肿瘤原代培养物免疫荧光染色法评价化疗药物选择性的研究

Evaluation of chemotherapeutic drug selectivity by using tumor primary culture immunofluorescence staining

目的:探索建立基于肿瘤原代培养物免疫荧光染色评价化疗物选择性和特异性的新方法。方法:利用手术切除的丛状神经纤维瘤(PNF)组织,混合培养原代肿瘤细胞和非肿瘤细胞并使用尼罗替尼(Nilotinib)和伊马替尼(Imatinib)进行处理,测定细胞增殖和毒性。利用特异性免疫荧光抗体染色处理后的细胞,通过Image J软件计数肿瘤细胞和非肿瘤细胞并计算两者比例变化,评价药物效能。结果:药物处理后,细胞增殖的降低和毒性的增加与药物浓度成正比。荧光细胞计数显示肿瘤细胞和非肿瘤细胞数量在药物处理后均减少,但药物效能具有个体差异性。结论:利用特异性免疫荧光染色计数同一组织混合培养的肿瘤细胞和非肿瘤细胞在药物处理后比例变化,可以评价化疗药物的选择性和特异性。

Objective To establish a new method for evaluating the selectivity and specificity of chemotherapeutic drug based on tumor primary culture immunofluorescence staining. Methods： Plexiform neurofibroma tissues cut from patients were used to culture primary tumor cells and non-tumor cells. These cells were treated by nilotinib and imatinib at different concentrations and stained by specific fluorescent antibodies. Proliferation and drug cytotoxicity were detected. The efficacy of drugs was evaluated after counting the proportion of stained tumor cells and non-tumor cells using ImageJ software. ResultsAfter the drug treatment,the reduct-ion in proliferation and increase in cytotoxic effect were directly propeotional to the concentration of the two anticancer drugs. The fluorescent cell count showed that the numbers of both Schwann cells and fibroblasts were reduced after drug treatment, but the effect of drugs was individuaized. Conclusion These results suggest that after drug treatment measuring the change of proportion of tumor cells and non-tumor cells in a mixed culture enables the assessment of the selectivity and specificity of chemotherapeutic drugs.