Limits...
Interleukin-1 Receptor Antagonist Has a Novel Function in the Regulation of Matrix Metalloproteinase-13 Expression.

Goto H, Ishihara Y, Kikuchi T, Izawa A, Ozeki N, Okabe E, Kamiya Y, Ozawa Y, Mizutani H, Yamamoto G, Mogi M, Nakata K, Maeda H, Noguchi T, Mitani A - PLoS ONE (2015)

Bottom Line: These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA.Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization.In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, School of Dentistry, Aichi Gakuin University, Chikusa-ku, Nagoya, Aichi, Japan.

ABSTRACT
Interleukin-1 receptor antagonist (IL-1Ra) is an IL-1 family member, which binds to IL-1 receptors but does not induce any intracellular signaling. We addressed whether IL-1Ra has a novel function in regulation of the extracellular matrix or adhesion molecules. Polymerase chain reaction array analysis demonstrated a ~5-fold increase in matrix metalloproteinase 13 (MMP-13) mRNA expression of IL-1Ra siRNA-transfected Ca9-22 human oral squamous epithelial carcinoma cells compared with the control. In fact, MMP-13 mRNA and protein expression as well as its activity in IL-1Ra siRNA-transfected Ca9-22 cell lines were significantly higher than those in the control. IL-1Ra siRNA treatment resulted in strong elevation of MMP-13 expression, whereas addition of rhIL-1Ra (40 ng/ml) suppressed MMP-13 expression, suggesting that IL-1Ra had a specific effect on MMP-13 induction. IL-1Ra siRNA could potently suppress IL-1α. No significant difference was found between the MMP-13 mRNA expression of IL-1Ra siRNA-transfected cells and those treated with anti-IL-1α or anti-IL-1β antibodies. These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA. Histopathological investigation of MMP-13 in periodontal tissue showed specific localization in the junctional epithelial cells of IL-1Ra knockout (KO) mice. Furthermore, infection with Aggregatibacter actinomycetemcomitans to establish an experimental periodontitis model resulted in predominant localization of MMP-13 along apical junctional epithelial cells. Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization. In particular, laminin-5 localization almost disappeared in the internal basal lamina of IL-1Ra KO mice infected with A. actinomycetemcomitans, suggesting that the suppression of IL-1Ra resulted in strong induction of MMP-13 that degraded laminin-5. In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

No MeSH data available.


Related in: MedlinePlus

TIMP expression in IL-1Ra siRNA-transfected cells.(A) TIMP-1 and TIMP-2 mRNA expression in IL-1Ra and control siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (B) Western blot of TIMP-1 (23 kDa) and -2 (21 kDa) in cells transfected with either IL-1Ra or control siRNAs. Data are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4608771&req=5

pone.0140942.g004: TIMP expression in IL-1Ra siRNA-transfected cells.(A) TIMP-1 and TIMP-2 mRNA expression in IL-1Ra and control siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (B) Western blot of TIMP-1 (23 kDa) and -2 (21 kDa) in cells transfected with either IL-1Ra or control siRNAs. Data are representative of three independent experiments.

Mentions: MMP-13 activity is precisely regulated after its secretion at the post-translational level as a precursor zymogen and by TIMPs [21, 22]. Although it is known that TIMP-2 is inducible by cytokines [21], we found that TIMP-1 and TIMP-2 mRNA levels were constitutively expressed in IL-1Ra and control siRNA-transfected cells at 1, 3, 6, 12, and 24 hours (Fig 4A). These data were supported by the gene array results shown in Fig 1. To normalize the increase in MMP activity, it is necessary to describe it in the context of TIMP protein expression. As shown in Fig 1, TIMP-1 (23 kDa) and TIMP-2 (21 kDa) proteins were constitutively expressed at uniform levels under all experimental conditions (Fig 4B).


Interleukin-1 Receptor Antagonist Has a Novel Function in the Regulation of Matrix Metalloproteinase-13 Expression.

Goto H, Ishihara Y, Kikuchi T, Izawa A, Ozeki N, Okabe E, Kamiya Y, Ozawa Y, Mizutani H, Yamamoto G, Mogi M, Nakata K, Maeda H, Noguchi T, Mitani A - PLoS ONE (2015)

TIMP expression in IL-1Ra siRNA-transfected cells.(A) TIMP-1 and TIMP-2 mRNA expression in IL-1Ra and control siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (B) Western blot of TIMP-1 (23 kDa) and -2 (21 kDa) in cells transfected with either IL-1Ra or control siRNAs. Data are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608771&req=5

pone.0140942.g004: TIMP expression in IL-1Ra siRNA-transfected cells.(A) TIMP-1 and TIMP-2 mRNA expression in IL-1Ra and control siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (B) Western blot of TIMP-1 (23 kDa) and -2 (21 kDa) in cells transfected with either IL-1Ra or control siRNAs. Data are representative of three independent experiments.
Mentions: MMP-13 activity is precisely regulated after its secretion at the post-translational level as a precursor zymogen and by TIMPs [21, 22]. Although it is known that TIMP-2 is inducible by cytokines [21], we found that TIMP-1 and TIMP-2 mRNA levels were constitutively expressed in IL-1Ra and control siRNA-transfected cells at 1, 3, 6, 12, and 24 hours (Fig 4A). These data were supported by the gene array results shown in Fig 1. To normalize the increase in MMP activity, it is necessary to describe it in the context of TIMP protein expression. As shown in Fig 1, TIMP-1 (23 kDa) and TIMP-2 (21 kDa) proteins were constitutively expressed at uniform levels under all experimental conditions (Fig 4B).

Bottom Line: These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA.Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization.In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, School of Dentistry, Aichi Gakuin University, Chikusa-ku, Nagoya, Aichi, Japan.

ABSTRACT
Interleukin-1 receptor antagonist (IL-1Ra) is an IL-1 family member, which binds to IL-1 receptors but does not induce any intracellular signaling. We addressed whether IL-1Ra has a novel function in regulation of the extracellular matrix or adhesion molecules. Polymerase chain reaction array analysis demonstrated a ~5-fold increase in matrix metalloproteinase 13 (MMP-13) mRNA expression of IL-1Ra siRNA-transfected Ca9-22 human oral squamous epithelial carcinoma cells compared with the control. In fact, MMP-13 mRNA and protein expression as well as its activity in IL-1Ra siRNA-transfected Ca9-22 cell lines were significantly higher than those in the control. IL-1Ra siRNA treatment resulted in strong elevation of MMP-13 expression, whereas addition of rhIL-1Ra (40 ng/ml) suppressed MMP-13 expression, suggesting that IL-1Ra had a specific effect on MMP-13 induction. IL-1Ra siRNA could potently suppress IL-1α. No significant difference was found between the MMP-13 mRNA expression of IL-1Ra siRNA-transfected cells and those treated with anti-IL-1α or anti-IL-1β antibodies. These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA. Histopathological investigation of MMP-13 in periodontal tissue showed specific localization in the junctional epithelial cells of IL-1Ra knockout (KO) mice. Furthermore, infection with Aggregatibacter actinomycetemcomitans to establish an experimental periodontitis model resulted in predominant localization of MMP-13 along apical junctional epithelial cells. Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization. In particular, laminin-5 localization almost disappeared in the internal basal lamina of IL-1Ra KO mice infected with A. actinomycetemcomitans, suggesting that the suppression of IL-1Ra resulted in strong induction of MMP-13 that degraded laminin-5. In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

No MeSH data available.


Related in: MedlinePlus