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ORIGINAL ARTICLE  
Year : 2011  |  Volume : 54  |  Issue : 3  |  Page : 509-513
Hepatocyte growth factor profile with breast cancer


Department of Chemical Pathology and Pathology, Medical Research Institute, Alexandria University, Egypt

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Date of Web Publication20-Sep-2011
 

   Abstract 

Background: The multifunctional hepatocyte growth factor (HGF) is the ligand of c-Met receptor; it plays important role in mammary differentiation. HGF-Met signaling is a critical downstream function of c-Src-Stat3 pathway in mammalian tumorigenesis. Aim: Evaluation of tissue c-Met receptor hepatocyte growth factor receptor (HGFR) and serum level of HGF in female breast ductal carcinoma. Materials and Methods: Sixty-eight premenopausal females were divided as 30 control females subdivided into: [Group 1] 15 healthy volunteer females and [Group 2] five with fibrocystic disease and 10 having fibroadenoma of the breast and patients group [Group 3] consisted of 38 female patients with breast ductal carcinoma. Thorough clinical examination, preoperative fine needle aspiration cytology, estimation of fasting serum glucose, urea, creatinine, and uric acid levels, alanine aminotransferase activities, C-reactive protein, HGF level, before surgery and histopathological examination of the breast masses, and immunohistochemical detection of HGFR were done. Results and Conclusions: Significant increase in serum HGF levels were found in patients with breast cancer as compared with controls. Significant increase was also seen in patients with breast cancer with and without lymph node metastasis when each subgroup was compared with controls. Serum level of HGF is an independent prognostic indicator of breast cancer. Fibrocystic disease of the breast showed weak HGFR expression, while in normal tissue, HGFR was scanty; meanwhile, breast invasive ductal carcinoma showed homogenous strong reaction to HGFR. HGF is only one of a number of key factors involved in breast cancer and preoperative high serum HGF levels and malignancy occur usually together.

Keywords: Breast lumps, hepatocyte growth factor, immunohistochemistry, prognostic parameters

How to cite this article:
EL-Attar HA, Sheta MI. Hepatocyte growth factor profile with breast cancer. Indian J Pathol Microbiol 2011;54:509-13

How to cite this URL:
EL-Attar HA, Sheta MI. Hepatocyte growth factor profile with breast cancer. Indian J Pathol Microbiol [serial online] 2011 [cited 2018 Aug 22];54:509-13. Available from: http://www.ijpmonline.org/text.asp?2011/54/3/509/85083



   Introduction Top


Acquisition of an autocrine growth-stimulating circuit is a hallmark of cancer cells. [1] Growth factors (GFs) are polypeptide molecules that bind to receptors on cell surface. [2] Hepatocyte growth factor (HGF) is an important paracrine mediator of mesenchymal-epithelial interaction, instructing epithelial cells to undergo processes such as cell proliferation, survival, migration, and morphogenesis. [1]

HGF, also known as Scatter factor, Hepatopoietin A, lung fibroblast derived mitogen, and Met-ligand, was originally discovered with liver regeneration. [3]

HGF is synthesized as biologically inactive chain precursor (728 amino acids called pro-HGF) [4] which requires proteolytic processing, by various serine proteases to generate active two-chain form (HGF) in vitro. [5]

HGF is primarily expressed in stromal cells and weakly in epithelial and endothelial cell. [6]

Activator inhibitor type-1 and type-2 are two serine protease inhibitors with the ability to bind HGF activator and thus block its HGF-activating properties. [1]

HGF promotes mammary ductal morphogenesis and in maintaining ductal spacing. [7] Also, c-Met and Erb-B 2 are associated with morphogenic and functional differentiation of normal mammary gland epithelium. [8]

HGF is expressed in "benign breast disease" and detected as incidental microscopic findings. [9]

Fibrocystic changes are the most frequent benign disorder of the breast. [8] Cysts contain a variety of biologically active substances including steroids, cytokines, GFs, and proteins. [10]

Hepatocyte GH-Met signaling is a critical downstream function of c-Src-Stat3 pathway in mammalian tumorigenesis. [8] Through Met receptor binding, HGF aberrantly activates the motility/chemoinvasion, proliferation/survival, and apoptosis that characterize the epithelial-mesenchymal transition of aggressive carcinomas. [11]

There is still limited knowledge about the role of HGF in breast cancer, [12] and it might reflect the severity of cancer invasion which deserve further evaluation.

