板蓝根高级不饱和脂肪酸对耐药肝癌细胞株BEL-7404/ADM逆转作用实验

The reverse roles of unsaturated fatty acids from Isatis tinctoria L on human multidrug-resistant hepatocellular carcinoma cell BEL-7404/ADM

目的 :板蓝根高级不饱和脂肪酸对肝癌耐药细胞株BEL 740 4/ADM耐药逆转作用及机制研究。方法 :MTT法观察药物对细胞的生长抑制作用 ;免疫组化法检测细胞P 糖蛋白 (P gp)和多药耐药相关蛋白 (MRP)的表达 ;采用高效液相色谱法(HPLC)测定细胞内ADM的浓度。结果 :耐药肝癌细胞株BEL 740 4/ADM对多种化疗药物产生多药耐药性 ,耐药细胞P gp和MRP的阳性率显著高于亲本细胞 (P <0 .0 1) ;板蓝根高级不饱和脂肪酸在非细胞毒剂量时与ADM合用后能逆转BEL 740 4/ADM对ADM的耐药性 ;ADM与板蓝根高级不饱和脂肪酸合用时 ,其细胞内ADM的含量较单独使用明显增高。结论 :肝癌细胞株BEL 740 4/ADM对多种化疗药物具有多药耐药性 ,其多药耐药性与P gp和MRP过量表达有关 ;板蓝根高级不饱和脂肪酸在非细胞毒剂量时能逆转BEL 740 4/ADM对ADM的耐药性 ,其逆转作用可能与降低P gp药物外排功能。

OBJECTIVE:To study the reverse roles and mechanisms of the unsaturated fatty acids from Isatis tinctoria L on human multidrug-resistant hepatocellular carcinoma cell BEL-7404/ADM. METHOD:BEL-7404/ADM sensitivity to multianticancer agent was evaluated by MTT assay. The expressions of P-gp and MRP were detected by immunohistochemical method. The content of ADM in cells was determined by high performance liquid chromatography (HPLC).RESULTS:the human multidrug -resistant hepatocarcinoma cell line BEL-7404/ADM showed the drug resistance to ADM?MMC and hydroxycamptothecin . The expressions of P-gp and MRP in BEL-7404/ADM were significantly higher than those in BEL-7404 (P<0.01). The unsaturated fatty acids from Isatis tinctoria L in noncytotoxic concentration had reverse role to ADM for BEL-7404/ADM cells. When ADM coupled with the unsaturated fatty acids ,the ADM accumulation in cells increased significantly. (P<0.01) CONCLUSION:BEL-7404/ADM cells is a multidrug resistant model, which multidrug resistance is related with over-expression of P-gp and MRP. The unsaturated fatty acids from Isatis tinctoria L can reverse the resistance of BEL-7404/ADM, and the reverse role may be related with the increasement of anticancer agents content in cells.