC by down-regulating immune-related CDCA8, which will help understand the pathogenesis of HCC and develop drugs for its treatment.