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Oncogene-like induction of cellular invasion from centrosome amplification.

Godinho SA, Picone R, Burute M, Dagher R, Su Y, Leung CT, Polyak K, Brugge JS, Théry M, Pellman D - Nature (2014)

Bottom Line: Centrosome amplification is poorly tolerated by non-transformed cells and, in the absence of selection, extra centrosomes are spontaneously lost.Our data indicate that, through increased centrosomal microtubule nucleation, centrosome amplification increases Rac1 activity, which disrupts normal cell-cell adhesion and promotes invasion.These findings demonstrate that centrosome amplification, a structural alteration of the cytoskeleton, can promote features of malignant transformation.

View Article: PubMed Central - PubMed

Affiliation: 1] Howard Hughes Medical Institute, Department of Pediatric Oncology, Dana-Farber Cancer Institute and Pediatric Hematology/Oncology, Children's Hospital, Boston, Massachusetts 02115, USA [2] Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA [3] Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK (S.A.G.); Department of Pharmacology, University of Minnesota, Minneapolis, Minnesota 55455, USA (C.T.L.).

ABSTRACT
Centrosome amplification has long been recognized as a feature of human tumours; however, its role in tumorigenesis remains unclear. Centrosome amplification is poorly tolerated by non-transformed cells and, in the absence of selection, extra centrosomes are spontaneously lost. Thus, the high frequency of centrosome amplification, particularly in more aggressive tumours, raises the possibility that extra centrosomes could, in some contexts, confer advantageous characteristics that promote tumour progression. Using a three-dimensional model system and other approaches to culture human mammary epithelial cells, we find that centrosome amplification triggers cell invasion. This invasive behaviour is similar to that induced by overexpression of the breast cancer oncogene ERBB2 (ref. 4) and indeed enhances invasiveness triggered by ERBB2. Our data indicate that, through increased centrosomal microtubule nucleation, centrosome amplification increases Rac1 activity, which disrupts normal cell-cell adhesion and promotes invasion. These findings demonstrate that centrosome amplification, a structural alteration of the cytoskeleton, can promote features of malignant transformation.

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The induction of aneuploidy does not generate invasive acinia, Western blot showing depletion of MCAK by siRNA (48hrs). *Marks a non-specific band. b, Quantification of chromosome number, shown as the percentage deviation from the mode, in cells before adding to 3-D cultures (48hrs) and after 3-D cultures (~100 spreads were scored per condition). Significance was determined by Chi-squared testing to compare the frequencies of nominal variables (*, p<0.05). c, Fraction of cells with centrosome amplification. Error bars represent mean ± SE from 3 independent experiments. d, Fraction of invasive acini. Error bars represent mean ± SE from 3 independent experiments. p-value derived from unpaired two-tailed t-test (***, p<0.0005; **, p<0.005; n.s. not significant). e, SNP analysis of cells with extra centrosomes and depleted of MCAK. Shown are log2 copy number ratios of the indicated samples relative to their controls.
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Figure 2: The induction of aneuploidy does not generate invasive acinia, Western blot showing depletion of MCAK by siRNA (48hrs). *Marks a non-specific band. b, Quantification of chromosome number, shown as the percentage deviation from the mode, in cells before adding to 3-D cultures (48hrs) and after 3-D cultures (~100 spreads were scored per condition). Significance was determined by Chi-squared testing to compare the frequencies of nominal variables (*, p<0.05). c, Fraction of cells with centrosome amplification. Error bars represent mean ± SE from 3 independent experiments. d, Fraction of invasive acini. Error bars represent mean ± SE from 3 independent experiments. p-value derived from unpaired two-tailed t-test (***, p<0.0005; **, p<0.005; n.s. not significant). e, SNP analysis of cells with extra centrosomes and depleted of MCAK. Shown are log2 copy number ratios of the indicated samples relative to their controls.

Mentions: Author contributions S.A.G and D.P. designed the experiments and wrote the manuscript. S.A.G conceived, conducted and performed data analysis for most experiments. R.P. conceived and conducted all FRET experiments and Fig. 3a. M.T and M.B. contributed with micro-pattern fabrication, Fig. 3d, Extended Data Fig. 6e-d and Extended Data Fig. 8c-e. R.D. contributed with Fig. 2b and Extended Data Fig. 5a. Y.S., K.P., C.L. and J.B. provided assistance with 3-dimentional cultures. All authors contributed with discussions and edited the manuscript.


