Tescalcin knockdown inhibits osteogenic differentiation of rat bone mesenchymal stem cells by Wnt/β-catenin signaling pathway

Abstract

Abstract Background Bone marrow-derived mesenchymal stem cells (BMSCs) have the functions of self-renew and differentiating into osteoblasts, adipocytes and chondrocytes, which are regarded as one of the greatest bioscientific achievements in the regenerative medicine field. Tescalcin (TESC), an EF-hand Ca2+ binding protein, plays a vital role in cell proliferation and differentiation. However, what the role of TESC in BMSCs still is unknown. The purpose of this study was to explore the functions. Methods Adenovirus was constructed to decrease the expression of TESC. BML-284 was used to active Wnt/β-catenin signaling pathway. qRT-PCR and western blot was used to detect the expression of mRNA and protein levels. ALP staining and activity were used to detect the change of ALP. ARS staining and quantitative analysis were used to determine the mineralization capacity. Immunofluorescence was used to show the expression of protein. Results Firstly, we found that the mRNA and protein levels of TESC was increased during the osteogenic differentiation. Next, we determined that TESC knockdown inhibited the expression of osteogenic-related genes and decreased the capacity of mineralization. Then, we found that Wnt/β-catenin signaling was inhibited after TESC Knockdown by detecting the Wnt/β-catenin signaling pathway-related protein expression. Afterwards, BML-284 was demonstrated to active Wnt/β-catenin signaling successfully and utilized to rescue the negative osteogenic differentiation of TESC knockdown. Conclusion In summary, our study indicated that TESC knockdown inhibited osteogenic differentiation of BMSCs by Wnt/β-catenin signaling pathway. We supposed that TESC acted in the progress of osteogenic differentiation as a key regulator. We provided a new target for the application of BMSCs in regenerative medicine.