Construction and mechanism of a small cell lung cancer cell line resistant to EP chemotherapy scheme
10.16016/j.2097-0927.202402065
- VernacularTitle:EP联合方案耐药小细胞肺癌细胞株的构建及机制探讨
- Author:
Mingpu LIU
1
,
2
;
Hongmei WANG
;
Yuanli WU
;
Duanfang ZHOU
;
Weiying ZHOU
Author Information
1. 400016 重庆,重庆医科大学 药学院药理学系
2. 400016 重庆,重庆医科大学 药物代谢研究重庆市重点实验室
- Keywords:
small cell lung cancer;
multidrug resistance;
etoposide;
cisplatin
- From:
Journal of Army Medical University
2024;46(18):2092-2100
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the etoposide (VP-16)combined with cisplatin (DDP) chemotherapy scheme (EP chemotherapy scheme)resistant small cell lung cancer (SCLC)cell line H446/EP and to identify the drug resistance characteristics and explore the mechanism.Methods NCI-H446 cells were treated with increasing concentrations of VP-16 and DDP to construct an H446/EP cell line.H446/EP and NCI-H446 cells were used as the research objects.The cell viability was detected by MTT assay,and the resistance index (RI)of H446/EP cells was calculated.Cell cloning assay and Incucyte cell proliferation (label-free)assay were used to detect cell proliferation ability.Transcriptome sequencing was performed to analyze the enrichment of differentially expressed genes (DEGs)in the 2 cell lines.Western blotting was applied to detect the protein expression levels of drug resistance,DNA damage repair (DDR),and autophagy markers.Results MTT assay showed that the resistance index (RI)of H446/EP cells to VP-16,DDP,and DOX were 6.14,3.43,and 1.96,respectively.The results of cell cloning assay and Incucyte cell proliferation assay indicated that the proliferation ability was significantly higher in the H446/EP cells than the NCI-H446 cells (P<0.01).Transcriptome sequencing and pathway enrichment analysis displayed that the DEGs between H446/EP and NCI-H446 cells were enriched in tumor chemoresistance,DDR,and autophagy pathways.Western blot results showed the expression levels of MRP1,BCRP,RAD51,γ-H2AX,and LC3-Ⅱ/LC3-Ⅰ were significantly increased,and that of p62 was obviously decreased in the H446/EP cells when compared with the NCI-H446 cells (P<0.05).Conclusion An EP chemotherapy-resistant H446/EP cell line is successfully constructed.Stronger proliferation ability,increased expression of efflux transporters,and enhanced DDR and autophagy may be the mechanisms of the resistance of SCLC to EP chemotherapy scheme.
