Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2015, Vol. 37 ›› Issue (1): 75-81.doi: 10.3881/j.issn.1000-503X.2015.01.014

• Orginal Article • Previous Articles     Next Articles

Analysis of the Impact of Extracellular Acidity on the Expression and Activity of P-glycoprotein and on the P-glycoprotein-mediated Cytotoxicity of Daunorubicin in Cancer Cell by Microfluidic Chip Technology

Yuan LI, Jiao XIANG, Sha-sha ZHANG, Bei-zhong LIU, Fang GONG, Ming-qing PENG()   

  1. Central Laboratory,Yongchuan Hospital,Chongqing Medical University,Chongqing 402160,China
  • Received:2014-06-09 Online:2015-02-28 Published:2015-02-12
  • Supported by:
    Supported by the Natural Science Foundation Project of CQ CSTC(cstc2012jjA10046),the Medical Science Research Project of Chongqing Health Bureau(2012-2-165),and the District Science and Technology Projects of Yongchuan (YCSTC,2012BE5002)

Abstract: ObjectiveTo explore the impact of extracellular acidic environment on the expression and activity of P-glycoprotein(P-gp)and on the P-gp-mediated cytotoxicity of daunomycin in cancer cells by using microfluidic chip technology. MethodsThe A549 cells cultured on a microfluidic chip were divided into experiment group and control group. The experiment group was exposed to an acidic cell culture medium(pH 6.6),while the control group was treated with a neutral cell culture medium(pH 7.4). The expression of P-gp was detected by cell immunofluorescense analysis and the activity of P-gp was evaluated by Rhodamine 123 efflux experiment. Meanwhile,the cytotoxicity of daunomycin was analyzed by cell live/dead fluorescence staining method. ResultsMicrofluidic chip designed in this study could provide a suitable microenvironment for the growth of A549 cells and the A549 cells reached the confluence of 90% after inoculation for 72 h. Treatment of the acidic cell culture media on A549 cells did not make a significant difference on the expression level of P-gp. However,the activity of P-gp was significantly enhancement and peaked at 6 h after treatment with acidic cell culture media. Meanwhile,the cytotoxicity of daunomycin reduced significantly after treatment with acidic cell culture medium for 6 h,and a reversal effect was obtained when synergy with verapamil. ConclusionsMicrofluidic chip technology can shorten the analysis time and reduce the reagent consumption. It can be used as a new technology platform for understanding the mechanisms of multi-drug resistance and for screening highly efficient multi-drug resistance reversal agents.

Key words: multi-drug resistance, acidic extracellular environment, P-glycoprotein, microfluidic chip, cytotoxicity

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