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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.

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PolyQ-huntingtin promotes mammary cancer cell motility, resistance to cell death and distal metastases in the lungRandom migration assays of PyVT/HdhQ7/Q7 (HdhQ7/Q7) or PyVT/HdhQ111/Q111 (HdhQ111/Q111) cells (3 independent primary cultures; at least 100 cells recorded). ***p-value < 0.0001.Boyden chambers assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least four independent primary cultures in duplicate per genotype). ***p-value < 0.0001.Boyden chambers with matrigel invasion assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least seven independent primary cultures in duplicate per genotype). Representative micrographs of invasion filter membranes after violet staining (t = 48 h) are shown. ***p-value < 0.0001.PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells were cultured in suspension, stained for annexin V and propidium iodide, and analysed by a flow cytometry analysis (four independent experiment, at least four independent primary cultures per genotype). Live, apoptotic and dead cell populations are quantified (live cells, p-value = 0.0099; apoptotic cells, p-value = 0.0835; dead cells, p-value = 0.0169).Lungs from 12 weeks MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 mice are immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (n = 5 lungs per genotype; **p-value = 0.0060).Lungs from mice grafted with MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (MMTV-PyVT/HdhQ7/Q7: n = 5 lungs; MMTV-PyVT/HdhQ111/Q111; n = 4 lungs; *p-value = 0.0266). A second experiment gave similar results.
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fig04: PolyQ-huntingtin promotes mammary cancer cell motility, resistance to cell death and distal metastases in the lungRandom migration assays of PyVT/HdhQ7/Q7 (HdhQ7/Q7) or PyVT/HdhQ111/Q111 (HdhQ111/Q111) cells (3 independent primary cultures; at least 100 cells recorded). ***p-value < 0.0001.Boyden chambers assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least four independent primary cultures in duplicate per genotype). ***p-value < 0.0001.Boyden chambers with matrigel invasion assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least seven independent primary cultures in duplicate per genotype). Representative micrographs of invasion filter membranes after violet staining (t = 48 h) are shown. ***p-value < 0.0001.PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells were cultured in suspension, stained for annexin V and propidium iodide, and analysed by a flow cytometry analysis (four independent experiment, at least four independent primary cultures per genotype). Live, apoptotic and dead cell populations are quantified (live cells, p-value = 0.0099; apoptotic cells, p-value = 0.0835; dead cells, p-value = 0.0169).Lungs from 12 weeks MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 mice are immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (n = 5 lungs per genotype; **p-value = 0.0060).Lungs from mice grafted with MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (MMTV-PyVT/HdhQ7/Q7: n = 5 lungs; MMTV-PyVT/HdhQ111/Q111; n = 4 lungs; *p-value = 0.0266). A second experiment gave similar results.

Mentions: We then tested whether mutant huntingtin played a role in cell motility, which is a functional marker of EMT. We performed random cell migration assays with PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 primary tumour cells (Fig 4A). Cells expressing polyQ-huntingtin moved faster than the corresponding control cells. We also assessed the directed cell migration capacity of the two cell types using Boyden chamber assays, in which PyVT/HdhQ111/Q111 cells transmigrated faster in response to serum than the PyVT/HdhQ7/Q7 cells (Fig 4B). To compare the invasiveness of PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells, we used Boyden chambers containing a layer of ECM proteins on top of the membrane (Fig 4C) and found that PyVT/HdhQ111/Q111 cells were more invasive than PyVT/HdhQ7/Q7 cells. Finally, we assessed cell viability in suspension cultures. PyVT/HdhQ111/Q111 suspension cultures contained more live cells than PyVT/HdhQ7/Q7 suspension cultures as measured by annexin V and propidium iodide (PI) staining and flow cytometric analysis (Fig 4D). In contrast, the percentage of apoptotic and dead cells were lower in the cells that expressed polyQ-huntingtin. Taken together these results show that polyQ-huntingtin expression in cancer cells is associated with enhanced migratory and invasive behaviours, and an elevated resistance to anoikis.


