The effects of ADAM28 shRNA interference vector on the proliferation,differentiation and apoptosis of hu-man periodontal ligament stem cells
Objective:To investigate the effects of disintegrin metalloproteinase 28(ADAM28)shRNA interference vector on the pro-liferation,differentiation and apoptosis of human periodontal ligament stem cells(HPDLSCs)and the related mechanism.Methods:Transfection efficiency of ADAM28 shRNA interference vector was detected 48 h after transfected into HPDLSCs.Cell proliferation was detected by CCK-8 and cell cycle was detected by flow cytometry(FCM).The activity of alkaline phosphatase(ALP)was tested by en-zyme kinetics,and the expression of CAP and Pax9 were tested by Western blot.FCM was used to determine the apoptosis rate of HP-DLSCs,and Western blot was used to detect the expression of Bax and Bcl-2 proteins.Statistical significance was assessed by SPSS-Windows version 21.0 program.Results:The recombinant interference vector PGPU6/GFP/Neo-ADAM28-shRNA was efficiently transfected into HPDLSCs.In the interference vector group,the cell proliferation decreased,and the cell percentage of S phase,G2+M phase and cell proliferation index(PI=S+G2M)in cell cycle decreased,the ALP value decreased,and the expression levels of CAP and Pax9 proteins decreased;the apoptosis rate of HPDLSCs increased,and the expression levels of Bel-2 and Bax proteins increased.Conclusion:ADAM28 shRNA interference vector can restrain the proliferation and differentiation of HPDLSCs,and induce HPDLSCs apoptosis.ADAM28 shRNA interference vector may inhibit the catalytic cracking of metalloproteinase domain and disintegrin-like do-main,and inhibit the promoting cell proliferation of epidermal growth factor(EGF)-like domain,block Notch signal transmission and participate in the negative feedback regulation mechanism of apoptosis.
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