Sun Qi, Yang Xiansong, Wang Ye, Yang Kejia, Weng Yuan
Department of Thoracic Surgery, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China.
Department of Radiotherapy, Qingdao Central Hospital Affiliated to Qingdao University, Qingdao, Shandong, China.
Thorac Cancer. 2025 May;16(9):e70079. doi: 10.1111/1759-7714.70079.
Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related deaths worldwide, and cisplatin (DDP) resistance remains a significant challenge in NSCLC treatment.
Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to analyze NAT10 and tripartite motif containing 44 (TRIM44) mRNA levels. Western blotting assay was used to detect protein expression. Cell viability was analyzed by a cell counting kit-8 assay. Cell proliferation, apoptosis, invasion, and stem-like traits were assessed using a 5-Ethynyl-2'-deoxyuridineassay, flow cytometry, Transwell invasion assay, and sphere formation assay, respectively. The association between NAT10 and TRIM44 was identified by an RNA immunoprecipitation assay. A xenograft mouse model was established to evaluate the effect of NAT10 silencing on DDP sensitivity in vivo.
NAT10 expression was upregulated in DDP-resistant NSCLC tissues and cells. NAT10 knockdown enhanced DDP sensitivity in DDP-resistant NSCLC cells, accompanied by decreased protein expression of multidrug resistance 1 (MDR1). The silencing of NAT10 also inhibited the proliferation, invasion, and stem-like traits of DDP-resistant NSCLC cells, while inducing cell apoptosis. However, NAT10 overexpression displayed the opposite effects. Moreover, NAT10 maintained TRIM44 mRNA stability in an ac4C-dependent manner. TRIM44 overexpression reversed the NAT10 knockdown-induced effects on DDP sensitivity and the malignant progression of NSCLC cells. In addition, NAT10 silencing inactivated the PI3K/AKT pathway by regulating TRIM44 in DDP-resistant NSCLC cells. The treatment of the PI3K/AKT pathway inhibitor, LY294002, mitigated the effects of TRIM44 overexpression on DDP sensitivity and NSCLC cell progression. Further, NAT10 knockdown improved the sensitivity of tumors to DDP in vivo.
NAT10 knockdown improved DDP sensitivity in NSCLC by inhibiting the TRIM44/PI3K/AKT pathway, which may have significant clinical implications for overcoming DDP resistance in NSCLC treatment.
非小细胞肺癌（NSCLC）是全球癌症相关死亡的主要原因，顺铂（DDP）耐药仍是NSCLC治疗中的重大挑战。
采用定量逆转录聚合酶链反应（qRT-PCR）分析NAT10和含三联基序蛋白44（TRIM44）的mRNA水平。采用蛋白质免疫印迹法检测蛋白表达。用细胞计数试剂盒-8法分析细胞活力。分别采用5-乙炔基-2'-脱氧尿苷法、流式细胞术、Transwell侵袭试验和球体形成试验评估细胞增殖、凋亡、侵袭和干细胞样特性。通过RNA免疫沉淀试验确定NAT10与TRIM44之间的关联。建立异种移植小鼠模型以评估NAT10沉默对体内DDP敏感性的影响。
NAT10在DDP耐药的NSCLC组织和细胞中表达上调。NAT10敲低增强了DDP耐药NSCLC细胞对DDP的敏感性，同时多药耐药蛋白1（MDR1）的蛋白表达降低。NAT10沉默还抑制了DDP耐药NSCLC细胞的增殖、侵袭和干细胞样特性，同时诱导细胞凋亡。然而，NAT10过表达则表现出相反的效果。此外，NAT10以ac4C依赖的方式维持TRIM44 mRNA的稳定性。TRIM44过表达逆转了NAT10敲低对DDP敏感性和NSCLC细胞恶性进展的影响。此外，在DDP耐药NSCLC细胞中，NAT10沉默通过调节TRIM44使PI3K/AKT通路失活。PI3K/AKT通路抑制剂LY294002的处理减轻了TRIM44过表达对DDP敏感性和NSCLC细胞进展的影响。此外，NAT10敲低提高了体内肿瘤对DDP的敏感性。
NAT10敲低通过抑制TRIM44/PI3K/AKT通路提高了NSCLC对DDP的敏感性，这可能对克服NSCLC治疗中的DDP耐药具有重要的临床意义。
