膀胱癌多药耐药细胞株Pumc-91/ADM的建立及其生物学特性评价

Establishment of Human Multidrug-resistant Bladder Cancer Pumc-91/ADM Cell Line and it’s Biological Characteristics Evaluation

目的多药耐药(multidrug-resistance,MDR)是肿瘤治疗的主要障碍。为探讨体外肿瘤MDR的方法,本研究建立膀胱移行上皮癌多药耐药细胞株Pumc-91/ADM,并研究其生物学特性。方法以人膀胱移行上皮癌细胞株Pumc-91为研究对象,应用阿霉素(ADM)浓度梯度递增法体外诱导建立人膀胱移行上皮细胞癌Pumc-91细胞的多药耐药细胞株Pumc-91/ADM。观察细胞的生长规律,用MTT法检测多药耐药性,流式细胞仪检测其生长周期及细胞内药物ADM荧光强度,反转录RT-PCR半定量检测耐药相关基因的mRNA表达量。结果Pumc-91/ADM与亲本细胞Pumc-91相比其生长速度减慢,倍增期延长,细胞异型性明显,巨细胞增多。MTT法检测显示Pumc-91/ADM较Pumc-91细胞对ADM的耐受度提高了10倍,且对氨甲蝶呤、顺铂、表阿霉素、长春新碱均有不同程度的交叉耐药性。流式细胞术检测细胞内荧光强度Pumc-91/ADM较Pumc-91细胞明显降低,细胞周期中S期细胞减少,G1、G2期细胞增多。RT/PCR检测耐药相关基因结果显示耐药细胞株谷胱甘肽-S-转移酶基因表达明显高于亲本细胞。结论耐药株Pumc-91/ADM具有典型的多药耐药特点,可以为膀胱癌的多药耐药研究提供一个较好的体外研究模型。

Objective Muhidrug - resistance （MDR） is considered to be the major obstacle for cancer chemotherapy. In order to study tumor MDR in vitro, we designed this study to establish human multidrug- resistant Bladder cancer pumc -91/ADM cell line and investigate its biological characteristics. Methods MDR cell line （Puree -91/ADM） was induced by wise selection on exposure to in- creasing dose of Adriamycin （ ADM）. Cell growth was measured and multidrug resistance to multi - anticancer agents was evaluated by MTT Assay. Flow cytometry was performed to determine cell cycle and the ADM concentration of cell line. The expression of MDR - relat- ed genes were determined with reverse polymerase chain reaction （RT- PCR）. Results Compared to Pumc -91, the Pumc -91/ADM cell had a prolonged doubling time. The number of cells in S- phase was decreased in Pumc -91/ADM while those in G1 and G2 phase increased. The Pumc- 91/ADM cell was 10 times more resistant to ADM than the Pamc -91 parent. The Pumc- 91/ADM cell exhibited cross - resistance to methotrexate, vincristine, cisplatin, epirubicin. RT - PCR showed that mRNA expression of GST was significantly increased in Pumc -91/ADM. Conclusion Pumc -91/ADM is human multidrug - resistant, and it offers a model with MDR phenotype for the study of MDR in human bladder cancer.