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ORIGINAL ARTICLE
Year : 2018  |  Volume : 14  |  Issue : 12  |  Page : 1170-1172

Laminin is over expressed in breast cancer and facilitate cancer cell metastasis


Department of Thyroid and Breast Surgery, Clinical Medical College of Yangzhou University, Subei People's Hospital of Jiangsu Province, Yangzhou 225001, Jiangsu, P.R. China

Date of Web Publication11-Dec-2018

Correspondence Address:
Jiaxin Zhang
No. 98, Nantong West Road, Yangzhou 225001, Jiangsu
P.R. China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0973-1482.191035

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 > Abstract 


Aim of Study: Breast cancer invasion and metastasis is the main reason for the failure, and laminin is involved in it. This study intends to explore the expression of laminin in breast cancer and normal breast tissue and its clinical significance.
Materials and Methods: We use immunohistochemical assay for the detection of breast infiltrating ductal cancer tissues and normal breast tissues of laminin expression and discuss their role in breast cancer invasion and metastasis.
Results: Our results showed that laminin was positive expressed in normal breast tissue, and strongly positive expressed but lost its' continuity in the breast cancer tissue.
Conclusion: This results revealed laminin is involved in breast cancer invasion and metastasis, and we can use this to determine whether the integrity of a basement membrane for differential diagnosis of benign and malignant breast tumors.

Keywords: Basement membrane, breast neoplasms, immunohistochemistry, laminin


How to cite this article:
Qiu X, Tan H, Fu D, Zhu Y, Zhang J. Laminin is over expressed in breast cancer and facilitate cancer cell metastasis. J Can Res Ther 2018;14, Suppl S5:1170-2

How to cite this URL:
Qiu X, Tan H, Fu D, Zhu Y, Zhang J. Laminin is over expressed in breast cancer and facilitate cancer cell metastasis. J Can Res Ther [serial online] 2018 [cited 2019 Sep 21];14:1170-2. Available from: http://www.cancerjournal.net/text.asp?2018/14/12/1170/191035




 > Introduction Top


Tumor cell invasion and metastasis is a complex process steps, and a variety of cytokines and molecules involved. The interaction between cancer cells and basement membrane and break through the basement membrane is the key step, which involves many kinds of cell factors, intracellular protein expression, and cell surface receptor expression. Our study detected laminin expression level in breast cancer and normal breast tissues by immunohistochemical method in order to discuss the role of laminin in breast cancer invasion and metastasis.


 > Materials and Methods Top


Case data

We collected the patients' breast tissues from November 2011 to 2016 April in our hospital who underwent surgical treatment, including 63 cases of breast infiltration ductal carcinoma and 70 cases of normal breast tissues. All patients did not accept radiotherapy or chemotherapy treatment before operation. All cases were preceded with modified radical mastectomy. The tumor tissue was taken from the center of the nonnecrotic tumor. The normal breast tissue was taken from breast with the distance from the tumor to the minimum distance larger than 3 cm. The diagnosis was confirmed by two senior pathologists with hematoxylin and eosin staining. The expression of laminin in the two groups was detected by immunohistochemistry method.

Main reagent

Mouse antihuman laminin monoclonal antibody (ready to use type), MaxVision enzyme labeled goat-antimouse IgG antibody (horseradish peroxidase-polymer goat-antimouse IgG, ready to use type), and 3,3' diaminobenzidine (DAB) chromogenic reagents were purchased from Fuzhou Maixin Biotech Co., Ltd.

Methods

Breast tissue samples were selected by an experienced pathologist based on diagnosis and microscopic morphology; tissue sections were deparaffinized with xylene, then rehydrated in ethanol washes (100%, 90%, 80%, and 70%) and then washed with phosphate-buffered saline (PBS) and antigen retrieval through microwave oven for 20 min. Next, samples were washed with PBS and treated with 3% H2O2 for 15 min to block endogenous peroxidase activity and then washing with PBS, incubated with 10% goat serum for 1 h to block nonspecific bindings. Samples were then incubated with a 1:400 dilution of mouse antihuman laminin monoclonal antibody overnight at 4°C in a humidified chamber. Then rinsed with PBS and incubated with MaxVision enzyme labeled goat-anti-mouse IgG antibody (1:100) for 1 h at room temperature. Washings with PBS, and then incubated for 3–5 min with DAB substrate and 3-amino-9-ethylcarbazole substrate separately, followed by counterstaining with Meyer's hematoxylin for 30 s. Coverslips, examined and captured the slides under a microscope with a digital camera.

