JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES)

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Targeted killing effects of double suicide genes controlled by the MDR1 promoter on C6/ADR cells

WANG Bo1，WANG Zhi-yun2，WANG Cheng-wei1，WANG Zhi-gang1，ZHANG Yuan1，ZHANG Qing-lin1   

  1. 1. Department of Neurosurgery, Second Hospital of Shandong University;2. Weifang People′s Hospital, Weifang 261041, Shandong, China
  • Received:2007-05-30 Revised:1900-01-01 Online:2008-01-16 Published:2008-01-16
  • Contact: WANG Cheng-wei

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Abstract: Objective To investigate the targeted killing effects of a double suicide gene system controlled by the MDR1 promoter with the prodrug GCV+5-FC on C6/ADR cells. Methods The recombinant vector pcDNA3.MDR1P.CD.TK containing CD and TK double suicide genes controlled by the MDR1 promoter was transfected into C6/ADR cells by using liposome. PCR and RT-PCR were used to identify the integration and expressions of the CD and TK genes. This recombinant vectortransfected C6 cells (C6/CDTK) and normal C6/ADR cells were set as controls. After adding the prodrug, the effects of double suicide genes on cell growth, cell cycle and proliferation were determined by cell growth curve, flow cytometry (FCM) and plate colony formation assay. ResultsPCR proved double suicide genes were integrated into C6/ADR and C6 cells. RT-PCR revealed that the CD and TK genes were expressed in C6/ADR/CDTK cells C6/ADR/CDTK cells was inhibited. G1 phase proportion was significantly higher in C6/ADR/CDTK cells (99.93%) than in C6/ADR cells and C6/CDTK cells (32.68% and 47.57%, P<0.05); the formation rate was significantly lower in C6/ADR/CDTK cells ［(16.7±0.9)%］ than in C6/ADR cells and C6/CDTK cells ［(96.7±2.1)% and (86.7±1.9)%, P<0.05］. ConclusionThe pcDNA3.MDR1P.CDTK/GCV+5FC system has targeted killing effects in C6/ADR cells.

Key words: Multidrug resistance gene, Promoter, Transfection, Glioma, Gene therapy

CLC Number: 

  • R739.41
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