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Interleukin-1β-induced autophagy-related gene 5 regulates proliferation of embryonic stem cell-derived odontoblastic cells.

Ozeki N, Hase N, Hiyama T, Yamaguchi H, Kawai R, Kondo A, Matsumoto T, Nakata K, Mogi M - PLoS ONE (2015)

Bottom Line: Our findings suggest that MMP-3 plays a potentially unique physiological role in the generation of odontoblast-like cells under an inflammatory state.IL-1β increased the mRNA and protein levels of Atg5, microtubule-associated protein 1 light chain (LC3, a mammalian homolog of yeast Atg8) and Atg12.Our siRNA analyses combined with western blot analysis revealed a unique sequential cascade involving Atg5, Wnt5a and MMP-3, which resulted in the potent increase in odontoblastic cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, 2-11 Suemori-dori, Chikusa-ku, Nagoya, Aichi, 464-8651, Japan.

ABSTRACT
We previously established a method for the differentiation of induced pluripotent stem cells and embryonic stem cells into α2 integrin-positive odontoblast-like cells. We also reported that Wnt5 in response to interleukin (IL)-1β induces matrix metalloproteinase (MMP)-3-regulated cell proliferation in these cells. Our findings suggest that MMP-3 plays a potentially unique physiological role in the generation of odontoblast-like cells under an inflammatory state. Here, we examined whether up-regulation of autophagy-related gene (Atg) 5 by IL-1β was mediated by Wnt5 signaling, thus leading to increased proliferation of odontoblast-like cells. IL-1β increased the mRNA and protein levels of Atg5, microtubule-associated protein 1 light chain (LC3, a mammalian homolog of yeast Atg8) and Atg12. Treatment with siRNAs against Atg5, but not LC3 and Atg12, suppressed the IL-1β-induced increase in MMP-3 expression and cell proliferation. Our siRNA analyses combined with western blot analysis revealed a unique sequential cascade involving Atg5, Wnt5a and MMP-3, which resulted in the potent increase in odontoblastic cell proliferation. These results demonstrate the unique involvement of Atg5 in IL-1β-induced proliferation of embryonic stem cell-derived odontoblast-like cells.

No MeSH data available.


Related in: MedlinePlus

IL-1β-induced expression of LC3 mRNA and protein in odontoblast-like cells.Two stem cell-derived odontoblast-like cell lines and an odontoblast cell line (KN-3) were treated with IL-1β (0, 0.25, 2.5, and 25 ng/mL). (A–D) qPCR analysis of LC3 mRNA expression relative to the control (18S rRNA). Data are the means ± SD of four independent experiments. **P < 0.01. (E–H) Western blot analysis of LC3 and β-tubulin protein levels following stimulation with IL-1β. Representative blots of three independent experiments are shown.
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pone.0124542.g002: IL-1β-induced expression of LC3 mRNA and protein in odontoblast-like cells.Two stem cell-derived odontoblast-like cell lines and an odontoblast cell line (KN-3) were treated with IL-1β (0, 0.25, 2.5, and 25 ng/mL). (A–D) qPCR analysis of LC3 mRNA expression relative to the control (18S rRNA). Data are the means ± SD of four independent experiments. **P < 0.01. (E–H) Western blot analysis of LC3 and β-tubulin protein levels following stimulation with IL-1β. Representative blots of three independent experiments are shown.

Mentions: It is known that bone-associated cells express other Atgs such as LC3 and Atg12 [36,37]. To assess whether induction of LC3 and Atg12 by IL-1β is a specific response in stem cell-derived odontoblast-like cells, we evaluated the expression of these genes following treatment with the same concentrations of IL-1β (0, 0.25, 2.5 and 25 ng/mL). We found that the mRNA and protein levels of LC3 and Atg12 were also increased by IL-1β at 0.25 and 2.5 ng/mL, but not 25 ng/mL (Figs 2 and 3).


