HNRNPA1通过下调SLC7A11-GPX4轴促进胃癌MKN74细胞铁死亡

HNRNPA1 promotes ferroptosis in human gastric cancer MKN74 cells via downregulation of the SLC7A11-GPX4 axis

摘要

目的 探讨异质性核糖核蛋白A1（HNRNPA1）在胃癌MKN74细胞铁死亡中的作用及机制。方法 通过生物信息学分析TCGA数据库HNRNPA1与铁死亡相关基因的表达相关性;采用比色法检测细胞内亚铁离子（Fe²⁺）水平;采用DCFH-DA荧光探针法检测细胞内活性氧（ROS）水平;CCK-8法测定细胞活力;Western blot法检测谷胱甘肽过氧化物酶4（GPX4）、溶质载体家族7成员11（SLC7A11）蛋白表达水平,并通过Ferrostatin-1（Fer-1）进行铁死亡抑制实验。结果 生信分析显示HNRNPA1在胃癌组织中高表达,并与SLC7A11呈正相关;敲低HNRNPA1可显著增加MKN74细胞Fe²⁺积累和ROS水平,降低细胞活力,同时下调SLC7A11与GPX4蛋白表达,Fer-1处理可部分逆转上述表型。结论 HNRNPA1敲低可通过下调SLC7A11-GPX4轴,促进细胞Fe²⁺积累和ROS水平升高,从而触发铁死亡。本研究揭示HNRNPA1在胃癌MKN74细胞铁死亡中的调控作用,为靶向HNRNPA1的新型治疗策略提供实验依据。

Abstract

Objective To investigate the role and underlying mechanism of heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1) in ferroptosis of human gastric cancer MKN74 cells. Methods Bioinformatic analysis was performed to assess the correlation between HNRNPA1 and ferroptosis-related genes in TCGA gastric cancer dataset. Intracellular Fe²⁺ levels were measured by colorimetric assay, and ROS levels were measured using DCFH-DA fluorescent probe and fluorescence microscopy.Cell viability was determined by CCK-8 assay, and protein expression of SLC7A11 and GPX4 was detected by Western blot. Ferrostatin-1 (Fer-1) was used to inhibit ferroptosis and assess rescue effects. Results Bioinformatic analysis indicated that HNRNPA1 was highly expressed in gastric cancer tissues and positively correlated with SLC7A11. Knockdown of HNRNPA1 significantly increased intracellular Fe²⁺ accumulation and ROS levels, reduced cell viability, and downregulated SLC7A11 and GPX4 expression, while Fer-1 treatment partially reversed these effects. Conclusion HNRNPA1 knockdown promotes ferroptosis in MKN74 cells by downregulating the SLC7A11-GPX4 axis, leading to Fe²⁺ accumulation and ROS elevation. This study elucidates the regulatory role of HNRNPA1 in gastric cancer MKN74 cells ferroptosis and provides experimental evidence for potential HNRNPA1-targeted therapeutic strategies.

关键词

HNRNPA1 / 胃癌 / 铁死亡 / SLC7A11 / GPX4

Key words

HNRNPA1 / gastric cancer / ferroptosis / SLC7A11 / GPX4

[1] Bray F, Laversanne M, Sung H, et al.Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2024, 74(3): 229-263.
[2] Chen Y, Jia K, Xie Y, et al.The current landscape of gastric cancer and gastroesophageal junction cancer diagnosis and treatment in China: a comprehensive nationwide cohort analysis[J]. J Hematol Oncol, 2025, 18(1): 42-56.
[3] Dixon SJ, Lemberg KM, Lamprecht MR, et al.Ferroptosis: an iron-dependent form of nonapoptotic cell death[J]. Cell, 2012, 149(5): 1060-1072.
[4] Li J, Cao F, Yin H liang, et al. Ferroptosis: past, present and future[J]. Cell Death Dis, 2020, 11(2): 88-100.
[5] Chen F, Kang R, Tang D, et al.Ferroptosis: principles and significance in health and disease[J]. J Hematol Oncol, 2024, 17(1): 41-81.
[6] Feng J, Zhou J, Lin Y, et al.hnRNP A1 in RNA metabolism regulation and as a potential therapeutic target[J]. Front Pharmacol, 2022, 13: 986409.
[7] Chen Y, Liu J, Wang W, et al.High expression of hnRNPA1 promotes cell invasion by inducing EMT in gastric cancer[J]. Oncol Rep, 2018: 1-9.
[8] Jiang C, Xu D, Feng H, et al.hnRNPA1 promotes the metastasis and proliferation of gastric cancer cells through WISP2-guided wnt/β-catenin signaling pathway[J]. Discov Oncol, 2024, 15(1): 465.
[9] Wang JM, Liu N, Wei XJ, et al.Regulation of AUF1 alternative splicing by hnRNPA1 and SRSF2 modulate the sensitivity of ovarian cancer cells to cisplatin[J]. Cell Oncol, 2024, 47(6): 2349-2366.
[10] Ding X, Cui L, Mi Y, et al.Ferroptosis in cancer: revealing the multifaceted functions of mitochondria[J]. Cell Mol Life Sci, 2025, 82(1): 277-297.
[11] Zhou Q, Meng Y, Li D, et al.Ferroptosis in cancer: from molecular mechanisms to therapeutic strategies[J]. Signal Transduct Target Ther, 2024, 9(1): 55-84.
[12] Zhu WW, Liu Y, Yu Z, et al.SLC7A11-mediated cell death mechanism in cancer: a comparative study of disulfidptosis and ferroptosis[J]. Front Cell Dev Biol, 2025, 13: 1559423.
[13] Chen H, Wang C, Liu Z, et al.Ferroptosis and its multifaceted role in cancer: mechanisms and therapeutic approach[J]. Antioxidants, 2022, 11(8): 1504.
[14] Cui K, Wang K, Huang Z.Ferroptosis and the tumor microenvironment[J]. J Exp Clin Cancer Res, 2024, 43(1): 315-331.
[15] Lee J, Roh JL.SLC7A11 as a gateway of metabolic perturbation and ferroptosis vulnerability in cancer[J]. Antioxidants, 2022, 11(12): 2444.
[16] 李瑞杰. 铁死亡的分子机制及其对膀胱癌的影响[J]. 中南大学学报(医学版),2024,49(2):286-295.
[17] Qiao Y, Su M, Zhao H, et al.Targeting FTO induces colorectal cancer ferroptotic cell death by decreasing SLC7A11/GPX4 expression[J]. J Exp Clin Cancer Res, 2024, 43(1): 108-125.