Ou Chao, Peng Ning-Fu, Li Hang, Peng Yu-Chong, Li Le-Qun
Department of Clinical Laboratory Medicine, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning, China.
Department of Hepatobiliary Surgery, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning, China.
J Recept Signal Transduct Res. 2020 Apr;40(2):157-165. doi: 10.1080/10799893.2020.1721537. Epub 2020 Feb 4.
This study aimed to elucidate the regulatory role and molecular regulation mechanism of miR-130b gene in the process of invasion and metastasis of hepatocarcinoma, and provide a theoretical basis for seeking of effective prevention and treatment of new targets for hepatocellular carcinoma. The expression level of miR-130b gene in hepatocarcinoma tissues was determined by qRT-PCR. The biological function and mechanism of miR-130b gene were verified by cell and animal models, and the target gene was verified by double luciferase assay. In the liver cancer tissues of patients with metastasis, the expression level of miR-130b gene was increased, and the difference was significantly significant ( < 0.05). Evaluation of independent risk factors for overall survival showed significant difference ( < 0.01). Up-regulation of miR-130b in MHCC97L- subpopulation cells significantly enhanced the invasion and migration ability, and the difference was statistically significant ( < 0.05). The invasion and migration ability of MHCC97H + subpopulation cells with increased expression of miR-130b was significantly decreased, and the difference was notably significant ( < 0.05). When the expression of miR-130b in MHCC97H + subpopulation cells was inhibited, the expressions of Notch-Dll1 and SOX2, Nanog and E2F3 proteins in transplanted tumor tissues were significantly higher than those in other groups ( < 0.05). When miR-130b in MHCC97L- subpopulation cells was up-regulated, the expressions of Notch-Dll1 and Bcl-2, CCND1, Nanog and MET proteins in transplanted tumor tissues were significantly increased than those in other groups ( < 0.05). The prediction results of bioinformatics data suggest that the target gene of miR-130b may be Notch-Dll1 gene. The experiment of luciferase reporter gene confirmed that miR-130b gene can be inhibited and contains fluorescent reporter gene with complementary binding site, lost activity. The miR-130b gene can inhibit the protein expression of Notch-Dll1, and it can promote the invasion and metastasis of liver cancer cells.
本研究旨在阐明miR-130b基因在肝癌侵袭转移过程中的调控作用及分子调控机制，为寻找有效的肝癌防治新靶点提供理论依据。采用qRT-PCR检测肝癌组织中miR-130b基因的表达水平。通过细胞和动物模型验证miR-130b基因的生物学功能及机制，并采用双荧光素酶报告基因实验验证靶基因。在发生转移的肝癌患者组织中，miR-130b基因表达水平升高，差异具有统计学意义（<0.05）。评估总生存的独立危险因素显示差异有统计学意义（<0.01）。上调MHCC97L-亚群细胞中miR-130b的表达可显著增强侵袭和迁移能力，差异有统计学意义（<0.05）。miR-130b表达升高的MHCC97H+亚群细胞侵袭和迁移能力显著降低，差异有统计学意义（<0.05）。抑制MHCC97H+亚群细胞中miR-130b的表达后，移植瘤组织中Notch-Dll1、SOX2、Nanog和E2F3蛋白的表达显著高于其他组（<0.05）。上调MHCC97L-亚群细胞中miR-130b的表达后，移植瘤组织中Notch-Dll1、Bcl-2、CCND1、Nanog和MET蛋白的表达显著高于其他组（<0.05）。生物信息学数据预测结果提示，miR-130b的靶基因可能为Notch-Dll1基因。荧光素酶报告基因实验证实，miR-130b基因可抑制含有与荧光报告基因互补结合位点的荧光素酶活性。miR-130b基因可抑制Notch-Dll1蛋白表达，促进肝癌细胞侵袭转移。