Interleukin-1β-induced autophagy-related gene 5 regulates proliferation of embryonic stem cell-derived odontoblastic cells.

Ozeki N, Hase N, Hiyama T, Yamaguchi H, Kawai R, Kondo A, Matsumoto T, Nakata K, Mogi M - PLoS ONE (2015)

IL-1β-induced expression of LC3 mRNA and protein in odontoblast-like cells.Two stem cell-derived odontoblast-like cell lines and an odontoblast cell line (KN-3) were treated with IL-1β (0, 0.25, 2.5, and 25 ng/mL). (A–D) qPCR analysis of LC3 mRNA expression relative to the control (18S rRNA). Data are the means ± SD of four independent experiments. **P < 0.01. (E–H) Western blot analysis of LC3 and β-tubulin protein levels following stimulation with IL-1β. Representative blots of three independent experiments are shown.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4403923&req=5

pone.0124542.g002: IL-1β-induced expression of LC3 mRNA and protein in odontoblast-like cells.Two stem cell-derived odontoblast-like cell lines and an odontoblast cell line (KN-3) were treated with IL-1β (0, 0.25, 2.5, and 25 ng/mL). (A–D) qPCR analysis of LC3 mRNA expression relative to the control (18S rRNA). Data are the means ± SD of four independent experiments. **P < 0.01. (E–H) Western blot analysis of LC3 and β-tubulin protein levels following stimulation with IL-1β. Representative blots of three independent experiments are shown.
Mentions: It is known that bone-associated cells express other Atgs such as LC3 and Atg12 [36,37]. To assess whether induction of LC3 and Atg12 by IL-1β is a specific response in stem cell-derived odontoblast-like cells, we evaluated the expression of these genes following treatment with the same concentrations of IL-1β (0, 0.25, 2.5 and 25 ng/mL). We found that the mRNA and protein levels of LC3 and Atg12 were also increased by IL-1β at 0.25 and 2.5 ng/mL, but not 25 ng/mL (Figs 2 and 3).

Bottom Line: Our findings suggest that MMP-3 plays a potentially unique physiological role in the generation of odontoblast-like cells under an inflammatory state.IL-1β increased the mRNA and protein levels of Atg5, microtubule-associated protein 1 light chain (LC3, a mammalian homolog of yeast Atg8) and Atg12.Our siRNA analyses combined with western blot analysis revealed a unique sequential cascade involving Atg5, Wnt5a and MMP-3, which resulted in the potent increase in odontoblastic cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, 2-11 Suemori-dori, Chikusa-ku, Nagoya, Aichi, 464-8651, Japan.

ABSTRACT
We previously established a method for the differentiation of induced pluripotent stem cells and embryonic stem cells into α2 integrin-positive odontoblast-like cells. We also reported that Wnt5 in response to interleukin (IL)-1β induces matrix metalloproteinase (MMP)-3-regulated cell proliferation in these cells. Our findings suggest that MMP-3 plays a potentially unique physiological role in the generation of odontoblast-like cells under an inflammatory state. Here, we examined whether up-regulation of autophagy-related gene (Atg) 5 by IL-1β was mediated by Wnt5 signaling, thus leading to increased proliferation of odontoblast-like cells. IL-1β increased the mRNA and protein levels of Atg5, microtubule-associated protein 1 light chain (LC3, a mammalian homolog of yeast Atg8) and Atg12. Treatment with siRNAs against Atg5, but not LC3 and Atg12, suppressed the IL-1β-induced increase in MMP-3 expression and cell proliferation. Our siRNA analyses combined with western blot analysis revealed a unique sequential cascade involving Atg5, Wnt5a and MMP-3, which resulted in the potent increase in odontoblastic cell proliferation. These results demonstrate the unique involvement of Atg5 in IL-1β-induced proliferation of embryonic stem cell-derived odontoblast-like cells.

No MeSH data available.


Related in: MedlinePlus