Carbohydrate responsive element binding protein (ChREBP) negatively regulates osteoblast differentiation via protein phosphatase 2A Cα dependent manner

被引:1
作者
Kim, Kyeong-Min [1 ,2 ]
Kim, Eun-Jung [2 ,3 ]
Jang, Won-Gu [1 ,2 ]
机构
[1] Daegu Univ, Sch Engn, Dept Biotechnol, Gyeongbuk 38453, South Korea
[2] Daegu Univ, Res Inst Antiaging, Gyeongbuk 38453, South Korea
[3] Kyungpook Natl Univ, Dept Immunol, Sch Med, Daegu 41944, South Korea
基金
新加坡国家研究基金会;
关键词
ChREBP; PP2A Ca; Osteoblast differentiation; BMP2; Ethanol; FATTY-ACID SYNTHESIS; BONE-FORMATION; GLUCOSE-HOMEOSTASIS; INSULIN-RESISTANCE; GROWTH-FACTORS; EXPRESSION; ETHANOL; ACTIVATION; ALCOHOL; CELLS;
D O I
10.1016/j.biocel.2020.105766
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Carbohydrate responsive element binding protein (ChREBP) is a major transcription factor of lipogenesis regulated by glucose status in the liver. However, the function of ChREBP in osteogenic differentiation is unclear. The present study examined the role of ChREBP in osteoblast differentiation in MC3T3-E1 preosteoblast cell line. The mRNA expression of ChREBP, protein phosphatase 2A catalytic subunit-alpha (PP2A C alpha) and the osteogenic genes such as, DNA-binding protein inhibitor (Idl), runt-related transcription factor-2 (Runx2), and alkaline phosphatase (ALP) was measured by qPCR and RT-PCR. Runx2, ChREBP, and PP2A C alpha, protein levels were evaluated by Western blotting. ALP staining experiment was carried out to evaluate ALP enzyme activity, and a luciferase reporter assay was performed to analyze Runx2 transcriptional activity. Expression of ChREBP and PP2A C alpha did not change during bone morphogenetic protein-2 (BMP2)-induced osteoblast differentiation. Overexpression of ChREBP reduced the osteogenic genes (Runx2 and ALP) expression and ALP activity, while knockdown of ChREBP had the opposite effects. Overexpression of PP2A C alpha increased ChREBP expression, while inhibition of PP2A C alpha using okadaic acid not only inhibited the expression of ChREBP, but also restored the mRNA and protein expression of Runx2 and activity of ALP enzyme. These results demonstrate that ChREBP inhibits BMP2-induced osteoblast differentiation in a PP2A C alpha- dependent manner.
引用
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页数:9
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