Effect of ERRα in silencing DKK1 and SOST recombinant adenovirus vector-transfected MG63 Cells and bone-related proteins
10.16571/j.cnki.1008-8199.2018.09.002
- VernacularTitle: 雌激素相关受体α对沉默重组腺病毒载体转染MG63细胞和骨相关蛋白的影响
- Author:
Shao-jin LIU
1
;
Lei WAN
2
;
Rong-qin QIAO
2
;
Hong-xing HUANG
2
;
Shuang CHAI
1
Author Information
1. Guangzhou University of Chinese Medicine of the Third Clinical Medical School, Guangzhou 510405,Guangdong, China
2. Department of Orthopedics, Guangzhou University of Chinese Medicine at the Third Hospital,Guangzhou 510240,Guangdong, China
- Publication Type:Journal Article
- Keywords:
estrogen receptor-related receptor α；Dickkopf1;
sclerostin；osteoblast；adenovirus vector
- From:
Journal of Medical Postgraduates
2018;31(9):904-909
- CountryChina
- Language:Chinese
-
Abstract:
Objective WNT signaling pathway plays an important role in the formation, differentiation and maturation of bone cells, it is a classical intracellular signaling pathway involved in bone metabolism. DKK1 and Sost play a negative regulatory role in regulating bone mass and osteoblast differentiation, and are negative regulators of WNT signaling pathway. Estrogen-related receptor alpha (ERRα) regulates the functional activity of osteoblasts. The aim of study was to investigate the effect of ERRα on the transfection of MG63 cells and related proteins by the WNT signaling pathway inhibitor Dickkopf (DKK)1 and sclerostin (SOST) adenovirus vectors.Methods The cultured MG63 cells were divided into blank control group, silencing DKK1 group, silencing SOST group, silencing (DKK1+SOST) group, ERRα intervention empty adenovirus group, ERRα intervention silencing DKK1 group, ERRα intervention silencing SOST group, ERRα intervention silencing (DKK1+SOST) group. MG63 cells were transfected with packaged silencing DKK1 and SOST adenovirus vectors according to different groups. The activity of MG63 cells was detected by MTT assay, the activity of ALP was detected by alkaline phosphatase kit, and the concentration of calcium ion was analyzed by flow cytometry. Western blot was used to detect the expressions of low density lipoprotein associated protein 5 (LRP5), bone morphogenetic protein 2 (BMP2), osteopontin (OPN), osteoprotegerin(OPG).Results (1) Compared with blank control group, silencing DKK1, SOST, DKK1+SOST group and ERRα overexpression in the empty adenovirus group could increase cell activity, ALP activity, and decrease calcium ion concentration and increase the expressions of LRP5, BMP2, OPN, and OPG. Differences between groups were statistically significant(P<0.05). (2)Compared with silencing DKK1, silencing SOST, and silencing (DKK1+SOST) groups, the interference of ERRα overexpression with silencing DKK1, silencing SOST, and silencing (DKK1+SOST) groups could also increase cell viability, ALP activity, decrease calcium ion concentration, and as increase LRP5, BMP2, OPN, and OPG expression levels and the differences between groups were statistically significant (P<0.05). However, the interference of ERRα overexpression decreased ALP activity in the silencing SOSt group(P>0.05).Conclusion ERRα Overexpression can increase the activity of MG63 cells, ALP activity, LRP5, BMP2, OPN, and OPG proteins, and decrease the calcium ion concentration in silencing DKK1 and SOST adenovirus-transfected cells.
