Effect of adseverin on proliferation, migration and mineralization of MDPC-23 cells

Abstract

Objective To explore the effect of adseverin on proliferation, migration and mineralization of MDPC-23 cells. Methods The recombinant lentivirus vector was constructed to knockdown adseverin and transfected into MDPC-23 cells. Cells transfected with non-targeting shRNA or without shRNA (mock transfection) were used as negative control. Western blot was applied to detect the expression of adseverin. Cell proliferation was examined by CCK-8 method and the migration ability was assayed by Transwell. Alkaline phosphatase activity and Alizarin red staining assay were used to test the effect of adseverin on the mineralization of MDPC-23 cells. The level of significance was determined by One-Way ANOVA followed by LSD-t test for a multiple comparison procedure. Rusults Adseverin expression was significantly deceased compared with the control. Silencing adseverin in MDPC-23 significantly hindered cell proliferation by approximately 26.8% (F48 h= 11.025, P = 0.01) , 26.5% (F72 h = 86.205, P < 0.001) followed by 48 h and 72 h of culture in growth medium. The number of cells transferred through the Transwell membrane in shRNA-Adseverin group was dramatically reduced by 61.2% (F = 6.005, P = 0.008) when compared with control. Cells transfected by adseverin shRNA displayed higher level of ALP activity (ALPshRNA-Ads= 568.43 U/gprot, ALPshRNA-Ctrl= 142.56 U/gprot, ALPMDPC-23= 118.16 U/gprot; F= 49.002, P<0.001) and mineralized nodule formation. Conclusion Adseverin might play a crucial role in the proliferation, migration and mineralization in MDPC-23 cells. Key words: Adseverin; MDPC-23; Cell proliferation; Cell movement; Mineralization