JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2017, Vol. 55 ›› Issue (3): 43-48.doi: 10.6040/j.issn.1671-7554.0.2016.1068

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Post-transcriptional regulation mechanism of sclerostin in mice

LI Shanshan1,2, YANG Jing1,2, ZHANG Jin1,2,3   

  1. 1.School of Stomatology, Shandong University;
    2. Shandong Provincial Key Laboratory of Oral Tissue Regeneration;
    3. Department of Endodontics, School and Hospital of Stomatology, Shandong University, Jinan 250012, Shandong, China
  • Received:2016-08-29 Online:2017-03-10 Published:2017-03-10

Abstract: Objective To investigate whether the expression of Sost gene was post-transcriptionally regulated in primary osteoblasts and MC3T3-E1 murine pre-osteoblast cells. Methods Osteogenic differentiation was induced in MC3T3-E1 cells and primary calvarial osteoblasts were isolated from C57BL/6 mice. The mRNA and protein levels of Sost gene in these cells were evaluated respectively. The 3'UTR of mouse Sost gene was amplified and subcloned into the SacI restriction site of pMIR-REPORT miRNA expression reporter, and the resulted plasmid was named as pMIR-REPORT-SOST 3' untranslated regions(UTR). Calvarial osteoblasts, MC3T3-E1 cells and NIH3T3 cells were then transfected with pMIR-REPORT-SOST UTR and pMIR-REPORT, respectively. Results Sost mRNA levels in calvarial osteoblasts and MC3T3-E1 cells were higher than control groups(P<0.05)during osteogenic differentiation. In contrast, the protein levels of Sost gene had no significant difference(P>0.05)with control group in calvarial osteoblasts and MC3T3-E1 cells. Relative luciferase levels transfected with pMIR-REPORT-SOST UTR had dramatically decline with transfected with pMIR-REPORT, respectively(P<0.05). In contrast, relative luciferase levels had no statistically significant differences between cells transfected with pMIR-REPORT-SOST UTR and pMIR-REPORT in NIH3T3 cells(P>0.05). Conclusion pMIR-REPORT-SOST UTR was successfully generated, and the expression of Sost gene was post-transcriptionally regulated via the 3'UTR.

Key words: Sclerostin, Osteogenic differentiation, Wnt signaling, Post-transcriptional regulation, 3'Untranslated regions(UTR)

CLC Number: 

  • R574
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