[摘要]　目的：建立人大细胞肺癌多药耐药株H460/cDDP，并对其生物学特性进行测定。

方法：以人大细胞肺癌细胞株H460为亲本细胞，采用顺铂大剂量间歇诱导法建立多药耐药细胞株H460/cDDP。光镜下观察其形态学变化；MTT法测定药物敏感性；锥虫蓝拒染实验测定细胞生长曲线、倍增时间；进行染色体核型分析；裸鼠成瘤实验测定其体内成瘤活性。

结果：历时近6个月建成人大细胞肺癌多药耐药株H460/cDDP，经鉴定其对顺铂的耐药指数为10.21，对5-氟尿嘧啶、多柔比星（阿霉素）、依托泊苷（足叶乙甙）、甲氨蝶呤有不同程度的交叉耐药。H460/cDDP较亲本细胞分裂高峰延迟，倍增时间由20.78小时延长至36.46小时，细胞形态改变，染色体变化不突出。经过反复冻存、复苏，上述特性保持稳定，并具有体内成瘤活性，成瘤时间较亲本细胞有所延迟。

结论：新建大细胞肺癌多药耐药株H460/cDDP呈中等度耐药，多药耐药表型稳定且具有体内成瘤活性，可用于下游实验。

[关键词]　肺肿瘤；大细胞肺癌；多药耐药性；顺铂；培养的肿瘤细胞

[Abstract]　Purpose：To establish a human large cell lung cancer multi-drug resistance cell line H460/cDDP and explore its biological characteristics.

Methods：A resistant human larget cell lung cancer cell line (H460/cDDP) was established by intermittent high dose cisplatin selection from the parental cell line H460. Drug sensitivity was detected by MTT assay. The changes of its biological characteristics were determined using light microscopy, Trypan Blue staining rejection, cell counting, chromatosome analysis; Neoplasia formation test in nude mouse was performed to investigate its in vivo characteristic.

Results：H460/cDDP cell line was developed after about 6 months and the resistance index to cisplatin was 10.21. H460/cDDP cells exhibited cross-resistance to 5-Fuorouracil, adriamycn, etoposide and methotrexate. Compared with the parent cells, the morphology wac changed; doubling time prolonged (from 20.78h to 36.46h), while the chromatosome number and caryotype were similar. After being frozen, deposited and resuscitated repeatedly, its biological characteristics remained stable. It had neoplasia formation ability  in vivo, but the forming time was longer than its parental cell.

Conclusions：The newly established multi-drug resistant large cell lung cancer cell line H460/cDDP cell line possessed the typical multi-drug resistant phenotype. It was stable and had neoplasia formation ability in vivo,suitable for research.

[Key words]　lung neoplasm;large cell lung cancer cell Multi-drug resistance;cisplatine; cultured tumor cells



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