Bone morphogenetic protein 2-induced C2C12 and MC3T3-E1 osteoblast differentiation and autophagy

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Abstract

BACKGROUND:

A series of studies indicate that autophagy is closely linked with differentiation. Bone morphogenetic protein 2 (BMP-2) is the classical pathway for C2C12 and MC3T3-E1 osteoblast differentiation, and the ideal model to study osteogenic differentiation process.

OBJECTIVE:

To observe the relationship between autophagy and BMP-2-induced cell lines C2C12, MC3T3-E1 osteoblast differentiation.

METHODS:

Real-Time PCR was applied to detect osteogenic differentiation and autophagy related index after C2C12 and MC3T3-E1 were induced with BMP-2 (100 μg/L) for 72 hours. The osteogenic index alkaline phosphatase in BMP-2-induced MC3T3-E1 and C2C12 cultured with different concentrations of 3-methyladenine (0, 1, 5, 10 mmol/L) was determined with alkaline phosphatase staining. Western blot analysis was applied to detect LC3-I/II expression levels in C2C12 and MC3T3-E1 induced with BMP-2 for different time points (0, 12, 24, 48, 72, 96 hours).

RESULTS AND CONCLUSION:

The autophagy-related mRNA and protein expression showed an increasing tendency and autophagy-related protein LC3 levels was increased, which was associated with the time, during the BMP-2-induced cell lines C2C12, MC3T3-E1 osteoblast differentiation. Meanwhile, alkaline phosphatase expression levels were inhibited by autophagy in the process of osteogenic differentiation. Therefore, there is a close relationship between autophagy and the BMP-2-induced cell lines C2C12, MC3T3-E1 osteoblast differentiation.

Subject headings: autophagy; bone morphogenetic proteins; cell line; myoblasts; osteoblasts

Funding: the National Natural Science Foundation of China, No. 81171178

Gao Y, Cheng C, Li J, Zhang Y, Xiao WF, Pan QH. Bone morphogenetic protein 2-induced C2C12 and MC3T3-E1 osteoblast differentiation and autophagy. Zhongguo Zuzhi Gongcheng Yanjiu. 2014;18(20):3236-3241.

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