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. 2022 Jan 18;27(3):613.
doi: 10.3390/molecules27030613.

Thyroid-Stimulating Hormone Favors Runx2-Mediated Matrix Mineralization in HOS and SaOS2 Cells: An In Vitro and In Silico Approach

Ramajayam Govindan  1   2 , Mohamed El-Sherbiny  3 , Khalid Mohamed Morsy Ibraheem  4 , Srinivasan Narasimhan  2 , Mohamed El-Dosoky Mohamed Salama  5 , Fazil Ahmad  4 , Selvaraj Jayaraman  6 , Vishnu Priya Veeraraghavan  6 , Srinivasan Vengadassalapathy  7 , Surapaneni Krishna Mohan  8 , Vidhya Rekha Umapathy  9 , Gayathri Rengasamy  6 , Shazia Fathima Jaffer Hussain  10 , Maheshkumar Poomarimuthu  1 , Senthilkumar Kalimuthu  11
Affiliations

Thyroid-Stimulating Hormone Favors Runx2-Mediated Matrix Mineralization in HOS and SaOS2 Cells: An In Vitro and In Silico Approach

Ramajayam Govindan et al. Molecules. .

Abstract

Osteoporosis is a skeletal disease that is both systemic and silent characterized by an unbalanced activity of bone remodeling leading to bone loss. Rising evidences demonstrate that thyroid stimulating hormone (TSH) has an important role in the regulation on the metabolism of bone. However, TSH regulation on human osteoblast essential transcriptional factors has not been identified. Current study examined the role of TSH on human osteoblastic Runx2 expression and their functional genes by in vitro and in slico analysis. Human osteoblast like (HOS and SaoS-2) cells were cultured with DMEM and treated with hTSH at the concentration of 0.01 ng/mL and 10 ng/mL. After treatment, osteoblastic Runx2 and IGF-1R beta expression were studied using RT-PCR and western blot analysis. TSH treatment induced osteoblastic essential transcriptional factor, Runx2 in HOS and SaOS2 cells on 48 h duration and elevated the expression of IGF-IR β gene and Protein in SaoS-2 cells. TSH also promotes Runx2 responsive genes such as ALP, Collagen and osteocalcin in SaOS2 cells on day 2 to day 14 of 10 ng/mL of treatment and favors' matrix mineralization matrix in these cells. In addition, TSH facilitated human osteoblastic cells to mineralize their matrix confirmed by day 21 of alizarin red calcium staining. In silico study was performed to check CREB and ELK1 interaction with Runx2. Results of in silico analysis showed that TSH mediated signalling molecules such as CREB and ELK1 showed interaction with Runx2 which involve in osteobalstic gene expression and differentiation. Present findings confirm that TSH promotes Runx2 expression, osteoblastic responsive genes and bone matrix formation.

Keywords: ALP; CREB; ELK1; Runx2; SaOS2; TSH.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
TSH influences Runx2 expression in HOS cells. Each bar diagram shows mean ± SEM of three experiments; ‘a’ represents compared with control at p < 0.05.
Figure 2
Figure 2
TSH influences Runx2 expression in SaOS-2 cells. Each bar diagram shows mean ± SEM of three experiments; ‘a’ represents compared with control at p < 0.05.
Figure 3
Figure 3
TSH influences expression of IGF-IRB in SaOS-2 cells. Each bar diagram shows mean ± SEM of three experiments; ‘a’ represents compared with control at p < 0.05.
Figure 4
Figure 4
Effect of TSH on Osteocalcin, collagen and ALP mRNA expression in SaOS-2 cells. Each bar diagram shows mean ± SEM of three experiments; ‘a’ represents compared with control at p < 0.05.
Figure 5
Figure 5
Effect of TSH on mineralization in SaOS-2 cells (Phase contrast Microscopy X 10). Each bar diagram shows mean ± SEM of three experiments; ‘a’ represents compared with control at p < 0.05.
Figure 6
Figure 6
The three-dimensional structure of Runx2 using the SWISS-MODEL.
Figure 7
Figure 7
Protein–protein interaction of Runx2 with CREB.
Figure 8
Figure 8
(A,B): Protein–protein interaction of Runx2 with ELK1.
See this image and copyright information in PMC

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References

    1. Abe E., Marians R.C., Yu W., Wu X.B., Ando T., Li Y., Iqbal J., Eldeiry L., Rajendren G., Blair H.C., et al. TSH is a negative regulator of skeletal remodeling. Cell. 2003;115:151–162. doi: 10.1016/S0092-8674(03)00771-2. - DOI - PubMed
    1. Sampath T.K., Simic P., Sendak R., Draca N., Bowe A.E., O’Brien S., Schiavi S.C., McPherson J.M., Vukicevic S. Thyroid-stimulating hormone restores bone volume, microarchitecture, and strength in aged ovariectomized rats. J. Bone Miner. Res. 2007;22:849–859. doi: 10.1359/jbmr.070302. - DOI - PubMed
    1. Sun L., Vukicevic S., Baliram R., Yang G., Sendak R., McPherson J., Zhu L.L., Iqbal J., Latif R., Natrajan A., et al. Intermittent recombinant TSH injections prevent ovariectomy-induced bone loss. Proc. Natl. Acad. Sci. USA. 2008;105:4289–4294. doi: 10.1073/pnas.0712395105. - DOI - PMC - PubMed
    1. Mazziotti G., Sorvillo F., Piscopo M., Cioffi M., Pilla P., Biondi B., Iorio S., Giustina A., Amato G., Carella C. Recombinant human TSH modulates in vivo C-telopeptides of type-1 collagen and bone alkaline phosphatase, but not osteoprotegerin production in postmenopausal women monitored for differentiated thyroid carcinoma. J. Bone Miner. Res. 2005;20:480–486. doi: 10.1359/JBMR.041126. - DOI - PubMed
    1. Reddi A.H., Sullivan N.E. Matrix-induced endochondral bone differentiation: Influence of hypophysectomy, growth hormone, and thyroid-stimulating hormone. Endocrinology. 1980;107:1291–1299. doi: 10.1210/endo-107-5-1291. - DOI - PubMed

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