Uncovering the mechanism of ARRDC3 in non-small cell lung cancer cisplatin resistance and autophagy based on bioinformatics and experimental verification

IF 1.6 4区医学 Q4 CHEMISTRY, MEDICINAL

Letters in Drug Design & Discovery Pub Date : 2025-07-01 DOI:10.1016/j.lddd.2025.100097

Wenjun Tang, Jingru Luo, Shu Lin, Qixing Yan

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引用次数: 4

Abstract

Cisplatin (DDP) is a commonly used first-line chemotherapy drug for non-small cell lung cancer (NSCLC), but the emergence of DDP resistance greatly reduces its therapeutic effect. Arrestin domain-containing 3 (ARRDC3) has been found to play an anti-cancer role in NSCLC, but whether ARRDC3 regulates DDP resistance in NSCLC is still unclear. The levels of ARRDC3, myeloid ecotropic insertion site 1 (MEIS1), protein kinase AMP-activated catalytic subunit alpha 2 (PRKAA2), and autophagy-related proteins were detected by quantitative real-time PCR (qRT-PCR) or western blot. Cell counting kit 8 (CCK8) assay, 3-(4, 5-dimethyl-2-thiazolyl)−2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay, 5-ethynyl-2′-deoxyuridine (EdU) assay, flow cytometry, and transwell assay were used to measure cell DDP resistance, proliferation, apoptosis and invasion. The interaction between ARRDC3 and MEIS1 or PRKAA2 was confirmed by dual-luciferase reporter assay, chromatin immunoprecipitation (ChIP) assay and Co-immunoprecipitation (Co-IP) assay. Xenograft tumor models were constructed to test the effect of ARRDC3 on NSCLC tumorigenesis and DDP resistance in vivo. ARRDC3 expression was downregulated in DDP-resistant NSCLC tissues and cells. Functional experiments showed that ARRDC3 overexpression restrained DDP resistance, growth, invasion and autophagy in NSCLC cells. Moreover, MEIS1 promoted ARRDC3 transcription by binding to its promoter region, and the inhibitory effects of MEIS1 overexpression on cell DDP resistance, growth, invasion and autophagy were reversed by ARRDC3 knockdown. Besides, ARRDC3 could interact with PRKAA2, and PRKAA2 overexpression also eliminated the regulation of ARRDC3 on cell DDP resistance, growth, invasion and autophagy. Furthermore, MEIS1 decreased PRKAA2 expression by activating ARRDC3 transcription. Additionally, upregulation of ARRDC3 reduced NSCLC tumorigenesis and DDP resistance in vivo. MEIS1-activated ARRDC3 represses NSCLC cell DDP resistance, growth, invasion and autophagy by reducing PRKAA2 expression, confirming the crucial role of MEIS1/ARRDC3/PRKAA2 axis in the chemotherapy resistance of NSCLC.

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基于生物信息学和实验验证，揭示ARRDC3在非小细胞肺癌顺铂耐药和自噬中的作用机制

顺铂（DDP）是非小细胞肺癌（NSCLC）常用的一线化疗药物，但DDP耐药的出现大大降低了其治疗效果。含有阻滞蛋白结构域3 （ARRDC3）在非小细胞肺癌中具有抗癌作用，但ARRDC3是否调控非小细胞肺癌的DDP耐药尚不清楚。采用实时荧光定量PCR （qRT-PCR）或western blot检测各组小鼠ARRDC3、髓系亲生态插入位点1 （MEIS1）、蛋白激酶amp活化的催化亚基α 2 （PRKAA2）和自噬相关蛋白的表达水平。采用细胞计数试剂盒8 （CCK8）法、3-（4,5 -二甲基-2-噻唑基）−2,5 -二苯基-2-四溴唑（MTT）法、5-乙基-2 ' -脱氧尿苷（EdU）法、流式细胞术和transwell法检测细胞对DDP的抗性、增殖、凋亡和侵袭。通过双荧光素酶报告试验、染色质免疫沉淀（ChIP）试验和共免疫沉淀（Co-IP）试验证实了ARRDC3与MEIS1或PRKAA2之间的相互作用。构建异种移植肿瘤模型，在体内检测ARRDC3对NSCLC肿瘤发生和DDP耐药的影响。在耐ddp NSCLC组织和细胞中，ARRDC3表达下调。功能实验表明，ARRDC3过表达抑制了NSCLC细胞对DDP的抵抗、生长、侵袭和自噬。此外，MEIS1通过结合ARRDC3的启动子区促进了ARRDC3的转录，并且MEIS1过表达对细胞DDP抗性、生长、侵袭和自噬的抑制作用被ARRDC3敲低逆转。此外，ARRDC3可以与PRKAA2相互作用，PRKAA2过表达也消除了ARRDC3对细胞DDP抗性、生长、侵袭和自噬的调节。此外，MEIS1通过激活ARRDC3转录来降低PRKAA2的表达。此外，ARRDC3的上调减少了体内NSCLC的肿瘤发生和DDP耐药性。MEIS1激活的ARRDC3通过降低PRKAA2表达抑制NSCLC细胞DDP耐药、生长、侵袭和自噬，证实了MEIS1/ARRDC3/PRKAA2轴在NSCLC化疗耐药中的关键作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Letters in Drug Design & Discovery 医学-医药化学

CiteScore

1.80

自引率

10.00%

发文量

245

审稿时长

3 months

期刊介绍： Aims & Scope Letters in Drug Design & Discovery publishes letters, mini-reviews, highlights and guest edited thematic issues in all areas of rational drug design and discovery including medicinal chemistry, in-silico drug design, combinatorial chemistry, high-throughput screening, drug targets, and structure-activity relationships. The emphasis is on publishing quality papers very rapidly by taking full advantage of latest Internet technology for both submission and review of manuscripts. The online journal is an essential reading to all pharmaceutical scientists involved in research in drug design and discovery.