论文部分内容阅读
目的 在体外进行细胞对抗艾滋病药产生耐性的相关因子研究。方法 通过将成人T细胞白血病细胞系MT-4培养于含有奈非那韦浓度不断增加直至14μmol/L的培养液中培养30天以获得耐奈非那韦的亚系细胞株MT-4rN。用细胞增殖分析系统四唑嗡染色法测定存活细胞数并用流式细胞仪及荧光化合物罗丹明-123和P-糖蛋白阻遏物维拉帕米分析多药耐药蛋白P-gp的存在。结果 发现MT-4rN细胞对新近报道的抗艾滋病药K-37产生出比母本细胞多20倍的耐性。对其它抗肿瘤药的交叉耐性也有不同程度的提高。并且,耐性细胞的荧光密度在维拉帕米存在下有些不同,意味着P-gp在MT-4rN中的表达略有增多。结论 在用HIV-1蛋白酶抑制物奈非那韦作用后,MT-4细胞对其它抗艾滋病药及抗肿瘤药产生了交叉耐性并且多药耐药蛋白P-gp的表达有所增加。
Objective To study the related factors of cell tolerance to AIDS drugs in vitro. Methods MT-4rN sublineline cell line was obtained by culturing adult T-cell leukemia cell line MT-4 in a medium containing nelfinavir at an increasing concentration up to 14 μmol / L for 30 days. The number of surviving cells was determined by tetrazolium staining with the cell proliferation assay system and the presence of the multidrug resistance protein P-gp was analyzed by flow cytometry and the rhodamine-123 and p-glycoprotein repressor verapamil. The results showed that MT-4rN cells produced 20-fold more resistance to the newly reported anti-AIDS drug K-37 than the maternal cells. Cross-resistance to other anti-tumor drugs also have varying degrees of improvement. Moreover, the fluorescent density of the resistant cells is somewhat different in the presence of verapamil, meaning that the expression of P-gp is slightly increased in MT-4rN. CONCLUSION MT-4 cells are cross-resistant to other anti-HIV and antitumor drugs and have an increased expression of the multidrug resistance protein P-gp after treatment with nelfinavir, an HIV-1 protease inhibitor.