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| In vitro and in vivo Gene Therapy Research of CDTK Genes Drove by Enhanced Tumor-specific Promoter in Liver Cancer |
| XUE Jin-feng, XUE Zhi-gang, CHEN Yi-yao, LI Zhuo, YIN Biao, WU Ling-qian, LIANG De-sheng |
| State Key Laboratory of Medical Genetics, Central South University, Changsha 410078, China |
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Abstract Objective: To investigate the in vivo and in vitro killing effects of CDTK gene drove by enhanced hTERT promoter in liver cancer cell line Bel-7402. Methods: Cloned CMV enhancer, hTERT promoter and CDTK gene into hrDNA targeting vector pHrn and constructed pHr-CeTpCDTK-GFP vector. After transferred into Bel-7402 cells, CDTK gene expression and in vitro killing effects were detected by RT-PCR, HPLC and MTT. Finally, the pHr-CeTpCDTK-GFP vector mixed with liposome were injected directly into the tumor of nude mouse, which was produced by the injection of those liver cancer cells, meanwhile, transfected cells were injected into mouse, the tumor size and killing effects were closely observed. Results: The RT-PCR and HPLC results showed that the liver cancer cells transfected with the constructed pHr-CeTpCDTK-GFP vector could express CDTK successfully. The MTT showed that the vector is toxic to the liver cancer cell. Through the animal test, it was found that the concentration of 5-Fu was 7.694μg/ml in treatment group, the tumor volume of treatment group decreased 6.5 times compared with the control group. Using vector-transfected cells induce tumor, the tumor formation time of treatment group was 8 days later than control group, while the mean survival period of treatment group was 16 days longer than control group. Conclusions: The pHr-CeTpCDTK-GFP vector could kill the liver cancer efficiently and resulted in a new approach for the liver cancer gene therapy.
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Received: 03 March 2015
Published: 25 June 2015
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