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The Mycoplasma hyorhinis p37 Protein Rapidly Induces Genes in Fibroblasts Associated with Inflammation and Cancer.

Gomersall AC, Phan HA, Iacuone S, Li SF, Parish RW - PLoS ONE (2015)

Bottom Line: The p37 protein at the surface of Mycoplasma hyorhinis cells forms part of a high-affinity transport system and has been found associated with animal and human cancers.This gene activation was principally via the Tlr4 receptor.Blocking the IL6 receptor or inhibiting STAT3 signalling resulted in increased p37-induced gene expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal, Plant and Soil Science, AgriBio, La Trobe University, Melbourne, Victoria, Australia.

ABSTRACT
The p37 protein at the surface of Mycoplasma hyorhinis cells forms part of a high-affinity transport system and has been found associated with animal and human cancers. Here we show in NIH3T3 fibroblasts, p37 rapidly induces the expression of genes implicated in inflammation and cancer progression. This gene activation was principally via the Tlr4 receptor. Activity was lost from p37 when the C-terminal 20 amino acids were removed or the four amino acids specific for the hydrogen bonding of thiamine pyrophosphate had been replaced by valine. Blocking the IL6 receptor or inhibiting STAT3 signalling resulted in increased p37-induced gene expression. Since cancer associated fibroblasts support growth, invasion and metastasis via their ability to regulate tumour-related inflammation, the rapid induction in fibroblasts of pro-inflammatory genes by p37 might be expected to influence cancer development.

No MeSH data available.


Related in: MedlinePlus

The effect of 0.5 μM VIPER and CP7 on p37-induced gene expression in NIH3T3 fibroblasts.Quantitative PCR (qPCR) analysis of NIH3T3 fibroblasts treated with 25 μg ml-1 p37 for 24 hours (black) or pre-treated for 2 hours with VIPER (grey) or the control peptide CP7 (white) prior to 25 μg ml-1 p37-treatment for 24 hours. Significant differences between CP7 or VIPER+p37 treatments and p37 treatment were calculated using ANOVA analysis (+p≤0.05, ++p≤0.01, +++p≤0.001).
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pone.0140753.g002: The effect of 0.5 μM VIPER and CP7 on p37-induced gene expression in NIH3T3 fibroblasts.Quantitative PCR (qPCR) analysis of NIH3T3 fibroblasts treated with 25 μg ml-1 p37 for 24 hours (black) or pre-treated for 2 hours with VIPER (grey) or the control peptide CP7 (white) prior to 25 μg ml-1 p37-treatment for 24 hours. Significant differences between CP7 or VIPER+p37 treatments and p37 treatment were calculated using ANOVA analysis (+p≤0.05, ++p≤0.01, +++p≤0.001).

Mentions: The rapid increases in Angptl4, LIF, Saa3, IL6 and Vcam1 expression in p37 treated fibroblasts suggested the p37 protein is signalling via the toll-like receptor 4 (Tlr4). NIH3T3 fibroblasts possess Tlr4 since 1 μg ml-1 lipopolysaccharide (LPS) treatment for 6 hours resulted in a 28-fold increase in IL6 expression in NIH3T3 fibroblasts [27]. We treated NIH3T3 fibroblasts with 1 μg ml-1 LPS for 24 hours and found a 2-fold and a 12-fold increase in IL6 and Saa3 expression, respectively (data not shown). The Viral Inhibitor Peptide of Tlr4 (VIPER) and its control peptide (CP7) [28] were used to determine whether p37 signals via Tlr4. NIH3T3 fibroblasts were incubated for 24 hours with p37 (25 μg ml-1) or pre-treated for 2 hours with the VIPER or CP7 peptides (0.5 μM) before the addition of p37. The effect of the peptides on the expression levels of the seven genes most strongly induced by p37 was determined (Fig 2). The p37-induced expression of all seven genes was significantly inhibited by VIPER. Although some CP7-induced inhibition occurred, in most cases this was significantly less than the inhibition caused by VIPER. Treatment of NIH3T3 fibroblasts with 0.5 μM VIPER or CP7 alone had no effect on the genes tested, with the exception of Saa3 which was downregulated by 0.5 fold (S8A Fig).


