Gao Dongkai, Zhou Zumo, Huang Heqing
Department of Infectious Diseases, Zhuji People's Hospital of Zhejiang Province, Shaoxing, China.
Front Genet. 2019 Dec 13;10:1274. doi: 10.3389/fgene.2019.01274. eCollection 2019.
The micro-RNA miR-30b-3p has been reported to play a crucial role in several cancers. However, the biological function of miR-30b-3p in hepatocellular carcinoma (HCC) is still unknown. RT-qPCR was employed to determine the expression of miR-30b-3p in HCC tissues and cells. The MTT assay, colony formation assay, and cell migration and invasion assay were employed to evaluate the role of miR-30b-3p in HCC cells. A dual-luciferase reporter assay was employed to verify the target of miR-30b-3p. Western blotting was employed to determine the expression of key molecular signal transducers along TRIM27-PI3K/Akt axis. Expression of miR-30b-3p was markedly decreased in HCC tissues and cells and positively correlated with higher overall survival. Moreover, miR-30b-3p overexpression significantly repressed cell viability, proliferation, migration, and invasion of HCC cells . Notably, we demonstrated that miR-30b-3p directly bound to the 3'-untranslated region of tripartite motif containing 27 (TRIM27) mRNA by downregulating the expression of TRIM27, which was demonstrated to be negatively correlated with miR-30b-3p expression. TRIM27 was demonstrated to have an oncogenic role in HCC cells by enhancing cell viability, proliferation, migration, and invasion. Finally, the miR-30-3p-TRIM27-PI3K/Akt axis was shown to play a crucial role in HCC cells . Our results indicated that miR-30-3p might act as a new biomarker for the future diagnosis and treatment HCC.
据报道,微小RNA miR-30b-3p在多种癌症中发挥关键作用。然而,miR-30b-3p在肝细胞癌(HCC)中的生物学功能仍不清楚。采用RT-qPCR检测miR-30b-3p在HCC组织和细胞中的表达。采用MTT法、集落形成试验以及细胞迁移和侵袭试验评估miR-30b-3p在HCC细胞中的作用。采用双荧光素酶报告基因试验验证miR-30b-3p的靶标。采用蛋白质免疫印迹法检测沿TRIM27-PI3K/Akt轴的关键分子信号转导蛋白的表达。miR-30b-3p在HCC组织和细胞中的表达明显降低,且与较高的总生存率呈正相关。此外,miR-30b-3p过表达显著抑制HCC细胞的活力、增殖、迁移和侵袭。值得注意的是,我们证明miR-30b-3p通过下调含三联基序蛋白27(TRIM27)的表达直接与TRIM27 mRNA的3'-非翻译区结合,TRIM27的表达与miR-30b-3p的表达呈负相关。通过增强细胞活力、增殖、迁移和侵袭,TRIM27在HCC细胞中具有致癌作用。最后,miR-30-3p-TRIM27-PI3K/Akt轴在HCC细胞中发挥关键作用。我们的结果表明,miR-30-3p可能作为未来HCC诊断和治疗的新生物标志物。