应用软琼脂培养法筛选抗癌药物及获得耐药克隆细胞的研究

Research on Screening of the Antitumor Drugs and Obtaintion of Drug-resistant Clone Cell by Soft Agarose Culture

目的 :探寻可靠的诱导细胞分化剂和筛选有效抗癌药物的最佳剂量及配伍 ,并获得耐药克隆细胞。方法 :向软琼脂糖培养基内添加促分化剂全反式维甲酸 (RA)和 /或免疫制剂抗人Fas抗体 ,在评估不同处理因素下 ,对人胶质母细胞瘤系LN 4 2 8的细胞克隆形成能力影响的同时 ,获得耐药克隆细胞。结果 :在不同处理条件下 ,LN 4 2 8细胞克隆形成能力有明显差异。获得正常对照组和含 10 μmol/LRA的LN 4 2 8克隆细胞 ,在形态学上明显不同。同时含有 10 μmol/LRA +10 0ng/ml抗Fas抗体的培养基内 ,则无细胞克隆形成。 结论 :应用软琼脂培养 ,能有效检测化学制剂对肿瘤细胞的杀伤或促分化效应。探寻针对抗LN 4 2 8肿瘤的最佳药物剂量和配伍 ,同时进一步分离出耐药的克隆细胞 。

Objective: To explore the reliable inducer for cell differentiation, screen the optimal dose and compatible regime of effetive antitumor drugs and obtain the drug-resistant clone cell. Method: RA(the differentiation-facilitated drug) or immune product of the human anti-Fas antibody was added to the soft agarose medium, to evaluate the influence of different disposal on the ability to form LN-428 cell clone in human glioblastoma cell line, and obtain the drug-resistant clones cell simultaneously. Result: The ability to form the LN-428 cell clone is obviously different in different condition. The obviously different result is also acquired between the normal control group and the test group of containing 10 μmol/ L RA LN-428 cell clone in morphology. Meanwhile there is no cell clone forms in the medium which contains 10 μmol/ L RA+100 ng/ ml anti-Fas antibody. Conclusions: The soft agarose culture can be used to test the killing and differentiation-facilitated effect of the chemical drug on the tumor cell. Explortion of the best compatible regime and dose for LN-428 tumor, and furthermore isolation of the drug-resistant cell clone simultaneously provide a new way for experiment oncology and tumor iatreusiology.