Zhu Q, Huang B, Wei L, Luo Q
Department of Immunology, School of Basic Medical Sciences, Central South University, Changsha 410008, China.
Department of Physiology, School of Basic Medical Sciences, Central South University, Changsha 410008, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Jan 20;44(1):17-24. doi: 10.12122/j.issn.1673-4254.2024.01.03.
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.
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).
The expression of MEG3 was significantly lower in HCC cells than in LO2 cells, which was consistent with the results of bioinformatic analysis ( < 0.05). Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration ( < 0.05), increased the cell inhibition rate of cisplatin ( < 0.05), enhanced cellular ROS production and increased MDA levels in the cells ( < 0.05). MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines.
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.
探讨长链非编码RNA MEG3过表达对肝癌细胞HepG2和LM3增殖、迁移及顺铂敏感性的影响，并探究其潜在机制。
通过在线生物信息学分析，分析健康个体和肝细胞癌（HCC）患者中MEG3的表达情况，并采用实时荧光定量PCR（qRT-PCR）检测不同HCC细胞系中MEG3的表达。将MEG3过表达质粒转染至HepG2和LM3细胞中，采用CCK8法和划痕实验检测细胞增殖和迁移的变化。采用CCK8法测定顺铂对转染细胞的抑制率。使用活性氧（ROS）荧光探针（DCFH-DA）和丙二醛（MDA）试剂盒评估细胞中ROS产生和MDA水平的变化。采用蛋白质免疫印迹法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）的表达水平。
HCC细胞中MEG3的表达明显低于LO2细胞，这与生物信息学分析结果一致（P<0.05）。在HCC细胞系中过表达MEG3可显著抑制细胞增殖和迁移（P<0.05），提高顺铂对细胞的抑制率（P<0.05），增强细胞ROS产生并增加细胞内MDA水平（P<0.05）。MEG3过表达显著降低了HCC细胞系中GPX4和FTH1的表达。
HCC细胞中MEG3表达降低，其过表达可通过促进细胞铁死亡抑制HCC细胞的增殖和迁移，并增强其对顺铂敏感性。
