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Published on: August 13, 2013
[Adenovirus containing c-FLIPs gene induces antiapoptosis activation of lymphocyte in vitro]
Zi-yi Feng1, Bao-jin Li, Xiao-ping Liu
1Department of Hepatobillary Surgery, Peking University Shenzhen Hospital, Shenzhen 518036, China.
Objective:
To investigate the efficacy and mechanisms of recombinant adenovirus containing c-FLIPs (cellular FLICE-inhibitory protein short) in inducing antiapoptosis activation of lymphocyte in vitro.
Methods:
c-FLIPs gene was cloned from total RNA of lymphocyte with RT-PCR methods, then constructed recombined adenovirus Ad.c-FLIPs by pAdeasy system. H9 cells infected with Ad.c-FLIPs produced antiapoptosis derived from anti-Apo-1 antibody, and those antiapoptosis evaluated with PI stain flow cytometry (FCM) and Hoechst stain.
Results:
c-FLIPs gene was successfully cloned from lymphocytes and incorporated into recombinant adenovirus Ad.c-FLIPs.After exposure to anti-Apo-1 for 24 h, the apoptosis of H9 cells preconditioned by Ad.c-FLIPs for 24 h as measured by Hochst stain or FCM significantly decreased, compared with non-preconditioned cells, 3.60% +/- 0.21% vs. 48.33% +/- 7.41%, respectively, indicating that Ad.c-FLIPs preconditioning protected H9 cells against apoptosis induced by anti-Apo-1.
Conclusion:
Recombinant adenovirus Ad.c-FLIPs effectively induced anti-apoptosis activation of lymphocyte.
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