耐顺铂人鼻咽癌细胞系HNE1/CDDP的建立及其生物学特性的研究

Establishment of a cisplatin-induced human nasopharyngeal carcinoma drug-resistant cell line and its biological characteristics

目的:建立耐顺铂(CDDP)的鼻咽癌耐药细胞系HNE1/CDDP,并研究其生物学特性。方法:采用逐步递增培养基中CDDP的方法,体外连续培养建成一株耐CDDP的人鼻咽癌耐药细胞系HNE1/CDDP。用MTT法测定该耐药细胞系对多种抗癌药的敏感性,观察亲本细胞系和耐药细胞系的形态学差异,用生长曲线分别计算亲本细胞系和耐药细胞系的倍增时间,以流式细胞技术检测亲本细胞系和耐药细胞系的周期分布,观测亲本和耐药细胞系黏附、运动和侵袭能力的差别。结果:历时10个月建成人鼻咽癌CDDP耐药细胞系HNE1/CD-DP,能在含CDDP 1mg/L的培养基中稳定生长并传代,其对CDDP的耐药指数为5.83,并与卡铂、奥沙利铂、足叶乙甙等多种抗癌药存在不同程度的交叉耐药性;体外群体倍增时间较亲代细胞延长;细胞周期分析发现其S期与G2/M期细胞减少、G0/G1期细胞增多;HNE1和HNE1/CDDP细胞株的黏附、运动、侵袭能力无明显差别。结论:HNE1/CDDP细胞具有耐药表型,且耐药性稳定,能作为体外研究耐药的模型。

Objective：To establish a CDDP-resistant cell line from human nasopharyngeal carcinoma and evaluate its biological characteristics. Method： By continuously exposing and gradually increasing dose of cisplatin （CDDP）, a resistant nasopharyngeal carcinoma cell line （HNE1/CDDP） was established. Drug sensitivity of this celt line was detected by MTT assay; the alterations of its biological characteristics were determined using light mi- croscopy, cell counting and flow cytometry （FCM） ; its ability of adhesion, migration and invasion were also eval- uated. Result： HNE1/CDDP cell line was developed after 10 months with stable resistance to cisplatin with the resistance index was 5.83. HNE1/CDDP cell exhibited cross resistance to many other chemotherapeutic agents （carboplatin, oxaliplatin and etoposide, etc）. The morphology of HNE1/CDDP changed; doubling time prolonged; and the cell number of S-phase and G2/M-phase decreased while of G0/G1 phase increased compared with parental cells. The ability of adhesion, migration and invasion had no difference between the parental and the resistant cells. Conclusion： HNE1/CDDP cell line shows the typical and stable resistant phenotype and can be used as a research model.