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SMAD4 is a potential prognostic marker in human breast carcinomas

  • Research Article
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Tumor Biology

Abstract

SMAD4 is a downstream mediator of transforming growth factor beta. While its tumor suppressor function has been investigated as a prognostic biomarker in several human malignancies, its role as a prognostic marker in breast carcinoma is still undefined. We investigated SMAD4 expression in breast carcinoma samples of different histologic grades to evaluate the association between SMAD4 and outcome in breast cancer. We also investigated the role of SMAD4 expression status in MDA-MB-468 breast cancer cells in responding to TGF-β stimulation. SMAD4 expression was assessed in 53 breast ductal carcinoma samples and in the surrounding normal tissue from 50 of the samples using immunohistochemistry, Western blot, and real-time PCR. TGF-β-SMAD and non-SMAD signaling was assessed by Western blot in MDA-MB-468 cells with and without SMAD4 restoration. SMAD4 expression was reduced in ductal breast carcinoma as compared to surrounding uninvolved ductal breast epithelia (p < 0.05). SMAD4 expression levels decreased from Grade 1 to Grade 3 ductal breast carcinoma as assessed by immunohistochemistry (p < 0.05). Results were recapitulated by tissue array. In addition, immunohistochemistry results were further confirmed at the protein and mRNA level. We then found that non-SMAD MEK/MAPK signaling was significantly different between SMAD4 expressing MDA-MB-468 cells and SMAD4-null MDA-MB-468 cells. This is the first study indicating that SMAD4 plays a key role in shifting MAPK signaling. Further, we have demonstrated that SMAD4 has a potential role in the development of breast carcinoma and SMAD4 was a potential prognostic marker of breast carcinoma. Our findings further support the role of SMAD4 in breast carcinoma development. In addition, we observed an inverse relationship between SMAD4 levels and breast carcinoma histological grade. Our finding indicated that SMAD4 expression level in breast cancer cells played a role in responding non-SMAD signaling but not the canonic SMAD signaling. Further mechanistic studies are necessary to establish the role of SMAD4 in breast carcinoma prognosis and potential specific targeting.

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Abbreviations

TGF:

β: Transforming growth factor beta

MAPK:

Mitogen-activated protein kinase

PI3K:

Phosphatidylinositol 3-kinase

ERK:

Extracellular signal-regulated kinase

MMP:

Matrix metalloproteinase

EMT:

Epithelial–mesenchymal transformation

IHC:

Immunohistochemistry

ERα:

Estrogen receptor α

AKT:

Protein kinase B

JNKs:

c-Jun amino-terminal kinases

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Acknowledgments

Portions of this work were supported by the National 863 Program (no. 2004AA205020) and the National Natural Science Foundation of China (no. 81272243) grants, as well as pilot funds from the UAB Breast Cancer NCI SPORE project and NIH CA 108741 grant.

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Authors and Affiliations

  1. The Key Laboratory of Pathobiology, Ministry of Education, Jilin University, Changchun, 130021, China

    Nan-nan Liu, Yue Xi, Qinghua Zeng & Yu-lin Li

  2. Department of Pediatrics, Wayne State University School of Medicine, Detroit, MI, 48201, USA

    Yue Xi, Michael U. Callaghan, Andrew Fribley & Qinghua Zeng

  3. Division of Hematology/Oncology, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Lakisha Moore-Smith, Jacquelyn W. Zimmerman, Boris Pasche & Qinghua Zeng

  4. Department of Pathology, College of Basic Medicine , Beihua University, Jilin, 132013, China

    Nan-nan Liu

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  1. Nan-nan Liu
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Correspondence to Qinghua Zeng or Yu-lin Li.

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Nan-nan Liu and Yue Xi These authors contributed equally to this work.

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Figure 1

Microarray SMAD4 immunohistochemistry staining in breast carcinoma and surrounding normal breast epithelia. a Slide IMH-371 (Human breast: cancer-normal adjacent) and IMH-364 (Human breast: cancer-metastasis-normal) from IMGENEX were stained with anti-SMAD4 antibody (1:200). b There was a total of 70 cases of breast carcinoma on the two microarray slides and 39 cases with matched normal breast epithelia. Overall, SMAD4 expression was significantly lower in breast carcinoma (IRS = 4.35 ± 4.11) than that in the normal breast epithelia (IRS = 10.92 ± 1.96; P < 0.05) (PPTX 2325 kb)

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Liu, Nn., Xi, Y., Callaghan, M.U. et al. SMAD4 is a potential prognostic marker in human breast carcinomas. Tumor Biol. 35, 641–650 (2014). https://doi.org/10.1007/s13277-013-1088-1

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  • DOI: https://doi.org/10.1007/s13277-013-1088-1

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