Construction Of The Scap Recombinant Adenovirus And Study On The Effects Of The Chondrocytes Proliferation And Apoptosis

Objective:To construct the recombinant adenovirus containing small interfering RNA(siRNA) targeted against human SREBP cleavage-activating protein(SCAP) gene and containing Cricetulus griseus SCAP gene and study its effects on the apoptosis of chondrocyte,which contributes to further investigate the biological function of SCAP gene and provides some reference value for osteoarthritis related to the disrupted lipid metabolism. Methods:By using AdEasyTM adenovirus vector system,to build and pack Ad-SCAP and siSCAP adenoviruses. The ATDC5 cells were infected with Ad-SCAP and siSCAP adenoviruses and the expression levels of SCAP mRNA and protein were detected by RT-PCR and western blot respectively. The effects of SCAP recombinant adenovirus on the proliferation and apoptosis of ATDC5 cells were detected by FCM and western blot..Results:The recombinant adenovirus Ad-SCAP and siSCAP were constructed successfully. When compared with control groups(NC、Ad-GFP/Ad-RFP), the apoptosis rate of ATDC5 cells infected with Ad-SCAP was increased(P<0.05) but decreased(P<0.05) when infected with si SCAP adenovirus. similarly, the proliferation rate was decreased significantly(P<0.05) when infected with Ad-SCAP adenovirus and increased(P<0.05) when infected with siSCAP. Western blotting results of cleaved Caspase-3 and p-JNK were in accordance with those results shown by FCM.Conclusion:Infecting ATDC5 cells with Ad-SCAPsiRNA promotes the proliferation and suppresses the apoptosis in the cells; Infecting ATDC5 cells with Ad-SCAP suppresses the proliferation and promotes the apoptosis in the cells.