内皮糖蛋白作为非小细胞肺癌培美曲塞敏感性的预测生物标志物：一项细胞研究

Endoglin as a predictive biomarker for pemetrexed sensitivity in non-small-cell lung cancer: a cellular study.

作者信息

Cheng Ching-Yuan, Chuang Wen-Chen, Lin Ching-Pin, Li Che-Hsing, Chang Hui-Yi, Wu Wen-Jun, Wu Ming-Fang, Ko Jiunn-Liang

机构信息

Division of Thoracic Surgery, Department of Surgery, Changhua Christian Hospital, Changhua, 500209, Taiwan.

Division of Thoracic Surgery, Department of Surgery, Yunlin Christian Hospital, Yunlin, 648106, Taiwan.

出版信息

Cancer Chemother Pharmacol. 2025 Jan 10;95(1):20. doi: 10.1007/s00280-024-04734-9.

DOI:10.1007/s00280-024-04734-9

PMID:39792181

Abstract

OBJECTIVE

Based on our previous research, which demonstrated that elevated plasma endoglin (ENG) levels in lung cancer patients were associated with a better prognosis, increased sensitivity to pemetrexed, and enhanced tumor suppression, this study aims to validate these findings at the cellular level. The focus is on membrane and extracellular ENG and their influence on drug response and tumor cell behavior in non-small cell lung cancer (NSCLC) cells.

METHODS

The correlation between ENG expression and pemetrexed-induced cytotoxicity in eight human non-squamous subtype NSCLC cell lines was analyzed. ENG in A549 and H1975 cells was knocked down using shRNA. MTT assay, cell cycle assay, western blot analysis, and boyden chamber assay were used to detect the effect of ENG on pemetrexed-induced cytotoxicity, cell cycle distribution, and cell migration.

RESULTS

The expression of membrane ENG was positively correlated with pemetrexed-induced cytotoxicity in human NSCLC cells. Compared to pemetrexed-sensitive A549 cells, the A549/a400 (pemetrexed-resistant subline) cells exhibited a reduced accumulation of cells in the S phase, making them less susceptible to cell death. ENG knockdown also alleviated pemetrexed-induced S phase arrest and regulated G1/S phase-related proteins (p53, p21, CDK2, and Cyclin A). Additionally, co-treatment with recombinant ENG enhanced pemetrexed-induced migration inhibition in the sensitive cel1 line and cytotoxicity in the resistance cell line.

CONCLUSION

The present results strengthened our prior clinical findings, showing that higher membrane ENG expression enhances pemetrexed-induced cytotoxicity and S phase arrest, which may involve the ENG-p21 pathway. Additionally, microenvironmental ENG enhanced the anti-migration of pemetrexed. These findings highlight the potential of ENG as a biomarker and therapeutic target, opening new avenues to improve the outcomes of non-squamous cell NSCLC treatment.

摘要

目的

基于我们之前的研究，该研究表明肺癌患者血浆中内皮糖蛋白（ENG）水平升高与较好的预后、对培美曲塞的敏感性增加以及增强的肿瘤抑制作用相关，本研究旨在在细胞水平上验证这些发现。重点是膜结合型和细胞外ENG及其对非小细胞肺癌（NSCLC）细胞药物反应和肿瘤细胞行为的影响。

方法

分析了8种人非鳞状亚型NSCLC细胞系中ENG表达与培美曲塞诱导的细胞毒性之间的相关性。使用shRNA敲低A549和H1975细胞中的ENG。采用MTT法、细胞周期分析法、蛋白质印迹分析和博伊登小室试验来检测ENG对培美曲塞诱导的细胞毒性、细胞周期分布和细胞迁移的影响。

结果

人NSCLC细胞中膜结合型ENG的表达与培美曲塞诱导的细胞毒性呈正相关。与对培美曲塞敏感的A549细胞相比，A549/a400（培美曲塞耐药亚系）细胞在S期的细胞积累减少，使其对细胞死亡的敏感性降低。ENG敲低也减轻了培美曲塞诱导的S期阻滞并调节了G1/S期相关蛋白（p53、p21、CDK2和细胞周期蛋白A）。此外，重组ENG联合治疗增强了培美曲塞对敏感细胞系的迁移抑制作用以及对耐药细胞系的细胞毒性。