Limits...
The Huntington disease protein accelerates breast tumour development and metastasis through ErbB2/HER2 signalling.

Moreira Sousa C, McGuire JR, Thion MS, Gentien D, de la Grange P, Tezenas du Montcel S, Vincent-Salomon A, Durr A, Humbert S - EMBO Mol Med (2013)

Bottom Line: In agreement, mutant huntingtin accelerates epithelial to mesenchymal transition and enhances cell motility and invasion.Finally, we report that in HD, the dynamin dependent endocytosis of the ErbB2/HER2 receptor tyrosine kinase is reduced.This leads to its accumulation and to subsequent increases in cell motility and proliferation.

View Article: PubMed Central - PubMed

Affiliation: Institut Curie, Paris, France; CNRS UMR 3306, Orsay, France.

Show MeSH

Related in: MedlinePlus

PolyQ-huntingtin accelerates EMTImmunostaining of sections from MMTV-PyVT/HdhQ7/Q7 (HdhQ7/Q7) and MMTV-PyVT/HdhQ111/Q111 (HdhQ111/Q111) tumours (originating from the 3rd mammary gland, 14 weeks) for endogenous E-cadherin, β-catenin and α-SMA.Immunoblotting of total extracts from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours (originating from the 3rd mammary gland, 14 weeks, n = 6 tumours per genotype, two independent immunoblotting) for the presence of ZO-1, E-cadherin, β-catenin, vimentin and α-tubulin. ZO-1: ***p-value = 0.0007; E-cadherin: *p-value = 0.0295; β-catenin: *p-value = 0.0238; vimentin: **p-value = 0.0039.Differential interfering contrast images of cells from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 dissociated tumours.Immunoblotting of extracts from PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells as in (B).PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells are immunostained for endogenous E-cadherin and β-catenin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3569645&req=5

fig03: PolyQ-huntingtin accelerates EMTImmunostaining of sections from MMTV-PyVT/HdhQ7/Q7 (HdhQ7/Q7) and MMTV-PyVT/HdhQ111/Q111 (HdhQ111/Q111) tumours (originating from the 3rd mammary gland, 14 weeks) for endogenous E-cadherin, β-catenin and α-SMA.Immunoblotting of total extracts from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours (originating from the 3rd mammary gland, 14 weeks, n = 6 tumours per genotype, two independent immunoblotting) for the presence of ZO-1, E-cadherin, β-catenin, vimentin and α-tubulin. ZO-1: ***p-value = 0.0007; E-cadherin: *p-value = 0.0295; β-catenin: *p-value = 0.0238; vimentin: **p-value = 0.0039.Differential interfering contrast images of cells from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 dissociated tumours.Immunoblotting of extracts from PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells as in (B).PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells are immunostained for endogenous E-cadherin and β-catenin.

Mentions: To test this hypothesis, we analysed primary tumour sections by immunohistochemistry (Fig 3A). Lowered levels of the cell–cell adhesion proteins E-cadherin and β-catenin were observed in the MMTV-PyVT/HdhQ111/Q111 tumours, while the mesenchymal marker α-smooth muscle actin (α-SMA) was increased. We then analysed extracts from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours by immunoblotting (Fig 3B). The MMTV-PyVT/HdhQ111/Q111 tumours had lower levels of the tight junction protein zonula occludens 1 (ZO1), E-cadherin and β-catenin, and an increased level of the mesenchymal marker vimentin compared to MMTV-PyVT/HdhQ7/Q7 tumours. Similarly, E-cadherin, β-catenin and vimentin levels were affected in MMTV-ErbB2/HdhQ111/Q111 tumours as compared to MMTV-ErbB2/HdhQ7/Q7 tumours (Supporting Information Fig S2C). We next derived primary tumour cells from the MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours (PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111, respectively). Doubling-time measurements showed no significant difference between wild-type (PyVT/HdhQ7/Q7) and polyQ-huntingtin-expressing cells (PyVT/HdhQ111/Q111) (12.38 ± 0.25 h and 12.65 ± 0.22 h, respectively, PLSD Fisher test p-value = 0.4357). However, further confirming the microarray data, polyQ-huntingtin expression led to a mesenchymal-like phenotype in the PyVT/HdhQ111/Q111 cells, which became scattered and elongated compared to phenotype of the PyVT/HdhQ7/Q7 cells (Fig 3C). Also, the observations done on tumours with respect to the levels of ZO1, E-cadherin, β-catenin and vimentin were confirmed in an immunoblot analysis of extracts from PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 tumour cells (Fig 3D). Immunostaining confirmed that E-cadherin and β-catenin production were substantially lower in polyQ-huntingtin-expressing cells (Fig 3E). Furthermore, β-catenin exhibited membrane localization in PyVT/HdhQ7/Q7 cells and diffuse cytoplasmic localization in PyVT/HdhQ111/Q111 cells, which is consistent with decreased cellular adhesion. In summary, mutant huntingtin expression in primary tumour tissue and in tumour-derived cells affects the levels of known cell adhesion markers and mesenchymal markers. Furthermore, when mutant huntingtin is expressed, tumour cells in culture adopt an altered morphology resembling a mesenchymal phenotype.