Result judgment

Criterion of laminin staining: Colorless was negative, pale brown was positive, brownish black was strong positive.

Statistical methods

SPSS (Statistical Package for the Social Science; SPSS Inc., Chicago, IL, USA) version 19 for Microsoft Windows was used for statistical analysis. The expression of laminin in breast cancer and normal breast tissue was tested by Fisher's exact probability method, and the test level was α = 0.01.


 > Results Top


In normal breast tissue, the positive and strong positive expression rate of laminin was 70/70 (100%), 0/70 (0%), respectively. In breast invasive ductal carcinoma, the positive and strong positive expression rate of laminin was 11/63 (17.5%), 52/63 (82.5%), respectively [Figure 1] and [Table 1]. Comparison between the two groups, the difference was statistically significant (P < 0.01).
Figure 1: The expression of laminin in breast invasive ductal carcinoma and normal tissues. (a and b) laminin was positive expressed in normal breast tissue, (×200); (c) laminin was positive expressed in invasive ductal carcinoma, (×100); (d) laminin was strongly positive expressed in invasive ductal carcinoma, (×200)

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Table 1: Expression of laminin in normal breast tissue and invasive ductal carcinoma

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 > Discussion Top


Basement membrane is recognized as a cancer cell defense barrier, which inhibits the local infiltration of cancer cells by inhibiting the adhesion of cancer cells to it. The cancer cell breaks through the basement membrane which is the key step in the process of tumor development, and it is the important factor to judge the malignant degree of tumor in clinical and scientific research. Laminin is a kind of glycoprotein[1],[2],[3] extracted from the basement membrane by Timpl et al., which was first extracted from mouse Engelbreth-Holm-Swarm sarcoma in 1979. It is a structural component of basement membrane, which is expressed in normal basement membrane by immunohistochemistry. Laminin is a key extracellular matrix regulation factor involved in cell adhesion, migration, differentiation, proliferation, and therefore in the maintenance of tissue morphology and internal environment stable, matrix-mediated signal transduction, tumor local invasion, and metastasis.[4],[5],[6],[7],[8]

Many epithelial cancers, including lung cancer, colorectal cancer, squamous cell carcinoma, and so on, are highly expressed laminin, especially its gamma 2 chain, and it is closely related to tumor invasion and clinical prognosis.[9],[10],[11] The matrix metalloproteinases family (MMPs) can expose the hidden splice sites of the breast cancer cells through the degradation of laminin, thereby stimulating the breast cancer cells, so as to enhance the migration ability of cancer cells.[12],[13],[14]

Immunohistochemical method was used for the detection of laminin protein expression in normal mammary gland and breast cancer, we found that in normal breast tissue, positive expression of laminin in the basement membrane around the mammary gland, and as a continuous linear expression. And in most breast cancer tissues, laminin was strongly positive, and break through the basement membrane, loss of continuity; in the positive expression of tissue, the basement membrane is still a part of the integrity [Figure 1]c. This showed that the expression of laminin was enhanced and the integrity of basement membrane was damaged in the breast malignant tumor. We therefore speculate: In breast cancer tissues, cancer cells through enhanced expression of laminin, make it easier to migrate and adhere to the basement membrane, and secretion of MMP-2 enzyme substrate degradation of the basement membrane and extracellular matrix, which is conducive to the invasion and metastasis of the cancer cells of local. This based on three points: (1) Breast cancer tissues of laminin enhanced expression was significantly higher than that of normal breast tissue; (2) laminin is interact with MMP-2 and other important matrix enzyme, and MMP-2 is an important enzyme for the degradation of the extracellular matrix and basement membrane; (3) with the enhanced expression of laminin, the integrity of basement membrane was destroyed in breast cancer tissues.

Based on the above experimental results and inferences, it is concluded that the expression of laminin can be used to judge the integrity of basement membrane and the auxiliary diagnosis of breast cancer. In clinical work, it is sometimes difficult to judge whether the invasion of the basement membrane. According to laminin in the basement membrane of normal breast tissue continuous expression and in cancer tissue which destroy the continuity and strongly positive in expression can be of pathologic diagnosis of difficult cases by laminin immunohistochemical detection, to identify breast cancer and benign diseases. For example, in the fast frozen section of the breast, when it is difficult to diagnose, it can be used to detect the expression of laminin in order to judge whether it is malignant or not.


 > Conclusion Top


Laminin is over expressed in breast cancer, and it is involved in breast cancer invasion and metastasis.

