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Epithelial-mesenchymal transition markers and HER3 expression are predictors of elisidepsin treatment response in breast and pancreatic cancer cell lines.

Teixidó C, Marés R, Aracil M, Ramón y Cajal S, Hernández-Losa J - PLoS ONE (2013)

Bottom Line: Interestingly, we observed that the basal protein expression levels of EMT markers show a significant correlation with cell viability in response to elisidepsin treatment in a panel of 12 different breast and pancreatic cancer cell lines.In addition, we generated three elisidepsin treatment-resistant cell lines (MCF-7, HPAC and AsPC-1) and analyzed the pattern of expression of different EMT markers in these cells, confirming that acquired resistance to elisidepsin is associated with a switch to the EMT state.These results show that HER3 expression is an important marker of sensitivity to elisidepsin treatment.

View Article: PubMed Central - PubMed

Affiliation: Molecular Pathology Group, Vall d'Hebron Research Institute, Universidad Autonoma of Barcelona, Barcelona, Spain.

ABSTRACT
Elisidepsin (elisidepsin trifluoroacetate, Irvalec®, PM02734) is a new synthetic depsipeptide, a result of the PharmaMar Development Program that seeks synthetic products of marine origin-derived compounds. Elisidepsin is a drug with antiproliferative activity in a wide range of tumors. In the present work we studied and characterized the mechanisms associated with sensitivity and resistance to elisidepsin treatment in a broad panel of tumor cell lines from breast and pancreas carcinomas, focusing on different factors involved in epithelial-mesenchymal transition (EMT) and the use of HER family receptors in predicting the in vitro drug response. Interestingly, we observed that the basal protein expression levels of EMT markers show a significant correlation with cell viability in response to elisidepsin treatment in a panel of 12 different breast and pancreatic cancer cell lines. In addition, we generated three elisidepsin treatment-resistant cell lines (MCF-7, HPAC and AsPC-1) and analyzed the pattern of expression of different EMT markers in these cells, confirming that acquired resistance to elisidepsin is associated with a switch to the EMT state. Furthermore, a direct correlation between basal HER3 expression and sensitivity to elisidepsin was observed; moreover, modulation of HER3 expression levels in different cancer cell lines alter their sensitivities to the drug, making them more resistant when HER3 expression is downregulated by a HER3-specific short hairpin RNA and more sensitive when the receptor is overexpressed. These results show that HER3 expression is an important marker of sensitivity to elisidepsin treatment.

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Expression of EMT markers associated with elisidepsin sensitivity in breast cancer cell lines.Protein expression levels of different EMT markers were evaluated by immunocytochemistry (A), western blot (B) and IHC (C). A) Immunocytochemistry of two epithelial (E-cadherin and β-catenin) and four mesenchymal markers (vimentin, Slug, Snail and Twist). Magnification 100x. B) E-cadherin, β-catenin, Slug, Snail, Twist, vimentin and β-actin (loading control) were detected by western blot analysis using 50 µg of total protein. C) Basal levels of E-cadherin, β-catenin and vimentin were analyzed by IHC. Magnification 20x. Each experiment was performed at least in duplicate.
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pone-0053645-g002: Expression of EMT markers associated with elisidepsin sensitivity in breast cancer cell lines.Protein expression levels of different EMT markers were evaluated by immunocytochemistry (A), western blot (B) and IHC (C). A) Immunocytochemistry of two epithelial (E-cadherin and β-catenin) and four mesenchymal markers (vimentin, Slug, Snail and Twist). Magnification 100x. B) E-cadherin, β-catenin, Slug, Snail, Twist, vimentin and β-actin (loading control) were detected by western blot analysis using 50 µg of total protein. C) Basal levels of E-cadherin, β-catenin and vimentin were analyzed by IHC. Magnification 20x. Each experiment was performed at least in duplicate.

Mentions: We aimed to determine whether the various elisidepsin-sensitive cancer cell lines shared similar basal levels of EMT genes. In the breast cancer cell lines we found E-cadherin expression in the sensitive cell lines. All cell lines had detectable expression of β-catenin, whereas Slug expression was variable and not related to their sensitivity to elisidepsin. Furthermore, Snail expression was only found in MDA-MB-435, and all the cell lines that exhibited levels of Twist-1 and vimentin were less sensitive to the drug (Figs. 2A–C). In contrast, elisidepsin-sensitive pancreatic carcinoma cell lines expressed E-cadherin and β-catenin, whereas the less sensitive cells expressed Slug. Lastly, Snail, Twist-1 and vimentin expression was found in sensitive and insensitive cell lines alike (Figs. 3A–C). To summarize, E-cadherin protein was significantly expressed in the sensitive cell lines independently of their tumoral origin (Mann-Whitney test: p  = 0.0364; Fig. S2), and vimentin was significantly expressed in the less sensitive ones (Mann-Whitney test: p  = 0.0364). On the other hand, Twist-1 and Snail proteins were found in all less sensitive cell lines (Mann Whitney test: p  = 0.0636 and p  = 0.1000, respectively), with the exception of two sensitive cell lines that were positive for vimentin expression (CFPAC and AsPC-1), one sensitive cell line that was positive for Twist-1 expression (CFPAC) and another one that was positive for Snail expression (SKBR3).