Aim

Evaluation of tissue c-Met receptor (HGFR) and serum level of HGF in females with breast ductal carcinoma.


   Materials and Methods Top


The study was done in accordance to ethical standards of Medical Research Institute based on Helsinki Declaration revised in 2000. Sixty-eight adult premenopausal females, recruited consecutively, after takingtheir consents, were divided as: Group 1 comprised of 30 control females; 15 normal volunteer females, five with fibrocystic disease of the breast and 10 having breast fibroadenoma and Group 2 comprised of 38 females with breast ductal carcinoma. All the patients had undergone a thorough clinical examination and fine needle aspiration cytology for breast lump, if any. Data on lymph node status, primary tumor staging according to TNM classification, estrogen and progesterone receptors status, and histological grading of carcinoma were recorded. Before surgery, laboratory investigations in serum included fasting glucose (FSG), urea (Ur), creatinine (Cr), and uric acid (UA) levels, alanine aminotransferase (ALT) activities, and HGF. [13] Histopathological examination of the breast mass after surgical removal and immunohistochemistry for HGFR by avidin-biotin staining technique using R and D system, according to manufacturer instructions (catalog number AF276), were done.

Immunohistochemistry


The formalin-fixed, paraffin-embedded blocks were cut in 3 to 5 mirometers thin sections, brought onto charged slides which were dehydrated, stained immunohistochemically by avidin-biotin staining technique using R and D system, according to manufacturer instructions, the slides were incubated with HGF receptor (catalog number AF276) 10 microg/ml overnight at 4°C, preceded by heat-induced epitope retrieval. DAB chromogen was used and counterstained with hematoxylin. The reaction product appeared as cytoplasmic and cell membranous brown stain. The reaction is semiquantitatively scored in 4 scale system (0 = negative, + = mild, ++ = moderate, and +++ = marked reaction). Statistical analyses were done using SPSS software package version 17 (SPSS, Chicago, IL). Presentation was in the form of median and range and Spearman's rank correlation coefficient. A "P" value <0.05 was considered statistically significant.


   Results Top


The median value of age in the studied groups is shown in [Table 1]. A significant decrease in age was noticed in patients with benign breast lumps group when compared with control group. A significant increase in age was found in the entire breast cancer patients' group and in patients with and without lymph node (LN) metastasis group when each was compared with its value in patients with benign breast lumps group.
Table 1: Age distribution (median and range) of the studied groups

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Serum level of FSG, Ur, Cr, UA, and serum ALT and serum alkaline phosphatase activities were within normal limits in the studied groups.

HGF was 554 pg/ml (279-1 220 pg/ml) in patients with benign breast lumps [Group 2], 447.5 pg/ml (279-770 pg/ml) in patients with cystic disease [Group 2a] and was 580 pg/ml (353-1 220 pg/ ml) in patients with fibroadenoma [Group 2b]. However, its level was 1 073.5 pg/ml(356-2 352 pg/ml) in the breast cancer patients' group [Group 3] showing significant increase in patients with benign breast lumps group and in patients with fibroadenoma and breast cancer patients group when each was compared with control group [Table 2]. A significant increase in HGF level was also found in breast cancer patients group when compared with patients having benign breast lumps and subjects with cystic disease of the breast and subjects with fibroadenoma of the breast when each was compared with patients with benign breast lumps group [Table 2].
Table 2: Serum values of hepatocyte growth factor (Pg/ml) in the studied groups

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No significant difference in serum level of HGF and the prognostic parameters in breast cancer patients was observed. This is shown in [Table 3].
Table 3: Statistical difference in HGF in relation to age, tumor size, grade, lymph nodes involvement, stage, ER, PR, and angiogenesis in whole breast cancer patients group

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Tissue immunohistochemistry reveals that fibrocystic disease of the breast showed weak HGFR expression, while in normal tissue in the surroundings, HGF expression was scanty; meanwhile, invasive ductal carcinoma of the breast showed marked reaction to HGF [Figure 1] and [Figure 2].
Figure 1: Fibrocystic disease of the breast showing weak HGFR expression (IHC-HGF x100), while in normal tissue in the surroundings, HGF expression is scanty

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Figure 2: Invasive ductal carcinoma of the breast showing homogenous strong reaction to HGFR (IHC-HGF x40)

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   Discussion Top


HGF which belongs to the plasminogen prothrombin gene superfamily is one of the several GFs that have been shown to play an active role in mammary gland development. [1],[7],[14] Genetic aberration in GF signaling pathways are strongly connected with lumps. [1]

Sattler and Salgia [15] found that chronic or dysregulated activation of Met/HGF pathway may lead to increased cell growth, invasion, angiogenesis, metastasis, reduced apoptosis, altered cytoskeletal functions, and other biological changes and HGF and its receptor is one of a number of key factors involved in cancer.

