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Runx2/miR-3960/miR-2861 Positive Feedback Loop Is Responsible for Osteogenic Transdifferentiation of Vascular Smooth Muscle Cells.

Xia ZY, Hu Y, Xie PL, Tang SY, Luo XH, Liao EY, Chen F, Xie H - Biomed Res Int (2015)

Bottom Line: However, the effect of this feedback loop on the osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs) remains unclear.Our recent study showed that miR-2861 and miR-3960 expression increases significantly during β-glycerophosphate-induced osteogenic transdifferentiation of VSMCs.MiR-2861 or miR-3960 promotes osteogenic transdifferentiation of VSMCs by targeting histone deacetylase 5 or Homeobox A2, respectively, resulting in increased runt-related transcription factor 2 (Runx2) protein production.

View Article: PubMed Central - PubMed

Affiliation: Institute of Endocrinology and Metabolism, Second Xiang-Ya Hospital of Central South University, Changsha, Hunan 410011, China.

ABSTRACT
We previously reported that Runx2/miR-3960/miR-2861 regulatory feedback loop stimulates osteoblast differentiation. However, the effect of this feedback loop on the osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs) remains unclear. Our recent study showed that miR-2861 and miR-3960 expression increases significantly during β-glycerophosphate-induced osteogenic transdifferentiation of VSMCs. Overexpression of miR-2861 or miR-3960 in VSMCs enhances β-glycerophosphate-induced osteoblastogenesis, whereas inhibition of miR-2861 or miR-3960 expression attenuates it. MiR-2861 or miR-3960 promotes osteogenic transdifferentiation of VSMCs by targeting histone deacetylase 5 or Homeobox A2, respectively, resulting in increased runt-related transcription factor 2 (Runx2) protein production. Furthermore, overexpression of Runx2 induces miR-2861 and miR-3960 transcription, and knockdown of Runx2 attenuates β-glycerophosphate-induced miR-2861 and miR-3960 transcription in VSMCs. Thus, our data show that Runx2/miR-3960/miR-2861 positive feedback loop plays an important role in osteogenic transdifferentiation of VSMCs and contributes to vascular calcification.

No MeSH data available.


Related in: MedlinePlus

Inhibition of miR-2861 or miR-3960 inhibited β-glycerophosphate-induced osteoblast differentiation of vascular smooth muscle cells. ALP activity and osteocalcin secretion (a) and Western blot analysis of Runx2 protein expression (b) in 10 mM β-glycerophosphate- (β-GP-) treated VSMCs transfected with anti-miR-C (control) or anti-miR-2861, anti-miR-C, or anti-miR-3960. Data are shown as mean ± s.e.m. ∗P < 0.05.
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fig3: Inhibition of miR-2861 or miR-3960 inhibited β-glycerophosphate-induced osteoblast differentiation of vascular smooth muscle cells. ALP activity and osteocalcin secretion (a) and Western blot analysis of Runx2 protein expression (b) in 10 mM β-glycerophosphate- (β-GP-) treated VSMCs transfected with anti-miR-C (control) or anti-miR-2861, anti-miR-C, or anti-miR-3960. Data are shown as mean ± s.e.m. ∗P < 0.05.

Mentions: We next determined the role of miR-2861 or miR-3960 during osteogenic transdifferentiation of VSMCs by changing the functional levels of miR-2861 or miR-3960 in VSMCs. Northern blot data confirmed overexpression of miR-2861 and miR-3960 in VSMCs stably transfected with miR-2861 precursor and miR-3960 precursor, respectively (Figure 2(a)). The transfected VSMCs were treated with β-glycerophosphate for 48 h for inducing osteogenesis. Then, ALP activity, osteocalcin secretion, and Runx2 protein expression, as markers of osteoblast differentiation, were evaluated. The levels of ALP, osteocalcin, and Runx2 were all elevated in cells with overexpression of miR-2861 or miR-3960 (Figure 2(b)). These results indicated that overexpression of miR-2861 or miR-3960 promoted osteogenic transdifferentiation of VSMCs (Figures 2(b) and 2(c)). In contrast, knockdown of miR-2861 or miR-3960 reduced ALP, osteocalcin, and Runx2 levels induced by β-glycerophosphate (Figures 3(a) and 3(b)). All of these results suggest that both miR-2861 and miR-3960 act to promote osteogenic transdifferentiation of VSMCs.


