Department of Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, 600 Yishan Road, 200233 Shanghai, China.
Mol Biol Rep. 2011 Jun;38(5):3075-82. doi: 10.1007/s11033-010-9975-7. Epub 2010 Jan 29.
In this study, we aim to establish the human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM which can be grow and passaged steadily in 1 μg/ml concentration of adriamycin in appropriate medium. The human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM was established using the method of exposure to medium with adriamycin alternated between high and low concentration with gradually increasing concentration. Furthermore, QBC939 and QBC939/ADM were both treated with adriamycin, mitomycin and vindesine, and then detected by MTT assay, respectively. Growth cycle and intra-cellular concentrations of ADM within cells of each group were determined by flow cytometry. Expression levels of P-glycoprotein were detected by Western bolt and real-time PCR. Results showed that, compared with QBC939, the inhibitive rates of adriamycin, mitomycin and vindesine to QBC939/ADM were lower. Content of ADM in the QBC939/ADM was lower. Western bolt and real time PCR showed that P-glycoprotein in the QBC939/ADM group was over expressed. Therefore, QBC939/ADM was establish and identified as the multi-drug-resistant cell line, which can grow and be passaged steadily in 1 μg/ml concentration adriamycin in appropriate medium. And the multi-drug-resistant character of QBC939/ADM was indicated to be related to the over expression of P-glycoprotein induced by chemotherapeutic drugs.
在本研究中，我们旨在建立人多药耐药性胆管癌细胞系 QBC939/ADM，该细胞系能够在适当的培养基中以 1μg/ml 的阿霉素浓度稳定生长和传代。人多药耐药性胆管癌细胞系 QBC939/ADM 是通过在高低浓度交替的阿霉素培养基中暴露细胞并逐渐增加浓度的方法建立的。此外，用阿霉素、丝裂霉素和长春新碱分别处理 QBC939 和 QBC939/ADM，并通过 MTT 法检测。用流式细胞术检测各组细胞的生长周期和细胞内 ADM 浓度。用 Western blot 和实时 PCR 检测 P-糖蛋白的表达水平。结果表明，与 QBC939 相比，阿霉素、丝裂霉素和长春新碱对 QBC939/ADM 的抑制率较低。QBC939/ADM 中的 ADM 含量较低。Western blot 和实时 PCR 显示 QBC939/ADM 组 P-糖蛋白过度表达。因此，建立并鉴定了 QBC939/ADM 为多药耐药细胞系，该细胞系能够在适当的培养基中以 1μg/ml 的阿霉素浓度稳定生长和传代。并且 QBC939/ADM 的多药耐药特性表明与化疗药物诱导的 P-糖蛋白过度表达有关。
