葡萄膜黑色素瘤对化疗药物的敏感性及影响机制的研究

Multidrugs resistance of uveal melanoma in vitro

目的研究探讨葡萄膜黑色素瘤体外化疗对6种药物的敏感性及其影响机制。方法葡萄膜黑色素瘤体外原代与传代培养,抗HMB45免疫组织化学估计肿瘤细胞纯度,MTT法检测对5-氟脲嘧啶、顺铂、噻替哌、阿霉素、长春新碱和足叶乙甙6种化疗药物的敏感性,蛋白印迹法检测耐药基因bcl-2和LRP的表达,分析其表达程度与耐药性的关系。结果体外药敏试验显示6种化疗药物临床常规剂量,对肿瘤细胞的抑制率均不能达到50%,蛋白印迹结果显示bcl-2在体外培养的肿瘤细胞中均有较高表达,而LRP表达在不同类型的肿瘤细胞中不同,表达的高低与耐药程度呈正相关。结论葡萄膜黑色素瘤体外对临床常用6种化疗药物具有较高耐受性,LRP和bcl-2均与其天然耐药性有关,LRP可能起主要作用。

Objective To investigate the machnism of uveal melanocyte resistence to chemical treatment in vitro. Methods Uveal melanoma cells were cultured in vitro. AntiHMB45 depigment immunohistochemical analysis was used to estimate the purity degree of cultured cells. The resistance of uveal melanoma cells to 5-Fu, Cisplatin, Thriotepa, Doxorubicin, Vincristin and Etoposide was tested by MTT, and the expressions of Bcl-2 and LRP were determined by Westen blot. Results The purity of tumor cells was high. According to the half effect inhibiting concentration, each type of uveal melanoma cells showed different resistance to cytotoxicity agents. No chemotherapy drug tested in this experiment could reach 50% inhibiting effect (IC50) at the concentration of clinical usage. The bcl-2 was expressed in all cultured uveal melanoma cells without a correlation to the sensitivity of drug used. The LRP expression was very different among each case with a positive relation to the sensitivity of drug. Conclusion Higher resistance of uveal melanoma to 8 common chemotherapeutic drugs is related to the higher expression of LRP and bcl-2.