Runx2/miR-3960/miR-2861 Positive Feedback Loop Is Responsible for Osteogenic Transdifferentiation of Vascular Smooth Muscle Cells.

Xia ZY, Hu Y, Xie PL, Tang SY, Luo XH, Liao EY, Chen F, Xie H - Biomed Res Int (2015)

Inhibition of miR-2861 or miR-3960 inhibited β-glycerophosphate-induced osteoblast differentiation of vascular smooth muscle cells. ALP activity and osteocalcin secretion (a) and Western blot analysis of Runx2 protein expression (b) in 10 mM β-glycerophosphate- (β-GP-) treated VSMCs transfected with anti-miR-C (control) or anti-miR-2861, anti-miR-C, or anti-miR-3960. Data are shown as mean ± s.e.m. ∗P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Inhibition of miR-2861 or miR-3960 inhibited β-glycerophosphate-induced osteoblast differentiation of vascular smooth muscle cells. ALP activity and osteocalcin secretion (a) and Western blot analysis of Runx2 protein expression (b) in 10 mM β-glycerophosphate- (β-GP-) treated VSMCs transfected with anti-miR-C (control) or anti-miR-2861, anti-miR-C, or anti-miR-3960. Data are shown as mean ± s.e.m. ∗P < 0.05.
Mentions: We next determined the role of miR-2861 or miR-3960 during osteogenic transdifferentiation of VSMCs by changing the functional levels of miR-2861 or miR-3960 in VSMCs. Northern blot data confirmed overexpression of miR-2861 and miR-3960 in VSMCs stably transfected with miR-2861 precursor and miR-3960 precursor, respectively (Figure 2(a)). The transfected VSMCs were treated with β-glycerophosphate for 48 h for inducing osteogenesis. Then, ALP activity, osteocalcin secretion, and Runx2 protein expression, as markers of osteoblast differentiation, were evaluated. The levels of ALP, osteocalcin, and Runx2 were all elevated in cells with overexpression of miR-2861 or miR-3960 (Figure 2(b)). These results indicated that overexpression of miR-2861 or miR-3960 promoted osteogenic transdifferentiation of VSMCs (Figures 2(b) and 2(c)). In contrast, knockdown of miR-2861 or miR-3960 reduced ALP, osteocalcin, and Runx2 levels induced by β-glycerophosphate (Figures 3(a) and 3(b)). All of these results suggest that both miR-2861 and miR-3960 act to promote osteogenic transdifferentiation of VSMCs.

Bottom Line: However, the effect of this feedback loop on the osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs) remains unclear.Our recent study showed that miR-2861 and miR-3960 expression increases significantly during β-glycerophosphate-induced osteogenic transdifferentiation of VSMCs.MiR-2861 or miR-3960 promotes osteogenic transdifferentiation of VSMCs by targeting histone deacetylase 5 or Homeobox A2, respectively, resulting in increased runt-related transcription factor 2 (Runx2) protein production.

View Article: PubMed Central - PubMed

Affiliation: Institute of Endocrinology and Metabolism, Second Xiang-Ya Hospital of Central South University, Changsha, Hunan 410011, China.

ABSTRACT
We previously reported that Runx2/miR-3960/miR-2861 regulatory feedback loop stimulates osteoblast differentiation. However, the effect of this feedback loop on the osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs) remains unclear. Our recent study showed that miR-2861 and miR-3960 expression increases significantly during β-glycerophosphate-induced osteogenic transdifferentiation of VSMCs. Overexpression of miR-2861 or miR-3960 in VSMCs enhances β-glycerophosphate-induced osteoblastogenesis, whereas inhibition of miR-2861 or miR-3960 expression attenuates it. MiR-2861 or miR-3960 promotes osteogenic transdifferentiation of VSMCs by targeting histone deacetylase 5 or Homeobox A2, respectively, resulting in increased runt-related transcription factor 2 (Runx2) protein production. Furthermore, overexpression of Runx2 induces miR-2861 and miR-3960 transcription, and knockdown of Runx2 attenuates β-glycerophosphate-induced miR-2861 and miR-3960 transcription in VSMCs. Thus, our data show that Runx2/miR-3960/miR-2861 positive feedback loop plays an important role in osteogenic transdifferentiation of VSMCs and contributes to vascular calcification.

No MeSH data available.


Related in: MedlinePlus