Zhang H, Wang Y, Ding H
Department of Gastroenterology, The Second Affiliated Hospital of Nanchang University, Nanchang, 330006, Jiangxi, China.
Department of Gastroenterology, The Second Clinical Medical College of Nanchang University, Nanchang, 330006, Jiangxi, China.
Clin Transl Oncol. 2021 Oct;23(10):2078-2089. doi: 10.1007/s12094-021-02611-y. Epub 2021 Apr 23.
Collagen type IV alpha 1 (COL4A1) exerts tumor-promoting functions in several tumors. However, its role in liver cancer remains not fully understood. Hence, this study aims to investigate the role of COL4A1 in regulating liver cancer cell behaviors and to validate its upstream regulatory mechanism.
Expression of xeroderma pigmentosum D (XPD) and COL4A1 was examined by qRT-PCR and western blot. Cell proliferation, migration, and invasion were evaluated. The protein levels of N-cadherin, vimentin, and E-cadherin were determined by western blot to evaluate epithelial-mesenchymal transition (EMT). The interaction between miR-29a-3p and COL4A1 was analyzed by luciferase reporter assay.
COL4A1 overexpression significantly promoted cell proliferation, migration, invasion, and EMT in Hep3B cells. In contrast, COL4A1 silencing yielded the opposite effects in HepG2 cells. Expression of COL4A1 was increased, whereas expression of XPD and miR-29a-3p was decreased in HCC tissues compared to controls. COL4A1 mRNA level was negatively correlated with expression of XPD and miR-29a-3p in HCC tissues. Furthermore, XPD silencing-mediated up-regulation of COL4A1 expression was attenuated by miR-29a-3p mimic. Moreover, miR-29a-3p mimic inhibited Hep3B cell proliferation, migration, and invasion by directly targeting COL4A1.
COL4A1 is negatively regulated by XPD-miR-29a-3p axis and promotes liver cancer progression in vitro.
IV型胶原α1（COL4A1）在多种肿瘤中发挥促肿瘤作用。然而，其在肝癌中的作用仍未完全明确。因此，本研究旨在探讨COL4A1在调控肝癌细胞行为中的作用，并验证其上游调控机制。
采用qRT-PCR和蛋白质免疫印迹法检测着色性干皮病D（XPD）和COL4A1的表达。评估细胞增殖、迁移和侵袭能力。通过蛋白质免疫印迹法检测N-钙黏蛋白、波形蛋白和E-钙黏蛋白的蛋白水平，以评估上皮-间质转化（EMT）。采用荧光素酶报告基因实验分析miR-29a-3p与COL4A1之间的相互作用。
COL4A1过表达显著促进Hep3B细胞的增殖、迁移、侵袭及EMT。相反，COL4A1沉默在HepG2细胞中产生相反的效果。与对照组相比，肝癌组织中COL4A1表达增加，而XPD和miR-29a-3p表达降低。肝癌组织中COL4A1 mRNA水平与XPD和miR-29a-3p的表达呈负相关。此外，miR-29a-3p模拟物减弱了XPD沉默介导的COL4A1表达上调。而且，miR-29a-3p模拟物通过直接靶向COL4A1抑制Hep3B细胞的增殖、迁移和侵袭。
COL4A1受XPD-miR-29a-3p轴负调控，并在体外促进肝癌进展。
