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目的 建立肿瘤耐药机制研究及筛选抗肿瘤药物的体外实验模型。方法 应用药物连续作用并逐步提高药物浓度的剂量递增法及软琼脂细胞克隆技术 ,用拒染法测定药物的细胞毒作用 ,建立了一株对顺铂 (DDP)具有 40倍耐药性的L12 10细胞亚系 (L12 10 /DDP40 )。结果 DDP耐药细胞亚系在倍增时间、集落形成率、细胞周期、DNA指数和细胞形态等方面基本保持了亲代细胞 (L12 10 )的生物学特性。耐药细胞亚系在加药状态下冻存半年后复苏其耐药性仍然存在 ,解除药物作用后 5mon其耐药性仍维持原有水平。耐药细胞亚系对卡铂、丝裂霉素、噻替派、甲氨蝶呤、长春新碱和氮芥具有不同程度的交叉耐药性 ,对三尖杉酯碱和阿霉素无交叉耐药性 ,对阿糖胞苷和氟尿嘧啶仍较敏感。结论 DDP耐药性L12 10细胞亚系的建立 ,为深入研究肿瘤细胞耐药机制、寻找逆转耐药的措施提供了较好的体外实验模型。
Objective To establish an in vitro experimental model for studying the mechanism of tumor drug resistance and screening antitumor drugs. Methods A dose-escalation method using continuous drug and gradually increasing the drug concentration and soft agar cell cloning technique were used. The cytotoxicity of the drug was determined by the method of rejection, and a L12 with 40-fold resistance to cisplatin (DDP) was established. 10 cell sublines (L12 10 /DDP40). Results The biological characteristics of the parental cell (L12 10 ) in the DDP-resistant cell subfamily were basically maintained in terms of doubling time, colony formation rate, cell cycle, DNA index and cell morphology. The drug-resistant cell sub-family revived after being stored for half a year in the dosing state and its drug resistance persisted. After the drug was released, its drug resistance remained at its original level. Drug-resistant cell sublines have varying degrees of cross-resistance to carboplatin, mitomycin, thiotepa, methotrexate, vincristine, and nitrogen mustard, and no crossover to harringtonine and doxorubicin. Resistance is still sensitive to cytarabine and fluorouracil. Conclusion The establishment of D12 resistant L12 10 cell subline provides a good in vitro experimental model for further study of the drug resistance mechanism of tumor cells and finding measures to reverse drug resistance.