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Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis

An Hendrix UGent, Dawn Maynard, Patrick Pauwels UGent, Geert Braems UGent, Hannelore Denys UGent, Rudy Van den Broecke UGent, Jo Lambert UGent, Simon Van Belle UGent, Veronique Cocquyt UGent and Christian Gespach, et al. (2010) JOURNAL OF THE NATIONAL CANCER INSTITUTE. 102(12). p.866-880
abstract
Background Secretory GTPases like Rab27B control vesicle exocytosis and deliver critical proinvasive growth regulators into the tumor microenvironment. The expression and role of Rab27B in breast cancer were unknown. Methods Expression of green fluorescent protein (GFP) fused with wild-type Rab3D, Rab27A, or Rab27B, or Rab27B point mutants defective in GTP/GDP binding or geranylgeranylation, or transient silencing RNA to the same proteins was used to study Rab27B in estrogen receptor (ER)-positive human breast cancer cell lines (MCF-7, T47D, and ZR75.1). Cell cycle progression was evaluated by flow cytometry, western blotting, and measurement of cell proliferation rates, and invasion was assessed using Matrigel and native type I collagen substrates. Orthotopic tumor growth, local invasion, and metastasis were analyzed in mouse xenograft models. Mass spectrometry identified proinvasive growth regulators that were secreted in the presence of Rab27B. Rab27B protein levels were evaluated by immunohistochemistry in 59 clinical breast cancer specimens, and Rab3D, Rab27A, and Rab27B mRNA levels were analyzed by quantitative real-time polymerase chain reaction in 20 specimens. Statistical tests were two-sided. Results Increased expression of Rab27B promoted G(1) to S phase cell cycle transition, proliferation and invasiveness of cells in culture, and invasive tumor growth and hemorrhagic ascites production in a xenograft mouse model (n = 10; at 10 weeks, survival of MCF-7 GFP- vs GFP-Rab27B-injected mice was 100% vs 62.5%, hazard ratio = 0.26, 95% confidence interval = 0.08 to 0.88, P = .03). Mass spectrometric analysis of purified Rab27B-secretory vesicles identified heat-shock protein 90alpha as key proinvasive growth regulator. Heat-shock protein 90alpha secretion was Rab27B-dependent and was required for matrix metalloproteinase-2 activation. All Rab27B-mediated functional responses were GTP- and geranylgeranyl-dependent. Presence of endogenous Rab27B mRNA and protein, but not of Rab3D or Rab27A mRNA, was associated with lymph node metastasis (P < .001) and differentiation grade (P = .001) in ER-positive human breast tumors. Conclusions Rab27B regulates invasive growth and metastasis in ER-positive breast cancer cell lines, and increased expression is associated with poor prognosis in humans.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
MASS-SPECTROMETRY, MATRIX METALLOPROTEINASES, CELL-MIGRATION, EXOCYTOSIS, GRANULES, HSP90, Rab GTPases, vesicle transport, estrogen receptor, Breast cancer
