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KMID : 1001020090070010027
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Journal of Urologic Oncology
2009 Volume.7 No. 1 p.27 ~ p.33
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Eatablishment of Gemcitabine-resistant Human Bladder Cancer Cell Line and Investigation of the Drug Resistance Mechanism
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Yoon Sang-Jin
Lee Eun-Sik
Byun Seok-Soo
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Abstract
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Purpose: This study was designed to induce gemcitabine-resistant human bladder cancer cell lines and to investigate the mechanism of gemcitabine-resistance by performing a comparative microarray analysis of gene expression between human bladder cancer cells (T24) and its resistant cell lines.
Materials and Methods: We tried to establish gemcitabine resistant sublines from T24 human bladder transitional carcinoma cell line, by continuous exposure to escalating concentrations of gemcitabine starting with 10ng/ml. CCK-8 assay was used to perform cytotoxicity assay. DNA microarray was used for comparative analysis of gene expression of gemcitabine-resistant bladder cancer cell lines. Reverse transcriptase polymerase chain reaction (RT-PCR) was underwent to analyze the results of DNA microarray and formed the basis of estimation of anti-cancer drug resistance.
Results: Three different gemcitabine-resistant cell lines of T24GR1, T24GR5 and T24GR10 were established each exhibiting resistance at the concentration of 1μg/ml, 5μg/ml and 10μg/ml of gemcitabine, respectively. Cytotoxicity assay revealed survival fraction increased in gemcitabine resistant cell lines according to drug concentration. Gene analysis revealed that higher genetic expression of phosphatidylinositide 3-kinase (PI3K) pathway was observed in T24GR1 compared to T24. Genes related to apoptosis pathway and detoxification system were down-regulated in resistant sublines.
Conclusions: Gemcitabine resistant cell lines were established and several resistance mechanism were suggested in our study.
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KEYWORD
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Urinary bladder neoplasm, Drug resistance, Gemcitabine, Microarray analysis
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