Hu Zongqiang, Yin Yanfeng, Jiang Jie, Yan Chuntao, Wang Yiting, Wang Dongdong, Li Li
Hepato-Pancreato-Biliary Surgery Department, The First People's Hospital of Kunming & The Calmette Affiliated Hospital of Kunming Medical University, Kunming, China.
The Central Laboratory, The First People's Hospital of Kunming & The Calmette Affiliated Hospital of Kunming Medical University, Kunming, China.
J Gastrointest Oncol. 2022 Apr;13(2):754-767. doi: 10.21037/jgo-21-916.
Most patients with hepatitis B virus (HBV) infection will develop hepatocellular carcinoma (HCC). This study aimed to explore the potential mechanism of miR-142-3p in HCC caused by HBV infection.
HepG2 cells and M1 macrophages were cocultured and then infected with HBV to establish an model. MicroRNA (miRNA) and messenger RNA (mRNA) expression was analyzed by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot. The protein expressions of COX2, ACSL4, PTGS2, GPX4, and NOX1 were analyzed by Western blot. Flow cytometry and TUNEL assays were used to assess cell reactive oxygen species (ROS) and ferroptosis, respectively. Cell invasion and migration were measured by Transwell assay. To evaluate the ferroptosis of M1-type macrophages, glutathione (GSH), malondialdehyde (MDA), and Fe content was detected by corresponding kits. Dual luciferase reporter gene detection verified the targeting relationship between miR-142-3p and SLC3A2.
MiR-142-3p was highly expressed in HBV-infected HCC patients and HBV-infected M1-type macrophages. Inhibition of miR-142-3p or overexpression of SLC3A2 reversed ferroptosis and inhibited the proliferation, migration, and invasion of HCC cells.
Our findings indicated that miR-142-3p promoted HBV-infected M1-type macrophage ferroptosis through SLC3A2, affecting the production of GSH, MDA, and Fe and accelerating the development of HCC. The regulation of miR-142-3p and its target genes will help to clarify the pathogenesis of HCC induced by HBV infection and provide new theoretical foundations and therapeutic targets.
大多数乙型肝炎病毒（HBV）感染患者会发展为肝细胞癌（HCC）。本研究旨在探讨miR-142-3p在HBV感染所致HCC中的潜在机制。
将HepG2细胞与M1巨噬细胞共培养，然后用HBV感染以建立模型。通过定量逆转录聚合酶链反应（RT-qPCR）和蛋白质印迹法分析微小RNA（miRNA）和信使核糖核酸（mRNA）表达。通过蛋白质印迹法分析COX2、ACSL4、PTGS2、GPX4和NOX1的蛋白表达。分别采用流式细胞术和TUNEL检测评估细胞活性氧（ROS）和铁死亡。通过Transwell检测法测量细胞侵袭和迁移。为评估M1型巨噬细胞的铁死亡，用相应试剂盒检测谷胱甘肽（GSH）、丙二醛（MDA）和铁含量。双荧光素酶报告基因检测验证miR-142-3p与SLC3A2之间的靶向关系。
miR-142-3p在HBV感染的HCC患者及HBV感染的M1型巨噬细胞中高表达。抑制miR-142-3p或过表达SLC3A2可逆转铁死亡，并抑制HCC细胞的增殖、迁移和侵袭。
我们的研究结果表明，miR-142-3p通过SLC3A2促进HBV感染的M1型巨噬细胞铁死亡，影响GSH、MDA和铁的产生，并加速HCC的发展。对miR-142-3p及其靶基因的调控将有助于阐明HBV感染所致HCC的发病机制，并提供新的理论基础和治疗靶点。
