小细胞肺癌耐药细胞的建立及生物学特性和多药耐药性鉴定  

Establishment and biological characterization of drug-resistant cells and identification of multidrug resistance in small-cell lung cancer

作　　者：韩勇青 王正元 戴秀芬 王子冉 李静[1,2] 戚欣 HAN Yong-qing;WANG Zheng-yuan;DAI Xiu-fen;WANG Zi-ran;LI Jing;QI Xin(School of Medicine,Ocean University of China,Key Laboratory of Marine Medicine,Ministry of Education,Qingdao Shandong 266003,China;Qingdao National Laboratory for Marine Science and Technology-Marine Medicines and Biologics Laboratory,Qingdao Shandong 266237,China)

机构地区：[1]中国海洋大学医药学院,海洋药物教育部重点实验室,山东青岛266003 [2]青岛海洋科学与技术国家实验室-海洋药物与生物制品实验室,山东青岛266237

出　　处：《中国药理学通报》2024年第2期279-284,共6页Chinese Pharmacological Bulletin

基　　金：山东省自然科学基金资助项目(No ZR2023MH116);国家自然科学基金资助项目(No 82273847)。

摘　　要：目的建立对顺铂与依托泊苷抵抗的小细胞肺癌(small cell lung cancer,SCLC)耐药细胞NCI-H446/EP,对其生物学特性和多药耐药性进行评价。方法裸鼠皮下接种SCLC NCI-H446细胞构建异种移植瘤体内模型,给予SCLC一线EP方案治疗,诱导体内耐药,剥离瘤组织体外培养获得耐药细胞。体外实验检测其耐药系数,细胞倍增时间,细胞周期分布,多药耐药基因(MDR1)、耐药相关蛋白的表达,体内验证其耐药性。结果NCI-H446成瘤裸鼠给予EP治疗,8周后产生获得性耐药,经分离培养获得耐药细胞株NCI-H446/EP。该细胞株对顺铂、依托泊苷、SN38和阿霉素的耐药系数分别是12.01、18.36、65.4和10.12。与亲本细胞相比,耐药细胞G 0/G 1期比例明显增加,G 2/M期细胞比例减少,倍增时间延长。耐药细胞MDR1在mRNA及蛋白水平均高于亲本细胞,在体内也具有耐药性。结论成功构建了SCLC耐药细胞株NCI-H446/EP,该细胞具有多药耐药的基本特征,可用于抗SCLC药物筛选及耐药机制的研究。Aim To establish NCI-H446/EP for small cell lung cancer resistant cells resistant to cisplatin and etoposide,and to evaluate their biological characteristics and multidrug resistance.Methods Nude mice were subcutaneously inoculated with NCI-H446 cells of SCLC to construct an in vivo model of xenograft tumor,and were given first-line EP regimen treatment for SCLC,inducing drug resistance in vivo,and stripping tumor tissue in vitro culture to obtain drug-resistant cells.The resistance coefficient,cell doubling time,cell cycle distribution,expression of multidrug resistance gene(MDR1),and drug resistance-related protein were detected in vitro,and the drug resistance to cisplatin and etoposide in vivo were verified.Results Mice with NCI-H446 tumors acquired resistance after eight weeks′EP regimen treatment,and the drug-resistant cell line NCI-H446/EP was obtained by isolation and culture in vitro.The resistance factors of this cell line to cisplatin,etoposide,SN38 and doxorubicin were 12.01,18.36,65.4 and 10.12,respectively.Compared with parental cells,the proportion of NCI-H446/EP cells in G 0/G 1 phase increased significantly,the proportion of G 2/M phase cells decreased,and the doubling time was prolonged.The expressions of MDR1 and drug resistance-related protein of NCI-H446/EP were higher than parental cells.And NCI-H446/EP cells were also resistant to EP regimen treatment in vivo.Conclusions The drug-resistant cell line NCI-H446/EP of small cell lung cancer has been successfully constructed,which has the basic characteristics of multidrug resistance,and can be used for anti-SCLC drug screening and the study of drug resistance mechanism.

关 键 词：小细胞肺癌 化疗 顺铂 依托泊苷 细胞周期 多药耐药 

分 类 号：R-332[医药卫生] R329.28R734.2R979.1