Oncogene-like induction of cellular invasion from centrosome amplification.

Godinho SA, Picone R, Burute M, Dagher R, Su Y, Leung CT, Polyak K, Brugge JS, Théry M, Pellman D - Nature (2014)

The induction of aneuploidy does not generate invasive acinia, Western blot showing depletion of MCAK by siRNA (48hrs). *Marks a non-specific band. b, Quantification of chromosome number, shown as the percentage deviation from the mode, in cells before adding to 3-D cultures (48hrs) and after 3-D cultures (~100 spreads were scored per condition). Significance was determined by Chi-squared testing to compare the frequencies of nominal variables (*, p<0.05). c, Fraction of cells with centrosome amplification. Error bars represent mean ± SE from 3 independent experiments. d, Fraction of invasive acini. Error bars represent mean ± SE from 3 independent experiments. p-value derived from unpaired two-tailed t-test (***, p<0.0005; **, p<0.005; n.s. not significant). e, SNP analysis of cells with extra centrosomes and depleted of MCAK. Shown are log2 copy number ratios of the indicated samples relative to their controls.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4061398&req=5

Figure 2: The induction of aneuploidy does not generate invasive acinia, Western blot showing depletion of MCAK by siRNA (48hrs). *Marks a non-specific band. b, Quantification of chromosome number, shown as the percentage deviation from the mode, in cells before adding to 3-D cultures (48hrs) and after 3-D cultures (~100 spreads were scored per condition). Significance was determined by Chi-squared testing to compare the frequencies of nominal variables (*, p<0.05). c, Fraction of cells with centrosome amplification. Error bars represent mean ± SE from 3 independent experiments. d, Fraction of invasive acini. Error bars represent mean ± SE from 3 independent experiments. p-value derived from unpaired two-tailed t-test (***, p<0.0005; **, p<0.005; n.s. not significant). e, SNP analysis of cells with extra centrosomes and depleted of MCAK. Shown are log2 copy number ratios of the indicated samples relative to their controls.
Mentions: Author contributions S.A.G and D.P. designed the experiments and wrote the manuscript. S.A.G conceived, conducted and performed data analysis for most experiments. R.P. conceived and conducted all FRET experiments and Fig. 3a. M.T and M.B. contributed with micro-pattern fabrication, Fig. 3d, Extended Data Fig. 6e-d and Extended Data Fig. 8c-e. R.D. contributed with Fig. 2b and Extended Data Fig. 5a. Y.S., K.P., C.L. and J.B. provided assistance with 3-dimentional cultures. All authors contributed with discussions and edited the manuscript.

Bottom Line: Centrosome amplification is poorly tolerated by non-transformed cells and, in the absence of selection, extra centrosomes are spontaneously lost.Our data indicate that, through increased centrosomal microtubule nucleation, centrosome amplification increases Rac1 activity, which disrupts normal cell-cell adhesion and promotes invasion.These findings demonstrate that centrosome amplification, a structural alteration of the cytoskeleton, can promote features of malignant transformation.

View Article: PubMed Central - PubMed

Affiliation: 1] Howard Hughes Medical Institute, Department of Pediatric Oncology, Dana-Farber Cancer Institute and Pediatric Hematology/Oncology, Children's Hospital, Boston, Massachusetts 02115, USA [2] Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA [3] Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK (S.A.G.); Department of Pharmacology, University of Minnesota, Minneapolis, Minnesota 55455, USA (C.T.L.).

ABSTRACT
Centrosome amplification has long been recognized as a feature of human tumours; however, its role in tumorigenesis remains unclear. Centrosome amplification is poorly tolerated by non-transformed cells and, in the absence of selection, extra centrosomes are spontaneously lost. Thus, the high frequency of centrosome amplification, particularly in more aggressive tumours, raises the possibility that extra centrosomes could, in some contexts, confer advantageous characteristics that promote tumour progression. Using a three-dimensional model system and other approaches to culture human mammary epithelial cells, we find that centrosome amplification triggers cell invasion. This invasive behaviour is similar to that induced by overexpression of the breast cancer oncogene ERBB2 (ref. 4) and indeed enhances invasiveness triggered by ERBB2. Our data indicate that, through increased centrosomal microtubule nucleation, centrosome amplification increases Rac1 activity, which disrupts normal cell-cell adhesion and promotes invasion. These findings demonstrate that centrosome amplification, a structural alteration of the cytoskeleton, can promote features of malignant transformation.

Show MeSH
Related in: MedlinePlus