Study on cell proliferation and apoptosis by human ATF6 gene adenovirus modification in vitro

Abstract:

Objective：To construct the adenovirus vector containing human activating transcription factor 6（ATF6） gene and to study the effects on the proliferation and apoptosis of SW1353 cells in the stress condition. Methods：The ATF6 gene and the siRNA se－quence targeting ATF6 gene were respectively cloned into the shuttle plasmid pAdTrack-cmv and pSES-HUS，and then were ho－mogenously recombined with the adenovirus backbone plasmid pAdEasy-l in Ecoli BJ5183 to generate the adenovirus vector pAd-ATF6 and pAd-ATF6 siRNA.The recombinant adenovirus vector was transfected into the 293 packing cells by lipofectamine-mediated transfection to amplify the recombinant adenovirus Ad-ATF6 and Ad-ATF6 siRNA. The SW1353 cells were infected with the aden－ovirus and the expression levels of ATF6 mRNA and protein were detected by RT-PCR and Western blot. The effects of the recombi－nant adenovirus on the proliferation and apoptosis of SW1353 in the stress condition were detected by FCM，MTT and Western blot. Results：The recombinant adenovirus Ad-ATF6 and Ad-ATF6 siRNA with high infection were successfully obtained. FCM analysis showed that under stress condition，the S phase of SW1353 cells in the Ad-ATF6 group was increased significantly（P=0.000），and the apoptosis rate was decreased significantly（P=0.000） as compared with those of control group. Meanwhile the results of siATF6 groups were on the control. The MTT results and Western blot results were in accordance with that of FCM. Conclusion：Under stress condition，ATF6 may promote the proliferation of SW1353 cells and suppress its apoptosis.