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SCUBE3 regulation of early lung cancer angiogenesis and metastatic progression

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Abstract

Signal peptide-CUB-EGF-like domain-containing protein 3 (SCUBE3) is strongly expressed in extremely invasive lung carcinoma. We showed in our previous study that SCUBE3 triggers the transforming growth factor-β pathway and subsequently promotes tumor angiogenesis and the epithelial–mesenchymal transition (EMT). However, the role of SCUBE3 in early tumor expansion hasn’t been fully demonstrated in vivo. The present study used dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to temporally assess tumor angiogenesis in SCUBE3-knockdown and control non-small-cell lung carcinoma (NSCLC) cancer cells in the early tumor stage (weeks 1–3). We further evaluated the metastatic potential of the SCUBE3-knockdown and control tumor cells using a circulating tumor cell (CTC) assay. The differences in gene expression profile between these cell lines were determined using microarray analysis. The results show that SCUBE3 knockdown was associated with lower vascular permeability in the tumor and effectively inhibited the metastatic potential of NSCLC, as evidenced by the decreased CTCs in the mice bearing SCUBE3-knockdown tumors. Microarray analysis revealed that several genes involved in angiogenesis and EMT were down-regulated in SCUBE3-knockdown tumors, including matrix metalloproteinases (MMPs) 2, 9, and 14, (MMP-2, MMP-9, and MMP-14, respectively), fibronectin (FN-1), lysyl oxidase (LOX), hairy/enhancer-of-split related with YRPW motif protein 1 (HEY1), early growth response protein 1 (EGR1), and interleukin 8 (IL-8). Together these data suggest that SCUBE3 is a potential target for pharmacological intervention. The findings of the present study also show that differences in vascular permeability precede the CTCs detection, indicating that DCE-MRI may be a sensitive biomarker for assessing tumor invasiveness.

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Abbreviations

AHNAK:

Neuroblast differentiation-associated protein AHNAK

ANGPTL4:

Angiopoietin-related protein 4

CTC:

Circulating tumor cell

CXCL10:

C-X-C motif ligand 10

DCE-MRI:

Dynamic contrast-enhanced magnetic resonance imaging

ECM:

Extracellular matrix

EGR1:

Early growth response protein 1

EMT:

Epithelial-mesenchymal transition

FN:

Fibronectin

FOV:

Field of view

GCOS:

GeneChip Operating System

Gd-DTPA:

Gadolinium-diethylenetriamine pentaacetic acid

H&E:

Hematoxylin and eosin

HEY1:

Hairy/enhancer-of-split related with YRPW motif protein 1

IL-8:

Interleukin 8

LOX:

Lysyl oxidase

MMP:

Matrix metalloproteinases

NEX:

Number of excitation

NOD-SCID:

Nonobese diabetic-severe combined immunodeficiency

NSCLC:

Non-small-cell lung carcinoma

PAI:

Plasminogen activator inhibitor

PBS:

Phosphate-buffered saline

SCUBE3:

Signal peptide-CUB-EGF-like domain-containing protein 3

Slth:

Slice thickness

SMAD:

Mothers against decapentaplegic homolog

T2WI:

T2-weighted images

TE:

Echo time

TGF-β:

Transforming growth factor-β

TR:

Repetition time

VEGF:

Vascular endothelial growth factor

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Acknowledgments

This paper was written in memory of late Dr. Konan Peck (1958–2011) and to honor his many valuable contributions to this research project. The authors acknowledge technical support from the Functional and Micro-Magnetic Resonance Imaging Center supported by the National Research Program for Genomic Medicine, National Science Council, Taiwan, Republic of China (NSC 99-3112-B-001-020). We thank Dr. Pan-Chyr Yang (Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan) for providing the CL1-5 cell lines with and without SCUBE3 knockdown. We also appreciate the help of Mr. Yi-Hua Hsu with discussions on histology.

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Authors and Affiliations

  1. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    Cheng-Hung Chou, Yi-Fang Cheng, Tiing Yee Siow, Amit Kumar, Konan Peck & Chen Chang

  2. Far Eastern Memorial Hospital, Pan-Chiao, Taipei, Taiwan

    Tiing Yee Siow

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  1. Cheng-Hung Chou
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  2. Yi-Fang Cheng
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Correspondence to Chen Chang.

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Cheng-Hung Chou and Yi-Fang Cheng contributed equally.

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Chou, CH., Cheng, YF., Siow, T.Y. et al. SCUBE3 regulation of early lung cancer angiogenesis and metastatic progression. Clin Exp Metastasis 30, 741–752 (2013). https://doi.org/10.1007/s10585-013-9575-8

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