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Osteopontin Knockdown Suppresses Tumorigenicity of Human Metastatic Breast Carcinoma, MDA-MB-435

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Abstract

Elevated expression of osteopontin (OPN), a secreted phosphoglycoprotein, is frequently associated with many transformed cell lines. Various studies suggest that OPN may contribute to tumor progression as well as metastasis in multiple tumor types. High levels of OPN have been reported in patients with metastatic cancers, including breast. We found that the expression of OPN corroborates with the aggressive phenotype of the breast cancer cells i.e. the expression of OPN is acquired as the breast cancer cells become more aggressive. To assess the role(s) of OPN in breast carcinoma, expression of endogenous OPN was knocked down in metastatic MDA-MB-435 human breast carcinoma cells using RNA interference. We targeted multiple regions of the OPN transcript for RNA interference, along with ‘scrambled’ and ‘non-targeting siRNA pool’ controls to distinguish between target-specific and potential off-target effects including interferon-response gene (PeIF2-α) induction. The OPN knockdown by shRNA suppressed tumor take in immunocompromised mice. The ‘silenced’ cells also showed significantly lower invasion and migration in modified Boyden chamber assays and reduced ability to grow in soft agar. Thus, in addition to the widely reported roles of OPN in late stages of tumor progression, these results provide functional evidence that OPN contributes to breast tumor growth as well.

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Abbreviations

CMF-DPBS:

Calcium- and magnesium-free Dulbecco’s phosphate buffered saline

DMEM-F12:

Mixture (1:1) Dulbecco’s-modified minimum essential medium and Ham’s F-12 medium

HBSS:

Hank’s balanced salt solution

SDS:

Sodium dodecyl sulfate

PAGE:

Poly acrylamide gel electrophoresis

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Acknowledgements

This work was supported by grants from The Susan G. Komen Breast Cancer Foundation BCTR0402317 (LAS), U.S. Public Health service, CA87728 (DRW), CA89019 (DRW), U.S. Army Medical Research and Materiel Command DAMD-17-02-0541 (DRW), Ontario Cancer Research Network 04-MAY-00089 (AFC) and the National Foundation for Cancer Research. AFC is Canada Research Chair in Oncology. L. A. Shevde was a recipient of a postdoctoral fellowship from the Susan G. Komen Breast Cancer Foundation (PDF 2000-218) and␣ACS-IRG 60-0010-44 and CA13148-31. L. A. Shevde and R.␣S. Samant were both recipients of pilot project grants from the UAB Breast SPORE (CA89019) and R. S. Samant is currently supported by the Susan G. Komen Breast Cancer Foundation (BCTR0503488).

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Authors and Affiliations

  1. University of South Alabama-Mitchell Cancer Institute, 307 N. University Blvd., Mobile, AL, 36688-0002, USA

    Lalita A. Shevde, Rajeev S. Samant & Brandon J. Metge

  2. Department of Pathology, The University of Alabama at Birmingham, Birmingham, AL, 35294-0019, USA

    Lalita A. Shevde, Rajeev S. Samant, Jason C. Paik, Andra R. Frost & Danny R. Welch

  3. Comprehensive Cancer Center, The University of Alabama at Birmingham, Birmingham, Alabama, USA

    Lalita A. Shevde, Rajeev S. Samant, Andra R. Frost & Danny R. Welch

  4. NFCR-Center for Metastasis Research, University of Alabama, Birmingham, Alabama, USA

    Danny R. Welch

  5. The London Regional Cancer Program, London, Ontario, Canada

    Ann F. Chambers

  6. Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Lerner School of Medicine, Cleveland, Ohio, USA

    Graham Casey

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  1. Lalita A. Shevde
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  2. Rajeev S. Samant
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Correspondence to Lalita A. Shevde.

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Shevde, L.A., Samant, R.S., Paik, J.C. et al. Osteopontin Knockdown Suppresses Tumorigenicity of Human Metastatic Breast Carcinoma, MDA-MB-435. Clin Exp Metastasis 23, 123–133 (2006). https://doi.org/10.1007/s10585-006-9013-2

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