journal title
JOURNAL OF THE NATIONAL CANCER INSTITUTE
J. Natl. Cancer Inst.
volume
102
issue
12
pages
866 - 880
Web of Science type
Article
Web of Science id
000279925200008
JCR category
ONCOLOGY
JCR impact factor
14.697 (2010)
JCR rank
6/181 (2010)
JCR quartile
1 (2010)
ISSN
0027-8874
DOI
10.1093/jnci/djq153
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
968177
handle
http://hdl.handle.net/1854/LU-968177
date created
2010-06-02 13:45:27
date last changed
2010-10-05 11:51:31
@article{968177,
  abstract     = {Background Secretory GTPases like Rab27B control vesicle exocytosis and deliver critical proinvasive growth regulators into the tumor microenvironment. The expression and role of Rab27B in breast cancer were unknown. Methods Expression of green fluorescent protein (GFP) fused with wild-type Rab3D, Rab27A, or Rab27B, or Rab27B point mutants defective in GTP/GDP binding or geranylgeranylation, or transient silencing RNA to the same proteins was used to study Rab27B in estrogen receptor (ER)-positive human breast cancer cell lines (MCF-7, T47D, and ZR75.1). Cell cycle progression was evaluated by flow cytometry, western blotting, and measurement of cell proliferation rates, and invasion was assessed using Matrigel and native type I collagen substrates. Orthotopic tumor growth, local invasion, and metastasis were analyzed in mouse xenograft models. Mass spectrometry identified proinvasive growth regulators that were secreted in the presence of Rab27B. Rab27B protein levels were evaluated by immunohistochemistry in 59 clinical breast cancer specimens, and Rab3D, Rab27A, and Rab27B mRNA levels were analyzed by quantitative real-time polymerase chain reaction in 20 specimens. Statistical tests were two-sided. Results Increased expression of Rab27B promoted G(1) to S phase cell cycle transition, proliferation and invasiveness of cells in culture, and invasive tumor growth and hemorrhagic ascites production in a xenograft mouse model (n = 10; at 10 weeks, survival of MCF-7 GFP- vs GFP-Rab27B-injected mice was 100\% vs 62.5\%, hazard ratio = 0.26, 95\% confidence interval = 0.08 to 0.88, P = .03). Mass spectrometric analysis of purified Rab27B-secretory vesicles identified heat-shock protein 90alpha as key proinvasive growth regulator. Heat-shock protein 90alpha secretion was Rab27B-dependent and was required for matrix metalloproteinase-2 activation. All Rab27B-mediated functional responses were GTP- and geranylgeranyl-dependent. Presence of endogenous Rab27B mRNA and protein, but not of Rab3D or Rab27A mRNA, was associated with lymph node metastasis (P {\textlangle} .001) and differentiation grade (P = .001) in ER-positive human breast tumors. Conclusions Rab27B regulates invasive growth and metastasis in ER-positive breast cancer cell lines, and increased expression is associated with poor prognosis in humans.},
  author       = {Hendrix, An and Maynard, Dawn and Pauwels, Patrick and Braems, Geert and Denys, Hannelore and Van den Broecke, Rudy and Lambert, Jo and Van Belle, Simon and Cocquyt, Veronique and Gespach, Christian and Bracke, Marc and Seabra, Miguel C and Gahl, William A and De Wever, Olivier and Westbroek, Wendy},
  issn         = {0027-8874},
  journal      = {JOURNAL OF THE NATIONAL CANCER INSTITUTE},
  keyword      = {MASS-SPECTROMETRY,MATRIX METALLOPROTEINASES,CELL-MIGRATION,EXOCYTOSIS,GRANULES,HSP90,Rab GTPases,vesicle transport,estrogen receptor,Breast cancer},
  language     = {eng},
  number       = {12},
  pages        = {866--880},
  title        = {Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis},
  url          = {http://dx.doi.org/10.1093/jnci/djq153},
  volume       = {102},
  year         = {2010},
}

Chicago
Hendrix, An, Dawn Maynard, Patrick Pauwels, Geert Braems, Hannelore Denys, Rudy Van den Broecke, Jo Lambert, et al. 2010. “Effect of the Secretory Small GTPase Rab27B on Breast Cancer Growth, Invasion, and Metastasis.” Journal of the National Cancer Institute 102 (12): 866–880.
APA
Hendrix, A., Maynard, D., Pauwels, P., Braems, G., Denys, H., Van den Broecke, R., Lambert, J., et al. (2010). Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis. JOURNAL OF THE NATIONAL CANCER INSTITUTE, 102(12), 866–880.
Vancouver
1.
Hendrix A, Maynard D, Pauwels P, Braems G, Denys H, Van den Broecke R, et al. Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis. JOURNAL OF THE NATIONAL CANCER INSTITUTE. 2010;102(12):866–80.
MLA
Hendrix, An, Dawn Maynard, Patrick Pauwels, et al. “Effect of the Secretory Small GTPase Rab27B on Breast Cancer Growth, Invasion, and Metastasis.” JOURNAL OF THE NATIONAL CANCER INSTITUTE 102.12 (2010): 866–880. Print.