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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 cisplatin-resistant nasopharyngeal carcinoma cell line HNE1 / CDDP and study its biological characteristics. Methods: A CDDP-resistant human nasopharyngeal carcinoma cell line HNE1 / CDDP was established by continuous culturing in vitro with CDDP method. The sensitivity of the drug-resistant cell line to various anticancer drugs was determined by MTT assay. Morphological differences between the parental cell lines and drug-resistant cell lines were observed. The growth curves were used to calculate the doubling time of the parental and drug- The cell cycle distribution of parental and drug resistant cell lines was detected by flow cytometry to observe the differences of adhesion, motility and invasiveness between parental and drug-resistant cell lines. Results: The human nasopharyngeal carcinoma CDDP resistant cell line HNE1 / CD-DP was established after 10 months and could stably grow and pass on in medium containing 1 mg / L CDDP. The drug resistance index of CDDP was 5.83, Carboplatin, oxaliplatin, etoposide and other anticancer drugs have different degrees of cross-resistance; in vitro population doubling time longer than the parental cells; cell cycle analysis found that the S phase and G2 / M phase cells decreased , G0 / G1 phase cells increased; HNE1 and HNE1 / CDDP cell adhesion, motility, invasion ability no significant difference. Conclusion: The HNE1 / CDDP cells have the drug-resistant phenotype, and the drug resistance is stable, which can be used as a model for the study of drug resistance in vitro.