The Huntington disease protein accelerates breast tumour development and metastasis through ErbB2/HER2 signalling.

Moreira Sousa C, McGuire JR, Thion MS, Gentien D, de la Grange P, Tezenas du Montcel S, Vincent-Salomon A, Durr A, Humbert S - EMBO Mol Med (2013)

PolyQ-huntingtin accelerates EMTImmunostaining of sections from MMTV-PyVT/HdhQ7/Q7 (HdhQ7/Q7) and MMTV-PyVT/HdhQ111/Q111 (HdhQ111/Q111) tumours (originating from the 3rd mammary gland, 14 weeks) for endogenous E-cadherin, β-catenin and α-SMA.Immunoblotting of total extracts from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours (originating from the 3rd mammary gland, 14 weeks, n = 6 tumours per genotype, two independent immunoblotting) for the presence of ZO-1, E-cadherin, β-catenin, vimentin and α-tubulin. ZO-1: ***p-value = 0.0007; E-cadherin: *p-value = 0.0295; β-catenin: *p-value = 0.0238; vimentin: **p-value = 0.0039.Differential interfering contrast images of cells from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 dissociated tumours.Immunoblotting of extracts from PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells as in (B).PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells are immunostained for endogenous E-cadherin and β-catenin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3569645&req=5

fig03: PolyQ-huntingtin accelerates EMTImmunostaining of sections from MMTV-PyVT/HdhQ7/Q7 (HdhQ7/Q7) and MMTV-PyVT/HdhQ111/Q111 (HdhQ111/Q111) tumours (originating from the 3rd mammary gland, 14 weeks) for endogenous E-cadherin, β-catenin and α-SMA.Immunoblotting of total extracts from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours (originating from the 3rd mammary gland, 14 weeks, n = 6 tumours per genotype, two independent immunoblotting) for the presence of ZO-1, E-cadherin, β-catenin, vimentin and α-tubulin. ZO-1: ***p-value = 0.0007; E-cadherin: *p-value = 0.0295; β-catenin: *p-value = 0.0238; vimentin: **p-value = 0.0039.Differential interfering contrast images of cells from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 dissociated tumours.Immunoblotting of extracts from PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells as in (B).PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells are immunostained for endogenous E-cadherin and β-catenin.
Mentions: To test this hypothesis, we analysed primary tumour sections by immunohistochemistry (Fig 3A). Lowered levels of the cell–cell adhesion proteins E-cadherin and β-catenin were observed in the MMTV-PyVT/HdhQ111/Q111 tumours, while the mesenchymal marker α-smooth muscle actin (α-SMA) was increased. We then analysed extracts from MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours by immunoblotting (Fig 3B). The MMTV-PyVT/HdhQ111/Q111 tumours had lower levels of the tight junction protein zonula occludens 1 (ZO1), E-cadherin and β-catenin, and an increased level of the mesenchymal marker vimentin compared to MMTV-PyVT/HdhQ7/Q7 tumours. Similarly, E-cadherin, β-catenin and vimentin levels were affected in MMTV-ErbB2/HdhQ111/Q111 tumours as compared to MMTV-ErbB2/HdhQ7/Q7 tumours (Supporting Information Fig S2C). We next derived primary tumour cells from the MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours (PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111, respectively). Doubling-time measurements showed no significant difference between wild-type (PyVT/HdhQ7/Q7) and polyQ-huntingtin-expressing cells (PyVT/HdhQ111/Q111) (12.38 ± 0.25 h and 12.65 ± 0.22 h, respectively, PLSD Fisher test p-value = 0.4357). However, further confirming the microarray data, polyQ-huntingtin expression led to a mesenchymal-like phenotype in the PyVT/HdhQ111/Q111 cells, which became scattered and elongated compared to phenotype of the PyVT/HdhQ7/Q7 cells (Fig 3C). Also, the observations done on tumours with respect to the levels of ZO1, E-cadherin, β-catenin and vimentin were confirmed in an immunoblot analysis of extracts from PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 tumour cells (Fig 3D). Immunostaining confirmed that E-cadherin and β-catenin production were substantially lower in polyQ-huntingtin-expressing cells (Fig 3E). Furthermore, β-catenin exhibited membrane localization in PyVT/HdhQ7/Q7 cells and diffuse cytoplasmic localization in PyVT/HdhQ111/Q111 cells, which is consistent with decreased cellular adhesion. In summary, mutant huntingtin expression in primary tumour tissue and in tumour-derived cells affects the levels of known cell adhesion markers and mesenchymal markers. Furthermore, when mutant huntingtin is expressed, tumour cells in culture adopt an altered morphology resembling a mesenchymal phenotype.

Bottom Line: In agreement, mutant huntingtin accelerates epithelial to mesenchymal transition and enhances cell motility and invasion.Finally, we report that in HD, the dynamin dependent endocytosis of the ErbB2/HER2 receptor tyrosine kinase is reduced.This leads to its accumulation and to subsequent increases in cell motility and proliferation.

View Article: PubMed Central - PubMed

Affiliation: Institut Curie, Paris, France; CNRS UMR 3306, Orsay, France.

Show MeSH
Related in: MedlinePlus