Zhang Jiang-Yuan, Nie Xue-Kun, Chen Zi-Chun, Zhou Su-Juan, Lin Xiao-Hui, Zhang Li, Zhong Di, Xiao Bing-Ying, Jiang Shi-Qing, Huang Wei-Ying, Lin Min-Hua, Wang Yu-Jia
Ningde Municipal Hospital of Ningde Normal University, Ningde, Fujian, China.
Shanghai General Hospital Ningde Hospital, Ningde, Fujian, China.
Exp Dermatol. 2025 Aug;34(8):e70145. doi: 10.1111/exd.70145.
Previous studies have unequivocally established the efficacy of gefitinib, an Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor (EGFR-TKIs), in the management of patients afflicted with advanced Non-Small Cell Lung Cancer (NSCLC). Nonetheless, the manifestation of cutaneous toxicities of varying severity has been observed to compromise patient survival outcomes and limit its clinical applicability. Elucidating the mechanistic underpinnings of gefitinib-induced dermal barrier dysfunction is imperative, as it holds the potential to inform future therapeutic strategies and facilitate the development of innovative pharmacological interventions. Utilising a multi-modal approach, this study employed network pharmacology and molecular docking techniques to identify potential etiological factors of gefitinib-induced dermal barrier dysfunction. Oral administration of gefitinib was conducted to establish a clinical model, followed by Haematoxylin and Eosin (HE) staining for epidermal and stratum corneum morphological assessment, and immunohistochemical quantification of Keratins 1 (K1) and K10 and Desmoglein-1 (DSG-1). Molecular expression levels of K10, K17, Claudin-4 (CLDN4), Interleukin-6 (IL-6), Tumour Necrosis Factor-α (TNF-α), and Ephrin Type-A Receptor 2 (EPHA2) were evaluated in HaCaT keratinocytes using Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) and Western Blot assays. Additionally, EPHA2 mRNA and protein expression in murine cutaneous tissues were ascertained through RT-qPCR and Western Blot analyses. Network pharmacology and molecular docking implicated EPHA2 as a central mediator in gefitinib-induced epidermal barrier dysfunction pathways. In murine models, gefitinib administration resulted in palpebral desquamation and dorsal cutaneous erythema, accompanied by elevated expression of K1, K17, and DSG-1 and epidermal hyperplasia. Furthermore, gefitinib augmented K10, K17, IL-6, and TNF-α expression in HaCaT cells, significantly attenuated cellular viability, and suppressed CLDN4 expression. EPHA2 mRNA and protein expression were notably downregulated in both HaCaT cells and BALB/c murine models. Ephrin-A1 Fc, an EPHA2 agonist, effectively mitigated gefitinib-induced cutaneous damage and inflammation, while concurrently downregulating K10, K17, IL-6, and TNF-α expression in HaCaT cells and upregulating CLDN4 expression. Gefitinib appears to induce dermal barrier dysfunction via the downregulation of EPHA2 expression.
先前的研究已明确证实表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs）吉非替尼在晚期非小细胞肺癌（NSCLC）患者治疗中的疗效。然而，已观察到不同严重程度的皮肤毒性表现会影响患者的生存结果并限制其临床应用。阐明吉非替尼诱导皮肤屏障功能障碍的机制基础至关重要，因为这有可能为未来的治疗策略提供信息，并促进创新药物干预措施的开发。本研究采用多模态方法，运用网络药理学和分子对接技术来确定吉非替尼诱导皮肤屏障功能障碍的潜在病因。通过口服吉非替尼建立临床模型，随后进行苏木精和伊红（HE）染色以评估表皮和角质层形态，并对角蛋白1（K1）、K10和桥粒芯糖蛋白-1（DSG-1）进行免疫组织化学定量分析。使用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法评估HaCaT角质形成细胞中K10、K17、紧密连接蛋白-4（CLDN4）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和Ephrin A型受体2（EPHA2）的分子表达水平。此外，通过RT-qPCR和蛋白质免疫印迹分析确定小鼠皮肤组织中EPHA2的mRNA和蛋白表达。网络药理学和分子对接表明EPHA2是吉非替尼诱导的表皮屏障功能障碍途径中的核心介质。在小鼠模型中，给予吉非替尼会导致睑部脱屑和背部皮肤红斑，同时伴有K1、K17和DSG-1表达升高以及表皮增生。此外，吉非替尼增加了HaCaT细胞中K10、K17、IL-6和TNF-α的表达，显著降低细胞活力，并抑制CLDN4表达。在HaCaT细胞和BALB/c小鼠模型中，EPHA2的mRNA和蛋白表达均明显下调。EPHA2激动剂Ephrin-A1 Fc有效减轻了吉非替尼诱导的皮肤损伤和炎症，同时下调了HaCaT细胞中K10、K17、IL-6和TNF-α的表达，并上调了CLDN4表达。吉非替尼似乎通过下调EPHA2表达诱导皮肤屏障功能障碍。
