Journal of Shandong University (Health Sciences) ›› 2020, Vol. 58 ›› Issue (3): 87-93.doi: 10.6040/j.issn.1671-7554.0.2019.1367

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MiR-222-5p promotes osteo/odontogenic differentiation of stem cells from human apical papilla

SONG Mengxiao1, WANG Yan2,3,4, LIU Jinzhong1   

  1. 1. Department of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China;
    2. Department of VIP Center, School and Hospital of Stomatology, Shandong University, Jinan 250012, Shandong, China;
    3. Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan 250012, Shandong, China;
    4. Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan 250012, Shandong, China
  • Online:2020-03-10 Published:2022-09-27

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Abstract: Objective To investigate the role of miR-222-5p in osteo/odontogenic differentiation of human stem cells from apical papilla(SCAP)and the related mechanism. Methods The expression of 19 miRNAs in osteo/odontogenic differentiation of SCAP was verified with qPCR. The mRNA expressions of DSPP and Runx2 were measured after SCAP was transfected with 19miRNAs to induce osteo/odontogenic differentiation. The protein expressions of DSPP and Runx2 in the high-expression miR-222-5p group were determined with Western blotting. The potential target gene of miR-222-5p was predicted with bioinformatics analysis. The wild-type(WT)/mutant-type(Mut)reporter vector was constructed to co-transfect HEK293T cells with miR-222-5p. The luciferase activities were evaluated using the dual-luciferase reporter assay system to prove the relationship between miR-222-5p and the target gene. Then the expression of the target gene was tested with Western blotting after overexpression of miR-222-5p. The effects of miR-222-5p and target gene on osteo/odontogenic differentiation of SCAP were further explored. Results The expression of 19 miRNAs detected by qPCR was consistent with that detected by microarray in which miR-222-5p was highly expressed. The mRNA and protein expressions of DSPP and Runx2 were upregulated in miR-222-5p transfected group. Bioinformatics analysis and dual luciferase reporter assay identified SFRP4 as the direct target gene of miR-222-5p, and overexpression of miR-222-5p could inhibit SFRP4 expression. SFRP4 knockdown could effectively reverse the inhibitory effect of miR-222-5p inhibitor on the mRNA expression of DSPP. Conclusion miR-222-5p targetsd SFRP4 and promoted osteo/odontogenic differentiation of SCAP.

Key words: miR-222-5p, Stem cells from apical papilla, Osteo/odontogenic differentiation, SFRP4, Wnt signaling pathway

CLC Number: 

  • R781.3
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