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目的 :建立人膀胱癌耐阿霉素细胞系及研究它们的生物学特性及药物耐受性的机制。 方法 :采用人膀胱癌细胞系 EJ,经递增阿霉素剂量的方法 ,历时一年 ,建立一株耐药亚株 EJ/ DOR,对其生物学特性及耐药机制进行了研究 ,并应用反转录 PCR检测了 MDR1、MRP和 DNA Topo 基因的表达。 结果 :EJ/ DOR对阿霉素的相对耐受度较亲本细胞提高了 14.3倍 ;对蒽环类、长春花属生物碱及 DNA Topo 靶制剂足叶乙甙有明显的交叉耐药性 ,但对顺铂、丝裂霉素无明显的交叉耐受性 ;耐药细胞对柔红霉素的细胞内聚集量显著减少 ;EJ/ DOR并不表达 MDR1基因 ,而 MRP基因过表达 ,但细胞内 DNATopo 基因表达低于亲本细胞。 结论 :细胞内 DNA Topo 基因表达下降及 MRP基因过表达是 EJ/ DOR表现为多耐药受性亚型的主要原因 ,这种非 P- gp介导的非经典型多耐药受性细胞为寻求包括阿霉素在内的化疗方案提供了良好的实验模型。
OBJECTIVE: To establish human adriamycin resistant human bladder cancer cell lines and to study their biological characteristics and mechanisms of drug tolerance. Methods: Human bladder cancer cell line EJ was used to increase the dose of doxorubicin. After one year, a drug resistant subtype EJ / DOR was established and its biological characteristics and drug resistance mechanism were studied. Transcriptional PCR was used to detect the expression of MDR1, MRP and DNA Topo genes. RESULTS: The relative resistance of EJ / DOR to doxorubicin was 14.3 times higher than that of parental cells. There was a significant cross-resistance to anthracycline, vinca alkaloids and DNA Topo target agent etoposide, There was no obvious cross-tolerance to cisplatin and mitomycin; the intracellular accumulation of daunorubicin in drug-resistant cells was significantly reduced; EJ / DOR did not express MDR1 gene but MRP gene was overexpressed, but intracellular DNATopo gene expression is lower than the parental cells. CONCLUSION: The decrease of Topo gene expression and the over-expression of MRP gene are the main reasons for EJ / DOR expression in MDR subtypes. The non-P-gp mediated multi-drug resistant multi-drug resistant cells are Seeking a chemotherapy protocol that includes doxorubicin provides a good experimental model.