The Mycoplasma hyorhinis p37 Protein Rapidly Induces Genes in Fibroblasts Associated with Inflammation and Cancer.

Gomersall AC, Phan HA, Iacuone S, Li SF, Parish RW - PLoS ONE (2015)

The effect of 0.5 μM VIPER and CP7 on p37-induced gene expression in NIH3T3 fibroblasts.Quantitative PCR (qPCR) analysis of NIH3T3 fibroblasts treated with 25 μg ml-1 p37 for 24 hours (black) or pre-treated for 2 hours with VIPER (grey) or the control peptide CP7 (white) prior to 25 μg ml-1 p37-treatment for 24 hours. Significant differences between CP7 or VIPER+p37 treatments and p37 treatment were calculated using ANOVA analysis (+p≤0.05, ++p≤0.01, +++p≤0.001).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4626034&req=5

pone.0140753.g002: The effect of 0.5 μM VIPER and CP7 on p37-induced gene expression in NIH3T3 fibroblasts.Quantitative PCR (qPCR) analysis of NIH3T3 fibroblasts treated with 25 μg ml-1 p37 for 24 hours (black) or pre-treated for 2 hours with VIPER (grey) or the control peptide CP7 (white) prior to 25 μg ml-1 p37-treatment for 24 hours. Significant differences between CP7 or VIPER+p37 treatments and p37 treatment were calculated using ANOVA analysis (+p≤0.05, ++p≤0.01, +++p≤0.001).
Mentions: The rapid increases in Angptl4, LIF, Saa3, IL6 and Vcam1 expression in p37 treated fibroblasts suggested the p37 protein is signalling via the toll-like receptor 4 (Tlr4). NIH3T3 fibroblasts possess Tlr4 since 1 μg ml-1 lipopolysaccharide (LPS) treatment for 6 hours resulted in a 28-fold increase in IL6 expression in NIH3T3 fibroblasts [27]. We treated NIH3T3 fibroblasts with 1 μg ml-1 LPS for 24 hours and found a 2-fold and a 12-fold increase in IL6 and Saa3 expression, respectively (data not shown). The Viral Inhibitor Peptide of Tlr4 (VIPER) and its control peptide (CP7) [28] were used to determine whether p37 signals via Tlr4. NIH3T3 fibroblasts were incubated for 24 hours with p37 (25 μg ml-1) or pre-treated for 2 hours with the VIPER or CP7 peptides (0.5 μM) before the addition of p37. The effect of the peptides on the expression levels of the seven genes most strongly induced by p37 was determined (Fig 2). The p37-induced expression of all seven genes was significantly inhibited by VIPER. Although some CP7-induced inhibition occurred, in most cases this was significantly less than the inhibition caused by VIPER. Treatment of NIH3T3 fibroblasts with 0.5 μM VIPER or CP7 alone had no effect on the genes tested, with the exception of Saa3 which was downregulated by 0.5 fold (S8A Fig).

Bottom Line: The p37 protein at the surface of Mycoplasma hyorhinis cells forms part of a high-affinity transport system and has been found associated with animal and human cancers.This gene activation was principally via the Tlr4 receptor.Blocking the IL6 receptor or inhibiting STAT3 signalling resulted in increased p37-induced gene expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal, Plant and Soil Science, AgriBio, La Trobe University, Melbourne, Victoria, Australia.

ABSTRACT
The p37 protein at the surface of Mycoplasma hyorhinis cells forms part of a high-affinity transport system and has been found associated with animal and human cancers. Here we show in NIH3T3 fibroblasts, p37 rapidly induces the expression of genes implicated in inflammation and cancer progression. This gene activation was principally via the Tlr4 receptor. Activity was lost from p37 when the C-terminal 20 amino acids were removed or the four amino acids specific for the hydrogen bonding of thiamine pyrophosphate had been replaced by valine. Blocking the IL6 receptor or inhibiting STAT3 signalling resulted in increased p37-induced gene expression. Since cancer associated fibroblasts support growth, invasion and metastasis via their ability to regulate tumour-related inflammation, the rapid induction in fibroblasts of pro-inflammatory genes by p37 might be expected to influence cancer development.

No MeSH data available.


Related in: MedlinePlus