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Chinese Journal of Clinical Pharmacology and Therapeutics ›› 2014, Vol. 19 ›› Issue (9): 966-972.

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Infusion receptor eDR4/iFas enhances TRAIL-induced apoptosis of HT-29 cells

LI Zhao-fa1, 2, WANG Cong-yang1, DENG Xiao-ying1, HUI Er-jing1   

  1. 1 School of Biomedical Sciences, Huaqiao University, Quanzhou 362021,Fujian, China;
    2 Molecular Medicine Engineering Research Center of the Ministry of Education, Huaqiao University, Quanzhou 362021, Fujian, China
  • Received:2013-09-23 Revised:2014-06-26 Online:2014-09-26 Published:2014-09-26

Abstract: AIM: To investigate the effect of sTRAIL combined with infusion receptor eDR4/iFas on apoptosis of HT-29 cells. METHODS: In this study, eDR4 and iFas genes were amplified respectively by PCR and used to generate fusion gene named eDR4/iFas by splicing overlap extension PCR. eDR4/iFas was cloned into the expression vector with the tumor-specific survivin promoter, and expressed sTRAIL with constitutive promoter CAG. After treatment with eDR4/iFas and sTRAIL, the cell viability analysis of both colorectal cancer cells HT-29 and human embryonic kidney cells HEK293 was assayed by MTT. Both TRAIL mRNA and protein expressions were detected with RT-PCR and ELISA respectively. The apoptosis of HT-29 cells was analyzed by Hoechst staining. RESULTS: In HT-29 cells, cell survival rate in sTRAIL group was 40.9%±18.7%, while cell survival rate in eDR4/iFas+sTRAIL group was 21.6%±9.1%, there was significant difference between the two groups (P<0.05). CONCLUSION: The results have revealed that eDR4/iFas can improve TRAIL-induced apoptosis of HT-29 cells, has no toxicity for HEK293 cells.

Key words: sTRAIL, infusion receptor, eDR4/iFas, survivin promoter, colorectal cancer cell line HT-29

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