南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (1): 17-24.doi: 10.12122/j.issn.1673-4254.2024.01.03

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过表达LncRNA MEG3通过促进铁死亡增强肝癌细胞对顺铂的化疗敏感性

朱 权，黄柏胜，位磊艳，罗奇志   

  1. 中南大学基础医学院免疫学系，生理学系，湖南 长沙 410008
  • 发布日期:2024-01-24

Overexpression of LncRNA MEG3 promotes ferroptosis and enhances chemotherapy sensitivity of hepatocellular carcinoma cells to cisplatin

ZHU Quan, HUANG Baisheng, WEI Leiyan, LUO Qizhi   

  1. Department of Immunology, Department of Physiology, School of Basic Medical Sciences, Central South University, Changsha 410008, China
  • Published:2024-01-24

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摘要： 目的 探讨过表达LncRNA MEG3对肝癌细胞HepG2和LM3增殖、迁移和顺铂化疗敏感性的影响及机制。方法 生物信息学在线网站分析MEG3在健康人群与肝细胞癌（HCC）患者的表达情况；将HepG2和LM3细胞各自分成2组，NC组：转染pcDNA3.1(+)空载体的细胞；MEG3-OE组：转染pcDNA3.1(+)-MEG3质粒的细胞。另外在检测活性氧（ROS）和丙二醛（MDA）实验中，将上述两组细胞分别各自给予顺铂（DDP）或铁死亡抑制剂Fer-1处理，实时荧光定量聚合酶链反应（qRT-PCR）检测细胞中MEG3的表达；CCK8和划痕实验检测肝癌细胞的增殖和迁移；另外采用CCK8实验检测DDP对肝癌细胞的抑制率；活性氧荧光探针（DCFH-DA）检测肝癌细胞中ROS的表达，MDA试剂盒检测肝癌细胞中MDA的浓度；Western blotting实验检测铁死亡相关蛋白谷胱甘肽过氧化物酶4（GPX4）和铁蛋白重链1（FTH1）的蛋白表达水平。结果 MEG3在肝癌细胞中的表达显著低于LO2细胞，与生物信息学分析结果一致（P<0.05）；与阴性对照组相比，MEG3-OE组肝癌细胞增殖迁移能力降低（P<0.05）、DDP的抑制率增高（P<0.05）、ROS水平升高、MDA表达水平升高（P<0.05）；MEG3-OE组肝癌细胞GPX4和FTH1蛋白表达均显著降低。结论 肝癌细胞中LncRNA-MEG3表达降低，过表达MEG3可抑制肝癌细胞增殖与迁移，增强DDP的化疗敏感性，其机制与促进肝癌细胞铁死亡有关。

关键词: 肝细胞癌；LncRNA MEG3；顺铂；化疗敏感性；铁死亡

Abstract: Objective To investigate the effect of overexpression of LncRNA MEG3 on proliferation, migration and cisplatin sensitivity of hepatoma cells HepG2 and LM3 and explore the underlying and mechanism. Methods The expression of MEG3 in healthy individuals and patients with hepatocellular carcinoma (HCC) was analyzed by online bioinformatics analysis, and Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect MEG3 expression in different HCC cell lines. A MEG3-overexpresing plasmid was transfected in HepG2 and LM3 cells, and the changes in cell proliferation and migration were examined using CCK8 assay and scratch assay. CCK8 assay was used to determine the inhibitory rate of cisplatin on the transfected cells. A reactive oxygen species (ROS) fluorescence probe (DCFH-DA) and malondialdehyde (MDA) kit were used to assess the changes in ROS production and MDA level in the cells. Western blotting was performed to detect the expression levels of ferroptosis-related proteins glutathione peroxidase 4 (GPX4) and ferritin heavy chain 1 (FTH1). Results The expression of MEG3 was significantly lower in HCC cells than in LO2 cells, which was consistent with the results of bioinformatic analysis (P<0.05). Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration (P<0.05), increased the cell inhibition rate of cisplatin (P<0.05), enhanced cellular ROS production and increased MDA levels in the cells (P<0.05). MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines. Conclusion The expression of MEG3 is decreased in HCC cells, and its overexpression inhibits proliferation and migration and enhances cisplatin sensitivity of HCC cells by promoting ferroptosis of the cells.

Key words: hepatocellular carcinoma; LncRNA MEG3; cisplatin; sensitivity to chemotherapy; ferroptosis