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Secretion of protein disulphide isomerase AGR2 confers tumorigenic properties

View Article: PubMed Central - PubMed

ABSTRACT

The extracellular matrix (ECM) plays an instrumental role in determining the spatial orientation of epithelial polarity and the formation of lumens in glandular tissues during morphogenesis. Here, we show that the Endoplasmic Reticulum (ER)-resident protein anterior gradient-2 (AGR2), a soluble protein-disulfide isomerase involved in ER protein folding and quality control, is secreted and interacts with the ECM. Extracellular AGR2 (eAGR2) is a microenvironmental regulator of epithelial tissue architecture, which plays a role in the preneoplastic phenotype and contributes to epithelial tumorigenicity. Indeed, eAGR2, is secreted as a functionally active protein independently of its thioredoxin-like domain (CXXS) and of its ER-retention domain (KTEL), and is sufficient, by itself, to promote the acquisition of invasive and metastatic features. Therefore, we conclude that eAGR2 plays an extracellular role independent of its ER function and we elucidate this gain-of-function as a novel and unexpected critical ECM microenvironmental pro-oncogenic regulator of epithelial morphogenesis and tumorigenesis.

Doi:: http://dx.doi.org/10.7554/eLife.13887.001

No MeSH data available.


Related in: MedlinePlus

A volcano plot representation of differentially expressed gene in comparison of cultures growing in absence or presence of eAGR2.The log Fold Change of the gene expression levels observed in the two conditions is reported on the X axis, while the –log 10 of the p-value of the statistical test of differential expression (t-test) is shown on the Y axis. The horizontal black dotted line represents the threshold of statistical significance of 0.05 for the p-value (1.3 for the -log10 p-value). Genes having Log Fold Change below that threshold are considered not statistically significant (gray dots). Genes with p-value equal or greater than the threshold are considered biologically down-regulated (green dots), up-regulated (red dots) or not biologically significant (orange dots) if their Fold Change is respectively less than or equal to 0.5, greater than or equal to +2 and between 0.5 and 2 (Log Fold Change <= -1, >= +1 and between -1 and +1).DOI:http://dx.doi.org/10.7554/eLife.13887.016
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fig9s1: A volcano plot representation of differentially expressed gene in comparison of cultures growing in absence or presence of eAGR2.The log Fold Change of the gene expression levels observed in the two conditions is reported on the X axis, while the –log 10 of the p-value of the statistical test of differential expression (t-test) is shown on the Y axis. The horizontal black dotted line represents the threshold of statistical significance of 0.05 for the p-value (1.3 for the -log10 p-value). Genes having Log Fold Change below that threshold are considered not statistically significant (gray dots). Genes with p-value equal or greater than the threshold are considered biologically down-regulated (green dots), up-regulated (red dots) or not biologically significant (orange dots) if their Fold Change is respectively less than or equal to 0.5, greater than or equal to +2 and between 0.5 and 2 (Log Fold Change <= -1, >= +1 and between -1 and +1).DOI:http://dx.doi.org/10.7554/eLife.13887.016

Mentions: Since AGR2 has been correlated as potentially involved in EMT (Mizuuchi et al., 2015; Ma et al., 2015) and that EMT is characterized by a reduction of cell-cell adhesion and loss of apico-basolateral polarity (Thiery, 2002), we examined whether eAGR2 might regulate EMT in non-tumorigenic HBEC organoids. To this end, HBEC organoids growth with eAGR2 for 10 days or not, RNA was harvested and analysed using RT-qPCR array. The expression of 84 EMT genes was quantified and presented as a Volcano plot (Figure 9—figure supplement 1). The addition of eAGR2 to the organoids ECM significantly modified EMT transcripts such as Wnt5b (~thirteen-fold), MMP3 (~eleven-fold), MAP1B (~ten-fold), MMP9 (~eight-fold), ZEB1 (~three-fold), and VIM (~two-fold) (Figure 9A). Changes were also observed in genes related to ECM and cell adhesion, cell growth and proliferation, differentiation and development, migration and mobility, and cytoskeleton (Figure 9B). These results were also confirmed at the protein level with the induction of Vimentin (VIM), MMP9 and N-Cadherin (CDH2) expression upon treatment with eAGR2 as well as decreased E-Cadherin (CDH1) expression (Figure 9C). The impact of eAGR2 on EMT induction is also reinforced by the relocalization of E-Cadherin and β-catenin from the basal side of the organoids to the cytoplasm (Figure 8A and B).10.7554/eLife.13887.015Figure 9.Extracellular AGR2 promotes EMT and invasive structures.