Epithelial-mesenchymal transition markers and HER3 expression are predictors of elisidepsin treatment response in breast and pancreatic cancer cell lines.

Teixidó C, Marés R, Aracil M, Ramón y Cajal S, Hernández-Losa J - PLoS ONE (2013)

Expression of EMT markers associated with elisidepsin sensitivity in breast cancer cell lines.Protein expression levels of different EMT markers were evaluated by immunocytochemistry (A), western blot (B) and IHC (C). A) Immunocytochemistry of two epithelial (E-cadherin and β-catenin) and four mesenchymal markers (vimentin, Slug, Snail and Twist). Magnification 100x. B) E-cadherin, β-catenin, Slug, Snail, Twist, vimentin and β-actin (loading control) were detected by western blot analysis using 50 µg of total protein. C) Basal levels of E-cadherin, β-catenin and vimentin were analyzed by IHC. Magnification 20x. Each experiment was performed at least in duplicate.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3539985&req=5

pone-0053645-g002: Expression of EMT markers associated with elisidepsin sensitivity in breast cancer cell lines.Protein expression levels of different EMT markers were evaluated by immunocytochemistry (A), western blot (B) and IHC (C). A) Immunocytochemistry of two epithelial (E-cadherin and β-catenin) and four mesenchymal markers (vimentin, Slug, Snail and Twist). Magnification 100x. B) E-cadherin, β-catenin, Slug, Snail, Twist, vimentin and β-actin (loading control) were detected by western blot analysis using 50 µg of total protein. C) Basal levels of E-cadherin, β-catenin and vimentin were analyzed by IHC. Magnification 20x. Each experiment was performed at least in duplicate.
Mentions: We aimed to determine whether the various elisidepsin-sensitive cancer cell lines shared similar basal levels of EMT genes. In the breast cancer cell lines we found E-cadherin expression in the sensitive cell lines. All cell lines had detectable expression of β-catenin, whereas Slug expression was variable and not related to their sensitivity to elisidepsin. Furthermore, Snail expression was only found in MDA-MB-435, and all the cell lines that exhibited levels of Twist-1 and vimentin were less sensitive to the drug (Figs. 2A–C). In contrast, elisidepsin-sensitive pancreatic carcinoma cell lines expressed E-cadherin and β-catenin, whereas the less sensitive cells expressed Slug. Lastly, Snail, Twist-1 and vimentin expression was found in sensitive and insensitive cell lines alike (Figs. 3A–C). To summarize, E-cadherin protein was significantly expressed in the sensitive cell lines independently of their tumoral origin (Mann-Whitney test: p  = 0.0364; Fig. S2), and vimentin was significantly expressed in the less sensitive ones (Mann-Whitney test: p  = 0.0364). On the other hand, Twist-1 and Snail proteins were found in all less sensitive cell lines (Mann Whitney test: p  = 0.0636 and p  = 0.1000, respectively), with the exception of two sensitive cell lines that were positive for vimentin expression (CFPAC and AsPC-1), one sensitive cell line that was positive for Twist-1 expression (CFPAC) and another one that was positive for Snail expression (SKBR3).

Bottom Line: Interestingly, we observed that the basal protein expression levels of EMT markers show a significant correlation with cell viability in response to elisidepsin treatment in a panel of 12 different breast and pancreatic cancer cell lines.In addition, we generated three elisidepsin treatment-resistant cell lines (MCF-7, HPAC and AsPC-1) and analyzed the pattern of expression of different EMT markers in these cells, confirming that acquired resistance to elisidepsin is associated with a switch to the EMT state.These results show that HER3 expression is an important marker of sensitivity to elisidepsin treatment.

View Article: PubMed Central - PubMed

Affiliation: Molecular Pathology Group, Vall d'Hebron Research Institute, Universidad Autonoma of Barcelona, Barcelona, Spain.

ABSTRACT
Elisidepsin (elisidepsin trifluoroacetate, Irvalec®, PM02734) is a new synthetic depsipeptide, a result of the PharmaMar Development Program that seeks synthetic products of marine origin-derived compounds. Elisidepsin is a drug with antiproliferative activity in a wide range of tumors. In the present work we studied and characterized the mechanisms associated with sensitivity and resistance to elisidepsin treatment in a broad panel of tumor cell lines from breast and pancreas carcinomas, focusing on different factors involved in epithelial-mesenchymal transition (EMT) and the use of HER family receptors in predicting the in vitro drug response. Interestingly, we observed that the basal protein expression levels of EMT markers show a significant correlation with cell viability in response to elisidepsin treatment in a panel of 12 different breast and pancreatic cancer cell lines. In addition, we generated three elisidepsin treatment-resistant cell lines (MCF-7, HPAC and AsPC-1) and analyzed the pattern of expression of different EMT markers in these cells, confirming that acquired resistance to elisidepsin is associated with a switch to the EMT state. Furthermore, a direct correlation between basal HER3 expression and sensitivity to elisidepsin was observed; moreover, modulation of HER3 expression levels in different cancer cell lines alter their sensitivities to the drug, making them more resistant when HER3 expression is downregulated by a HER3-specific short hairpin RNA and more sensitive when the receptor is overexpressed. These results show that HER3 expression is an important marker of sensitivity to elisidepsin treatment.

Show MeSH
Related in: MedlinePlus