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Lithium chloride enhances cathepsin H expression and BMP-4 degradation in C3H10T1/2 cells.

Kishimoto KN, Itoi E - Biomed Res Int (2013)

Bottom Line: The effect of canonical Wnt/β-catenin signaling on chondrogenic differentiation induced by transfection of BMP4 expressing plasmid was analyzed.Although BMP4 mRNA expression was not affected by LiCl, LiCl decreased BMP4 protein accumulation.These results suggested that cathepsin H is regulated by Wnt/β-catenin signaling and plays an important role in the regulation of BMP4 biological activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Tohoku University School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan.

ABSTRACT
The effect of canonical Wnt/β-catenin signaling on chondrogenic differentiation induced by transfection of BMP4 expressing plasmid was analyzed. Lithium chloride (LiCl) which mimics canonical Wnt/β-catenin signaling was added to cells transfected with BMP4 expressing plasmid. Although BMP4 mRNA expression was not affected by LiCl, LiCl decreased BMP4 protein accumulation. Gene expression analysis exhibited upregulation of cathepsin H by LiCl treatment. Gene silencing of cathepsin H enhanced BMP4 protein accumulation from BMP4 expressing cells. These results suggested that cathepsin H is regulated by Wnt/β-catenin signaling and plays an important role in the regulation of BMP4 biological activity.

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(a) Gene expression profiles of pCAGGS-BMP4 transfected cells cultured in micromass at day 3. (b) Western blot analysis of culture medium for BMP4 at day 3. (c) Immunostaining of micromass for BMP4 at day 3. BMP4 gene expression levels were not significantly altered by LiCl. However, BMP4 protein accumulation in both culture medium and cells was dose dependently decreased by LiCl. Aggrecan and Col2a1 mRNA expression was decreased by LiCl. BMP antagonist, noggin, gremlin 1 and follistatin showed different responses to LiCl. The mRNA expression of cathepsin H was up-regulated by LiCl. **P < 0.01, ***P < 0.001.
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fig3: (a) Gene expression profiles of pCAGGS-BMP4 transfected cells cultured in micromass at day 3. (b) Western blot analysis of culture medium for BMP4 at day 3. (c) Immunostaining of micromass for BMP4 at day 3. BMP4 gene expression levels were not significantly altered by LiCl. However, BMP4 protein accumulation in both culture medium and cells was dose dependently decreased by LiCl. Aggrecan and Col2a1 mRNA expression was decreased by LiCl. BMP antagonist, noggin, gremlin 1 and follistatin showed different responses to LiCl. The mRNA expression of cathepsin H was up-regulated by LiCl. **P < 0.01, ***P < 0.001.

Mentions: The gene expression profiles of C3H10T1/2 cells transfected with pCAGGS-BMP4 (30 μg/cuvette) were analyzed at day 3 (Figure 3(a)). The levels of BMP4 mRNA did not show significant difference by LiCl treatment. Sox9 mRNA expression was significantly suppressed at 25 mM LiCl treatment. Aggrecan and Col2a1 gene were dose-dependently inhibited by LiCl. The expression levels of antagonists against BMPs were also investigated. Noggin mRNA was dose-dependently suppressed. Gremlin 1 expression were not affected by LiCl. The levels of follistatin were increased in dose-dependent manner. The expression levels of cathepsin H was increased in a dose-dependent manner of LiCl.


Lithium chloride enhances cathepsin H expression and BMP-4 degradation in C3H10T1/2 cells.

Kishimoto KN, Itoi E - Biomed Res Int (2013)

(a) Gene expression profiles of pCAGGS-BMP4 transfected cells cultured in micromass at day 3. (b) Western blot analysis of culture medium for BMP4 at day 3. (c) Immunostaining of micromass for BMP4 at day 3. BMP4 gene expression levels were not significantly altered by LiCl. However, BMP4 protein accumulation in both culture medium and cells was dose dependently decreased by LiCl. Aggrecan and Col2a1 mRNA expression was decreased by LiCl. BMP antagonist, noggin, gremlin 1 and follistatin showed different responses to LiCl. The mRNA expression of cathepsin H was up-regulated by LiCl. **P < 0.01, ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3842059&req=5

fig3: (a) Gene expression profiles of pCAGGS-BMP4 transfected cells cultured in micromass at day 3. (b) Western blot analysis of culture medium for BMP4 at day 3. (c) Immunostaining of micromass for BMP4 at day 3. BMP4 gene expression levels were not significantly altered by LiCl. However, BMP4 protein accumulation in both culture medium and cells was dose dependently decreased by LiCl. Aggrecan and Col2a1 mRNA expression was decreased by LiCl. BMP antagonist, noggin, gremlin 1 and follistatin showed different responses to LiCl. The mRNA expression of cathepsin H was up-regulated by LiCl. **P < 0.01, ***P < 0.001.
Mentions: The gene expression profiles of C3H10T1/2 cells transfected with pCAGGS-BMP4 (30 μg/cuvette) were analyzed at day 3 (Figure 3(a)). The levels of BMP4 mRNA did not show significant difference by LiCl treatment. Sox9 mRNA expression was significantly suppressed at 25 mM LiCl treatment. Aggrecan and Col2a1 gene were dose-dependently inhibited by LiCl. The expression levels of antagonists against BMPs were also investigated. Noggin mRNA was dose-dependently suppressed. Gremlin 1 expression were not affected by LiCl. The levels of follistatin were increased in dose-dependent manner. The expression levels of cathepsin H was increased in a dose-dependent manner of LiCl.

Bottom Line: The effect of canonical Wnt/β-catenin signaling on chondrogenic differentiation induced by transfection of BMP4 expressing plasmid was analyzed.Although BMP4 mRNA expression was not affected by LiCl, LiCl decreased BMP4 protein accumulation.These results suggested that cathepsin H is regulated by Wnt/β-catenin signaling and plays an important role in the regulation of BMP4 biological activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Tohoku University School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan.

ABSTRACT
The effect of canonical Wnt/β-catenin signaling on chondrogenic differentiation induced by transfection of BMP4 expressing plasmid was analyzed. Lithium chloride (LiCl) which mimics canonical Wnt/β-catenin signaling was added to cells transfected with BMP4 expressing plasmid. Although BMP4 mRNA expression was not affected by LiCl, LiCl decreased BMP4 protein accumulation. Gene expression analysis exhibited upregulation of cathepsin H by LiCl treatment. Gene silencing of cathepsin H enhanced BMP4 protein accumulation from BMP4 expressing cells. These results suggested that cathepsin H is regulated by Wnt/β-catenin signaling and plays an important role in the regulation of BMP4 biological activity.

Show MeSH
Related in: MedlinePlus