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 promotes mammary cancer cell motility, resistance to cell death and distal metastases in the lungRandom migration assays of PyVT/HdhQ7/Q7 (HdhQ7/Q7) or PyVT/HdhQ111/Q111 (HdhQ111/Q111) cells (3 independent primary cultures; at least 100 cells recorded). ***p-value < 0.0001.Boyden chambers assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least four independent primary cultures in duplicate per genotype). ***p-value < 0.0001.Boyden chambers with matrigel invasion assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least seven independent primary cultures in duplicate per genotype). Representative micrographs of invasion filter membranes after violet staining (t = 48 h) are shown. ***p-value < 0.0001.PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells were cultured in suspension, stained for annexin V and propidium iodide, and analysed by a flow cytometry analysis (four independent experiment, at least four independent primary cultures per genotype). Live, apoptotic and dead cell populations are quantified (live cells, p-value = 0.0099; apoptotic cells, p-value = 0.0835; dead cells, p-value = 0.0169).Lungs from 12 weeks MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 mice are immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (n = 5 lungs per genotype; **p-value = 0.0060).Lungs from mice grafted with MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (MMTV-PyVT/HdhQ7/Q7: n = 5 lungs; MMTV-PyVT/HdhQ111/Q111; n = 4 lungs; *p-value = 0.0266). A second experiment gave similar results.
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fig04: PolyQ-huntingtin promotes mammary cancer cell motility, resistance to cell death and distal metastases in the lungRandom migration assays of PyVT/HdhQ7/Q7 (HdhQ7/Q7) or PyVT/HdhQ111/Q111 (HdhQ111/Q111) cells (3 independent primary cultures; at least 100 cells recorded). ***p-value < 0.0001.Boyden chambers assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least four independent primary cultures in duplicate per genotype). ***p-value < 0.0001.Boyden chambers with matrigel invasion assays for PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells (at least seven independent primary cultures in duplicate per genotype). Representative micrographs of invasion filter membranes after violet staining (t = 48 h) are shown. ***p-value < 0.0001.PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells were cultured in suspension, stained for annexin V and propidium iodide, and analysed by a flow cytometry analysis (four independent experiment, at least four independent primary cultures per genotype). Live, apoptotic and dead cell populations are quantified (live cells, p-value = 0.0099; apoptotic cells, p-value = 0.0835; dead cells, p-value = 0.0169).Lungs from 12 weeks MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 mice are immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (n = 5 lungs per genotype; **p-value = 0.0060).Lungs from mice grafted with MMTV-PyVT/HdhQ7/Q7 and MMTV-PyVT/HdhQ111/Q111 tumours immunostained with anti-PyVT antibodies. The mean number of metastatic lesions per section is shown (MMTV-PyVT/HdhQ7/Q7: n = 5 lungs; MMTV-PyVT/HdhQ111/Q111; n = 4 lungs; *p-value = 0.0266). A second experiment gave similar results.
Mentions: We then tested whether mutant huntingtin played a role in cell motility, which is a functional marker of EMT. We performed random cell migration assays with PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 primary tumour cells (Fig 4A). Cells expressing polyQ-huntingtin moved faster than the corresponding control cells. We also assessed the directed cell migration capacity of the two cell types using Boyden chamber assays, in which PyVT/HdhQ111/Q111 cells transmigrated faster in response to serum than the PyVT/HdhQ7/Q7 cells (Fig 4B). To compare the invasiveness of PyVT/HdhQ7/Q7 and PyVT/HdhQ111/Q111 cells, we used Boyden chambers containing a layer of ECM proteins on top of the membrane (Fig 4C) and found that PyVT/HdhQ111/Q111 cells were more invasive than PyVT/HdhQ7/Q7 cells. Finally, we assessed cell viability in suspension cultures. PyVT/HdhQ111/Q111 suspension cultures contained more live cells than PyVT/HdhQ7/Q7 suspension cultures as measured by annexin V and propidium iodide (PI) staining and flow cytometric analysis (Fig 4D). In contrast, the percentage of apoptotic and dead cells were lower in the cells that expressed polyQ-huntingtin. Taken together these results show that polyQ-huntingtin expression in cancer cells is associated with enhanced migratory and invasive behaviours, and an elevated resistance to anoikis.

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