Construction Of Adenovirus Containing Human ATF6Gene And Effects On The Apoptosis Of Cartilage Cells

Objective：To construct the recombinant adenovirus containing human ATF6gene and study the effects on the apoptosis of cartilage cells. It willcontribute to further study the biological function of ATF6gene andprovide some reference value for cartilage development diseases related toERS.Methods：By using AdEasyTM adenovirus vector system, to build Ad-ATF6and siATF6adenoviruses. The ATDC5cells were infected with theadenoviruses and the expression levels of ATF6mRNA and protein weredetected by RT-PCR and western blot. The effects of Ad-ATF6on theproliferation and apoptosis of ATDC5and C28I2cells were detected byFCM. The effects of siATF6on the proliferation and apoptosis of SW1353in the stress condition were detected by FCM、MTT and Western blot.The effects of Ad-ATF6and siATF6on the apoptosis of ATDC5andC3H10T1/2cells which were induced by BMP2were detected by FCM、IHC、TUNEL and Western blot. Results:The recombinant adenovirus Ad-ATF6and siATF6with highinfection were successfully obtained. The apoptosis rate of ATDC5andC28I2cells infected with Ad-ATF6were increased significantly（P＜0.05），and the rate of reproduction were decreased significantly（P＜0.05）as compared with the control group. The apoptosis rate of TM+siATF6group of SW1353cells was significantly higher than TM+Ad-ATF6group（P＜0.05）. The apoptosis rates of ATDC5、C3H10T1/2cells andcartilage tissue which were induced by BMP2and Ad-ATF6weresignificantly higher than that induced by BMP2and siATF6.Conclusion：Under normal circumstances，the proliferation of cartilage cells couldbe suppressed by Ad-ATF6, and the apoptosis could be promoted. While，in the stress condition，the results were on the contrary. When the Cartilagecells were under differentiation induced by BMP2，ATF6could promotethe ER stress-mediated apoptosis.