膀胱肿瘤BIU-87细胞耐药诱导及逆转的实验研究

目的诱导BIU-87细胞对羟基喜树碱（HCPT）产生耐药,检测多药耐药相关蛋白和交叉耐药,观察维拉帕米（VPM）逆转耐药性的效果.方法用间隙给药法诱导BIU-87细胞对HCPT产生耐药,MTT法测定交叉耐药性,细胞免疫组化染色检测P-糖蛋白（P-gp）、多药耐药相关蛋白（MRP1）、Bcl-2和生存素Survivin,MTT法和流式细胞仪测定VPM逆转耐药性的效果.结果耐药诱导5个月,BIU-87/HCPT细胞对HCPT、长春新碱（VCR）、丝列霉素（MMC）和顺铂（CDDP）的耐药性均有提高,其中HCPT为55倍,VCR为26.88倍,而对阿霉素（ADM）耐药性提高不明显.经检测P-gp和MRP1均为＋＋,Bcl-2和Survivin均为＋＋＋.VPM逆转耐药性有一定效果.结论BIU-87/HCPT细胞具有多药耐药性,对VCR的耐药指数最大,VPM能逆转其耐药性,但逆转效率低于文献报道,这些结果都可能与肿瘤细胞产生多药耐药的机制复杂和BIU-87/HCPT细胞的Bcl-2与Survivin的表达量比P-gp与MRP1高有关.

Drug Resistance Induction and Reversal of Bladder Cancer BIU-87 Cells

Objective To induce BIU-87 cells drug resistance on hydroxycamptothecin（HCPT）,to detect multidrug resistance-associated proteins and drug cross-resistance and to observe the reversed effect of drug resistance of verapamil（VPM）on the BIU-87/HCPT cells.Methods Using hydroxycamptothecin to induce BIU-87 cells by interval-drug-use method,drug resistance on hydroxycamptothecin was made,and then drug cross-resistance on commonly-used antineoplastic was determined by MTT method.The cell immunohistochemical staining assay was used to detect resistance-associated proteins of P-gp,MRP1,Bcl-2 and Survivin.Then MTT assay and flow cytometry was used to determine the reversed effect of drug resistance of VPM on the BIU-87/HCPT cells.Results After five months of drug resistance induction,drug resistance of BIU-87/HCPT cells to HCPT,vincristine（VCR）,mitomycin（MMC）and cisplatin（MMC）were enhanced to different degrees,of which drug resistance to HCPT increase by 55 times and VCR by 26.88 times.However drug resistance to adriamycin（ADM）was not improved obviously.By the cell immunohistochemical staining assay,the positive rate of P-gp and MRP1 was ＋ ＋,however Bcl-2 and Survivin were ＋ ＋ ＋.Reversal of drug resistance of VPM on the BIU-87/HCPT cells was found.Conclusions BIU-87/HCPT cells with multidrug resistance possess the largest drug resistance index on VCR.VPM can reverse the drug resistance of BIU-87/HCPT cells,but the reversal efficiency of VPM is lower than reported in the documents.These results may be related with the complex multidrug resistance mechanisms of tumor cells,and relatively high Bcl-2 and Survivin expression of BIU-87/HCPT cells compared with P-gp and MRP1.