胃癌细胞系SGC7901耐阿霉素和顺铂细胞系的建立及其耐药可持续性的质控研究  

Establishment of adriamycin-resistant and cisplatin-resistant gastric cancer cell lines and assessment on their sustainability of drug resistance

作　　者：杜静平 陈鹏 费洪新 杨明慧 陈正华 

机构地区：[1]云南大学生命科学院 [2]甘肃省亚盛集团博士后科研工作站北京分站,北京100101

出　　处：《胃肠病学和肝病学杂志》2007年第4期368-372,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金资助项目(30672063)

摘　　要：目的建立胃癌耐药细胞模型,并进行耐药细胞耐药可持续性研究。方法以脉冲式诱导方法诱导胃癌耐阿霉素细胞SGC7901/ADR和胃癌耐顺铂细胞SGC7901/DDP。检测细胞的生长曲线和群体倍增时间,电镜观察细胞的超微结构,并进行了体外药物敏感性实验和细胞药物转运实验,Western blot检测耐药细胞内P-gp和MRP的表达,尤其是进行了耐药细胞的耐药可持续性的质控研究。结果耐药细胞的增殖能力比药敏细胞强(P<0.05);电镜下耐药细胞较药敏细胞表现为更多的核大核畸形及细胞膜间突触样联系和细胞间小管结构;体外药物敏感性实验和细胞药物转运实验显示耐药细胞的耐药性明显增强;而且耐药细胞中MRP和P-gp的表达均比药敏细胞SGC7901高;耐药可持续性的质控研究表明在不给与抗肿瘤药维持状况下细胞自第11代耐药性明显下降。结论成功构建了胃癌耐阿霉素细胞系SGC7901/ADR和耐顺铂细胞系SGC7901/DDP;对实验用耐药细胞系建议质控在9代以内以保证耐药细胞的耐药稳定性。Objectives To establish the drug-resistant cell models of gastric cancer and assess their sustainahility of drug resistance. Methods Adriamycin-resistant （SGC7901/ADR） and cisplatin-resistant （SGC7901/DDP） gastric cancer cell lines were produced using pulse induction method. The growth curve and population double times were investigated on naive gastric cancer cell line and drug-resistant derivatives. Uhrastructures of cells were examined under an electronic microscope. In vitro MTT assay for drug sensitivity and cell-based assay for drug transport were carried out for the three kinds of cells. The expression levels of P-gp and MRP in drug-resistant cell lines were detected using Western blot. More importantly, the sustainahility of drug-resistant cell lines was carefully assessed. Results Significant difference was found in proliferative capacity between naive gastric cancer cell line （SGC7901） with drug-resistant derivatives （SGC7901/ADR and SGC7901/DDP,P 〈0.05）. Under an electronic microscope, the drug-resistant cell lines exhibited more nuclear enlargement or abnormalities besides intermemhrane synapse-like junction and intercellular tubules, compared with SGC7901 cells. Both in vitro MTT assay for drug sensitivity and cell-based assay for drug transport revealed that the drug resistance was increased in drug-resistant cell lines. Moreover, the expressions of MRP and P-gp were both upregulated in drug-resistant cell lines. It was also found that without the persistent pressure of antitumor drugs, the SGC7901/ADR and SGC7901/DDP cell lines remarkably reduced their drug resistance since the 11 passage. Conclusion We successfully established adriamycin-resistant （SGC7901/ADR） and cisplatin-resistant （SGC7901/ DDP） gastric cancer cell lines. In addition, we suggest here that for experimental use the two drug-resistant cell lines should he raised within 9 passages in order to ensure their drug resistance.

关 键 词：胃癌 多药耐药 耐阿霉素胃癌细胞系 耐顺铂胃癌细胞系 

分 类 号：R735.2[医药卫生—肿瘤]