Kang Xiaofeng, Huo Yan, Jia Songhao, He Fuliang, Li Huizi, Zhou Qing, Chang Nijia, Liu Donghui, Li Rongkuan, Hu Yi, Zhang Ping, Xu An
Department of Oncology, The Second Medical Center & National Clinical Research Center for Geriatric Diseases, Chinese PLA General Hospital, Beijing, China.
International School of Public Health and One Health, Hainan Medical University, Hainan, China.
Front Oncol. 2022 Jul 8;12:939605. doi: 10.3389/fonc.2022.939605. eCollection 2022.
Recently, long noncoding RNA LINC01134 has been shown to reduce cell viability and apoptosis the antioxidant stress pathway, thereby enhancing OXA resistance in hepatocellular carcinoma. However, the association of LINC01134 with ferroptosis and the underlying molecular mechanisms remain to be elucidated.
Bioinformatics analysis was employed to screen lncRNAs positively correlated with GPX4 and poor clinical prognosis. And Western blot and RT-PCR analysis in HCC cells confirmed the effect of LINC01134 on GPX4 expression. In addition, LINC01134 siRNA was transfected in HCC cells to detect the changes in cell viability, ROS, lipid peroxidation, MDA levels and GSH/GSSG levels. CCK-8, colony formation and apoptosis assays were performed to determine the effect of LINC01134 on cell death. The effect of LINC01134 and OXA on Nrf2 transcriptional binding to GPX4 was analyzed using dual luciferase reporter assay and CHIP. The expression of GPX4 and Nrf2 in HCC tissues was detected by FISH and IHC.
LINC01134 is a novel lncRNA positively correlated with GPx4 and associated with poor clinical prognosis. Silenced LINC01134 conferred OXA sensitivity by enhancing total ROS, lipid ROS, MDA levels and decreasing GSH/GSSG ratio. Mechanistically, LINC01134 and OXA could promote Nrf2 recruitment to the GPX4 promoter region to exert transcriptional regulation of GPX4. Clinically, LINC01134 was positively correlated with GPX4 or Nrf2, demonstrating the clinical significance of LINC01134, Nrf2 and GPX4 in OXA resistance of HCC.
We identified LINC01134/Nrf2/GPX4 as a novel and critical axis to regulate HCC growth and progression. Targeting GPX4, knocking down LINC01134 or Nrf2 could be a potential therapeutic strategy for HCC.
最近，长链非编码RNA LINC01134已被证明可通过抗氧化应激途径降低细胞活力并诱导凋亡，从而增强肝癌对奥沙利铂（OXA）的耐药性。然而，LINC01134与铁死亡的关联及其潜在分子机制仍有待阐明。
采用生物信息学分析筛选与谷胱甘肽过氧化物酶4（GPX4）呈正相关且临床预后不良的长链非编码RNA。肝癌细胞中的蛋白质免疫印迹法和逆转录-聚合酶链反应（RT-PCR）分析证实了LINC01134对GPX4表达的影响。此外，将LINC01134小干扰RNA（siRNA）转染至肝癌细胞中，以检测细胞活力、活性氧（ROS）、脂质过氧化、丙二醛（MDA）水平以及谷胱甘肽（GSH）/氧化型谷胱甘肽（GSSG）水平的变化。进行细胞计数试剂盒-8（CCK-8）、集落形成和凋亡检测，以确定LINC01134对细胞死亡的影响。使用双荧光素酶报告基因检测和染色质免疫沉淀（CHIP）分析LINC01134和奥沙利铂对核因子E2相关因子2（Nrf2）与GPX4转录结合的影响。通过荧光原位杂交（FISH）和免疫组织化学（IHC）检测肝癌组织中GPX4和Nrf2的表达。
LINC01134是一种与GPx4呈正相关且与临床预后不良相关的新型长链非编码RNA。沉默LINC01134可通过提高总ROS、脂质ROS、MDA水平并降低GSH/GSSG比值来赋予奥沙利铂敏感性。从机制上讲，LINC01134和奥沙利铂可促进Nrf2募集至GPX4启动子区域，从而对GPX4发挥转录调控作用。临床上，LINC01134与GPX4或Nrf2呈正相关，表明LINC01134、Nrf2和GPX4在肝癌奥沙利铂耐药中的临床意义。
我们确定LINC01134/Nrf2/GPX4是调节肝癌生长和进展的一个新的关键轴。靶向GPX4、敲低LINC01134或Nrf2可能是肝癌的一种潜在治疗策略。
