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miR-195 inhibited abnormal activation of osteoblast differentiation in MC3T3-E1 cells via targeting RAF-1

Chen Chao1, Feng Li2, Zhiping Tan2

  • 1Diabetes Center, Institute of Metabolism and Endocrinology, The Second Xiangya Hospital of Central South University, Changsha 410011, PR China.

Experimental Cell Research
|December 4, 2017
PubMed
Abstract

Insights

MicroRNA-195 (miR-195) inhibits abnormal osteoblast differentiation caused by RAF-1 mutations, offering a potential treatment for Noonan syndrome-related bone deformities. This study shows miR-195 targets RAF-1 to regulate cell growth and differentiation.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • RAF-1 mutations, specifically RAF-1L613V, are linked to bone deformities resembling Noonan syndrome.
  • RAF-1L613V is suspected to abnormally activate osteoblast differentiation in MC3T3-E1 cells.

Purpose of the Study:

  • To investigate the role of RAF-1 in bone deformity.
  • To determine if microRNA-195 (miR-195) can inhibit RAF-1L613V-induced bone deformities.
  • To elucidate the mechanism by which miR-195 affects osteoblast differentiation.

Main Methods:

  • Assessed the influence of wild-type RAF-1 (WT) and RAF-1L613V (L613V) on MC3T3-E1 cell viability and differentiation using MTT assays, ALP, and Alizarin Red S staining.
  • Utilized qRT-PCR and western blot analysis to examine gene and protein expression.
  • Investigated the inhibitory effect and mechanism of miR-195 on osteoblast differentiation by targeting RAF-1.

Main Results:

  • RAF-1, particularly RAF-1L613V, abnormally activates BMP-2-induced osteoblast differentiation in MC3T3-E1 cells.
  • miR-195 transfection suppressed L613V-induced cell viability and osteoblast differentiation.
  • miR-195 reduced the expression of key osteogenic genes (Runx2, OSX, ALP, OCN, DLX5) and directly targeted RAF-1 mRNA's 3'-UTR, decreasing RAF-1 expression.

Conclusions:

  • miR-195 inhibits RAF-1 (WT and L613V) induced hyperactive osteoblast differentiation in MC3T3-E1 cells by targeting RAF-1.
  • miR-195 presents potential as a therapeutic agent for RAF-1L613V-induced bone deformities in Noonan syndrome.
Keywords:
BMP-2Osteoblast differentiationRAF-1miR-195

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