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BMP signalling controls the malignant potential of ascites-derived human epithelial ovarian cancer spheroids via AKT kinase activation

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Abstract

Epithelial ovarian cancer (EOC) cells have the ability to form multi-cellular aggregates in malignant ascites which dramatically alters cell signalling, survival, and metastatic potential. Herein, we demonstrate that patient ascites-derived EOC cells down-regulate endogenous bone morphogenetic protein (BMP) signalling by decreasing BMP ligand expression when grown in suspension culture to form spheroids. Enforced BMP signalling in these cells via constitutively-active BMP type I ALK3QD receptor expression causes the formation of smaller, more loosely-aggregated spheroids. Additionally, ALK3QD-expressing spheroids have an increased rate of adhesion and dispersion upon reattachment to substratum. Inhibition of endogenous BMP signalling using recombinant Noggin or small molecule inhibitor LDN-193189, on the other hand, opposed these phenotypic changes. To identify potential targets that impact the phenotype of EOC spheroids due to activated BMP signalling, we performed genome-wide expression analyses using Affymetrix arrays. Using the online Connectivity Map resource, the BMP signalling gene expression signature revealed that the AKT pathway is induced by activated BMP signalling in EOC cells; this finding was further validated by phospho-AKT immuno-blotting. In fact, treatment of EOC spheroids with an AKT inhibitor, Akti-1/2, reduced BMP-stimulated cell dispersion during reattachment as compared to controls. Thus, we have identified AKT as being one important downstream component of activated BMP signalling on EOC spheroid pathobiology, which may have important implications on the metastatic potential of this malignancy.

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Abbreviations

EOC:

Epithelial ovarian cancer

BMP:

Bone morphogenetic protein

TGFβ:

Transforming growth factor beta

OSE:

Ovarian surface epithelium

MIS:

Mullerian inhibiting substance

EMT:

Epithelial-mesenchymal transition

FBS:

Fetal bovine serum

ULA:

Ultra Low-Attachment

GFP:

Green fluorescent protein

HA:

Hemagglutinin

NG:

Noggin

CMAP:

Connectivity Map

PI3K:

Phosphatidylinositol 3-kinase

mTOR:

Mammalian target of rapamycin

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Acknowledgments

We acknowledge the essential contribution of our colleagues the gynaecologic oncology surgeons Monique Bertrand, Akira Sugimoto, and Michel Préfontaine at the London Regional Cancer Program in providing all clinical specimens used in this study, and Christine Gawlik for assistance with retrieval of clinical data. We thank Dominik Dobransky and Sara Fell for assisting with experiments, Carly Shanks for data analysis and graphic artwork, and Dr. Christopher Pin for insightful comments on this manuscript. We are also extremely grateful to the women with ovarian cancer who generously donated their ascites samples to support our research. This research was supported by funding from an Ovarian Cancer Canada—Canadian Institutes for Health Research partnership grant (Grant No. OVA-94083) and the Canadian Cancer Society (Grant No. 20109). T. Peart and R. Correa are supported by graduate scholarships from the Strategic Training Program in Cancer Research and Technology Transfer Program with funds from Canadian Institutes for Health Research.

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Authors and Affiliations

  1. Translational Ovarian Cancer Research Program, London Regional Cancer Program, 790 Commissioners Road East, Room A4-836, London, ON, N6A 4L6, Canada

    Teresa M. Peart, Rohann J. M. Correa, Yudith Ramos Valdes, Gabriel E. DiMattia & Trevor G. Shepherd

  2. Departments of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Teresa M. Peart & Trevor G. Shepherd

  3. Departments of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Rohann J. M. Correa & Gabriel E. DiMattia

  4. Departments of Oncology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Gabriel E. DiMattia & Trevor G. Shepherd

  5. Departments of Obstetrics and Gynaecology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Gabriel E. DiMattia & Trevor G. Shepherd

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  1. Teresa M. Peart
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  2. Rohann J. M. Correa
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  3. Yudith Ramos Valdes
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  4. Gabriel E. DiMattia
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  5. Trevor G. Shepherd
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Correspondence to Trevor G. Shepherd.

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Peart, T.M., Correa, R.J.M., Valdes, Y.R. et al. BMP signalling controls the malignant potential of ascites-derived human epithelial ovarian cancer spheroids via AKT kinase activation. Clin Exp Metastasis 29, 293–313 (2012). https://doi.org/10.1007/s10585-011-9451-3

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  • DOI: https://doi.org/10.1007/s10585-011-9451-3

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