Acknowledgment

We want to take this chance to thanks to Doctor Changren Zhu, a kindly pathologist of Subei People's Hospital of Jiangsu Province. In the process of composing this paper, he gives us many academic and constructive advices, and helps us to correct our paper.

Financial support and sponsorship

Our study was supported by the Fund of Subei People's Hospital of Jiangsu Province (No. yzucms201202) and the National Natural Science Foundation of China (No. 81172508).

Conflicts of interest

There are no conflicts of interest.



 
 > References Top

1.
Ning L, Kurihara H, de Vega S, Ichikawa-Tomikawa N, Xu Z, Nonaka R, et al. Laminin a1 regulates age-related mesangial cell proliferation and mesangial matrix accumulation through the TGF-ß pathway. Am J Pathol 2014;184:1683-94.  Back to cited text no. 1
    
2.
Xu J, Li D, Li X, Liu Z, Li T, Jiang P, et al.67 laminin receptor promotes the malignant potential of tumour cells up-regulating lysyl oxidase-like 2 expression in cholangiocarcinoma. Dig Liver Dis 2014;46:750-7.  Back to cited text no. 2
    
3.
Timpl R, Rohde H, Robey PG, Rennard SI, Foidart JM, Martin GR. Laminin – A glycoprotein from basement membranes. J Biol Chem 1979;254:9933-7.  Back to cited text no. 3
    
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Tzu J, Marinkovich MP. Bridging structure with function: Structural, regulatory, and developmental role of laminins. Int J Biochem Cell Biol 2008;40:199-214.  Back to cited text no. 4
    
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Pal S, Moulik S, Dutta A, Chatterjee A. Extracellular matrix protein laminin induces matrix metalloproteinase-9 in human breast cancer cell line mcf-7. Cancer Microenviron 2014;7:71-8.  Back to cited text no. 5
    
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Denoyer D, Kusuma N, Burrows A, Ling X, Jupp L, Anderson RL, et al. Bone-derived soluble factors and laminin-511 cooperate to promote migration, invasion and survival of bone-metastatic breast tumor cells. Growth Factors 2014;32:63-73.  Back to cited text no. 6
    
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Giannelli G, Pozzi A, Stetler-Stevenson WG, Gardner HA, Quaranta V. Expression of matrix metalloprotease-2-cleaved laminin-5 in breast remodeling stimulated by sex steroids. Am J Pathol 1999;154:1193-201.  Back to cited text no. 7
    
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Kusuma N, Denoyer D, Eble JA, Redvers RP, Parker BS, Pelzer R, et al. Integrin-dependent response to laminin-511 regulates breast tumor cell invasion and metastasis. Int J Cancer 2012;130:555-66.  Back to cited text no. 8
    
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Yamamoto H, Itoh F, Iku S, Hosokawa M, Imai K. Expression of the gamma(2) chain of laminin-5 at the invasive front is associated with recurrence and poor prognosis in human esophageal squamous cell carcinoma. Clin Cancer Res 2001;7:896-900.  Back to cited text no. 9
    
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Sun L, Liu L, Liu X, Wang Y, Li M, Yao L, et al. Gastric cancer cell adhesion to laminin enhances acquired chemotherapeutic drug resistance mediated by MGr1-Ag/37LRP. Oncol Rep 2014;32:105-14.  Back to cited text no. 10
    
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Yang ZY, Zhou XS, Wei X, Liu Y, Gong C, Chen G, et al. MYC suppresses ovarian cancer cell adhesion and invasion ability through downregulation of human integrin β1. J Pract Med 2014;7:1048-52.  Back to cited text no. 11
    
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Giannelli G, Falk-Marzillier J, Schiraldi O, Stetler-Stevenson WG, Quaranta V. Induction of cell migration by matrix metalloprotease-2 cleavage of laminin-5. Science 1997;277:225-8.  Back to cited text no. 12
    
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Koshikawa N, Minegishi T, Sharabi A, Quaranta V, Seiki M. Membrane-type matrix metalloproteinase-1 (MT1-MMP) is a processing enzyme for human laminin gamma 2 chain. J Biol Chem 2005;280:88-93.  Back to cited text no. 13
    
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Wang HY, Zhang XB, Wang M. Expression of matrix metalloproteinase 9 (MMP-9) and laminin-receptor in breast carcinoma and their correlation with tumor metastasis and prognosis. Ai Zheng 2003;22:529-32.  Back to cited text no. 14
    


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