Secretion of protein disulphide isomerase AGR2 confers tumorigenic properties
A volcano plot representation of differentially expressed gene in comparison of cultures growing in absence or presence of eAGR2.The log Fold Change of the gene expression levels observed in the two conditions is reported on the X axis, while the –log 10 of the p-value of the statistical test of differential expression (t-test) is shown on the Y axis. The horizontal black dotted line represents the threshold of statistical significance of 0.05 for the p-value (1.3 for the -log10 p-value). Genes having Log Fold Change below that threshold are considered not statistically significant (gray dots). Genes with p-value equal or greater than the threshold are considered biologically down-regulated (green dots), up-regulated (red dots) or not biologically significant (orange dots) if their Fold Change is respectively less than or equal to 0.5, greater than or equal to +2 and between 0.5 and 2 (Log Fold Change <= -1, >= +1 and between -1 and +1).DOI:http://dx.doi.org/10.7554/eLife.13887.016
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Related In: Results  -  Collection

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fig9s1: A volcano plot representation of differentially expressed gene in comparison of cultures growing in absence or presence of eAGR2.The log Fold Change of the gene expression levels observed in the two conditions is reported on the X axis, while the –log 10 of the p-value of the statistical test of differential expression (t-test) is shown on the Y axis. The horizontal black dotted line represents the threshold of statistical significance of 0.05 for the p-value (1.3 for the -log10 p-value). Genes having Log Fold Change below that threshold are considered not statistically significant (gray dots). Genes with p-value equal or greater than the threshold are considered biologically down-regulated (green dots), up-regulated (red dots) or not biologically significant (orange dots) if their Fold Change is respectively less than or equal to 0.5, greater than or equal to +2 and between 0.5 and 2 (Log Fold Change <= -1, >= +1 and between -1 and +1).DOI:http://dx.doi.org/10.7554/eLife.13887.016
Mentions: Since AGR2 has been correlated as potentially involved in EMT (Mizuuchi et al., 2015; Ma et al., 2015) and that EMT is characterized by a reduction of cell-cell adhesion and loss of apico-basolateral polarity (Thiery, 2002), we examined whether eAGR2 might regulate EMT in non-tumorigenic HBEC organoids. To this end, HBEC organoids growth with eAGR2 for 10 days or not, RNA was harvested and analysed using RT-qPCR array. The expression of 84 EMT genes was quantified and presented as a Volcano plot (Figure 9—figure supplement 1). The addition of eAGR2 to the organoids ECM significantly modified EMT transcripts such as Wnt5b (~thirteen-fold), MMP3 (~eleven-fold), MAP1B (~ten-fold), MMP9 (~eight-fold), ZEB1 (~three-fold), and VIM (~two-fold) (Figure 9A). Changes were also observed in genes related to ECM and cell adhesion, cell growth and proliferation, differentiation and development, migration and mobility, and cytoskeleton (Figure 9B). These results were also confirmed at the protein level with the induction of Vimentin (VIM), MMP9 and N-Cadherin (CDH2) expression upon treatment with eAGR2 as well as decreased E-Cadherin (CDH1) expression (Figure 9C). The impact of eAGR2 on EMT induction is also reinforced by the relocalization of E-Cadherin and β-catenin from the basal side of the organoids to the cytoplasm (Figure 8A and B).10.7554/eLife.13887.015Figure 9.Extracellular AGR2 promotes EMT and invasive structures.

View Article: PubMed Central - PubMed

ABSTRACT

The extracellular matrix (ECM) plays an instrumental role in determining the spatial orientation of epithelial polarity and the formation of lumens in glandular tissues during morphogenesis. Here, we show that the Endoplasmic Reticulum (ER)-resident protein anterior gradient-2 (AGR2), a soluble protein-disulfide isomerase involved in ER protein folding and quality control, is secreted and interacts with the ECM. Extracellular AGR2 (eAGR2) is a microenvironmental regulator of epithelial tissue architecture, which plays a role in the preneoplastic phenotype and contributes to epithelial tumorigenicity. Indeed, eAGR2, is secreted as a functionally active protein independently of its thioredoxin-like domain (CXXS) and of its ER-retention domain (KTEL), and is sufficient, by itself, to promote the acquisition of invasive and metastatic features. Therefore, we conclude that eAGR2 plays an extracellular role independent of its ER function and we elucidate this gain-of-function as a novel and unexpected critical ECM microenvironmental pro-oncogenic regulator of epithelial morphogenesis and tumorigenesis.

Doi:: http://dx.doi.org/10.7554/eLife.13887.001

No MeSH data available.


Related in: MedlinePlus