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Parathyroid hormone-related protein and ezrin are up-regulated in human lung cancer bone metastases

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Abstract

Lung cancer often metastasizes to bone in patients with advanced disease. Identification of the factors involved in the interactions between lung cancer cells and bone will improve the prevention and treatment of bone metastases. We identified changes in metastasis-related gene expression of human HARA lung squamous carcinoma cells co-cultured with neonatal mouse calvariae using a pathway-specific microarray analysis. Nine genes were up-regulated and two genes down-regulated in HARA cells co-cultured with mouse calvariae. Five of the nine up-regulated genes, including caveolin 1, CD44, EphB2, ezrin, and Parathyroid hormone-related protein (PTHrP), and one down-regulated gene, SLPI, were further confirmed by Reverse transcription-polymerase chain reaction (RT-PCR). A mouse model was subsequently used to study the role of PTHrP and ezrin in bone metastasis in vivo. PTHrP (all three isoforms) and ezrin were up-regulated in HARA cells at sites of bone metastasis as detected by RT-PCR and immunohistochemistry. The PTHrP 141 mRNA isoform was increased by the greatest extent (13.9-fold) in bone metastases compared to PTHrP 139 and PTHrP 173 mRNA. We then generated a HARA cell line in which PTHrP expression was inducibly silenced by RNA interference. Silencing of PTHrP expression caused significant reduction of submembranous F-actin and decreased HARA cell invasion. Ezrin up-regulation was confirmed by Western blots on HARA cells co-cultured with adult mouse long bones. Further, Transforming growth factor beta (TGF-β) was identified as one of the factors in the bone microenvironment that was responsible for the up-regulation of ezrin. The identification of PTHrP and ezrin as important regulators of lung cancer bone metastasis offers new mechanistic insights into the metastasis of lung cancer and provides potential targets for the prevention and treatment of lung cancer metastasis.

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Abbreviations

PTHrP:

Parathyroid hormone-related protein

HHM:

Humoral hypercalcemia of malignancy

ERM:

Ezrin-radixin-moesin

Luc:

Luciferase

YFP:

Yellow fluorescent protein

IC:

Intracardiac

SQ:

Subcutaneous

IVIS:

In vivo imaging system

RT-PCR:

Reverse transcription-polymerase chain reaction

B2M:

β2-microglobulin

siRNA:

Small interfering RNA

shRNA:

Short hairpin RNA

uPAR:

Plasminogen activator receptor, urokinase type

BMP-2:

Bone morphogenetic protein-2

FGF:

Fibroblast growth factor

TGF-β:

Transforming growth factor beta

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Acknowledgments

The authors would like to thank Drs. Wessel Dirksen and Ramiro Toribio, Dept. of Veterinary Biosciences, The Ohio State University, for critical reading of the manuscript and valuable comments on it. This work was supported by the National Institutes of Health, Nation Cancer Institute (RO1 CA 77911, PO1 CA100730) and the National Center for Research Resources (K26 RR00168).

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Authors and Affiliations

  1. Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH, 43210, USA

    Xiyun Deng, Sarah H. Tannehill-Gregg, Murali V. P. Nadella, Guangchun He, Andrea Levine & Thomas J. Rosol

  2. Bristol-Meyers Squibb, Pharmaceutical Research Institute, Evansville, IN, 47721, USA

    Sarah H. Tannehill-Gregg

  3. Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, Hunan, 410078, China

    Xiyun Deng, Guangchun He & Ya Cao

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  1. Xiyun Deng
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  2. Sarah H. Tannehill-Gregg
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Correspondence to Thomas J. Rosol.

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Conflict of Interest Statement: None declared.

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Deng, X., Tannehill-Gregg, S., Nadella, M.V.P. et al. Parathyroid hormone-related protein and ezrin are up-regulated in human lung cancer bone metastases. Clin Exp Metastasis 24, 107–119 (2007). https://doi.org/10.1007/s10585-007-9059-9

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  • DOI: https://doi.org/10.1007/s10585-007-9059-9

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