In the present work, the median value of HGF was 311 pg/ml (177-814 pg/ml in control group [Table 2], which was comparable with the level found by previous workers.[12]

In the present study, HGF level in breast cancer patients' group was 1 073.5 pg/ml (356-2 352 pg/ml) [Table 2]. This was not in agreement with results of previous workers [12] but comparable with the work done by Taniguchi et al. [16] There was a significant increase in HGF value in breast cancer patients group when compared with control group P = 0.000) [Table 2]. This finding is in accordance with the results of other workers. [4],[12],[15],[17],[18],[19]

Previous workers found that HGF is higher in tumor tissues than in adjacent normal tissue. Deregulated HGF/Met system triggers a program for epithelial-mesenchymal transition conferring growth of carcinoma. [2],[4],[20],[21],[22]

Immunohistochemical analysis of biopsy samples of human breast carcinoma indicated that in normal mammary gland, c-Met is localized in the ductal epithelium. The level of expression of c-Met in primary carcinomas was maintained in some cases and in other cases was elevated. Frequently, there was evidence of heterogeneity in cellular expression of c-Met within individual tumors suggesting that microenvironmental factors may regulate receptors expression. [22] Invasion occurs within a tumor-host microecology, when stroma and tumor cells exchange enzymes and cytokines that modify the extracellular matrix, stimulates migration, and promotes proliferation and survival. [23]

In the present work, immunohistochemical analysis showed that HGFR concentration was significantly highly expressed in invasive ductal carcinoma of the breast tumor tissues than benign lumps and normal tissues [Figure 1] and [Figure 2].

The increase in HGF could be explained by overstimulation of the cells that secrete HGF through autosecretion or mutation and aberrant regulation of the HGF/Met signaling pathway or an imbalance between HGF activators and inhibitors. [20]

It has been found that elevated HGF content in tumor tissue may predict more aggressive biology in non-small-cell lung cancer patients but still limited knowledge about the role of HGF in breast cancer. [12],[24]

In the present work, no significant difference was found in serum HGF level between patients having breast cancer with and without lymph nodes (LN) metastasis [Figure 1] as HGF median value was 1 069 pg/ml (356-2 244 pg/ml) in patients having breast cancer with LN metastasis group and was 1 078 pg/ml (491-2 352 pg/ml) in patients without LN metastasis group. No significant difference in HGF level and LN metastasis was also found by Sung et al. [25] However, in a previous work, patients in more advanced TNM staging were shown to have higher serum HGF. [12]

In the current study, in breast cancer patients, no significant difference was found in HGF and age, tumor size, and tumor grade [Table 3] like others.[12],[15],[26],[27],[28] However, contrary to our findings, higher serum levels of HGF were found in patients with high-grade tumors. [12],[15] The correlation of HGF level with staging, in our patients, cannot be found as they were seeking medical advice before reaching this late stage [Table 3].

Sheen-Chen et al.[12] found that there were significantly higher serum level of HGF in patients with negative estrogen receptors, in patients with poorly differentiated tumors, and in patients with more advanced LN status, in patients with distant metastasis and with more advanced TNM staging.

In the present study, no significant associations in HGF level and hormone receptors (ER and PR) were found [Table 3] which goes in agreement with other work[24] also between tissue level of HGF in breast cancer tissue and hormone receptors. [26],[27]

In the present study, serum level of HGF did not show significant difference with angiogenesis [Table 3]. Previous researchers[24] found no significant associations. However, other workers found that HGF stimulates tumor invasion and revascularization. [12],[16] So, it may play important roles in tumor growth and metastasis.

The pivotal role of HGF in cancer has indicated it as being a potential target to cancer therapy. [29]


   Conclusions Top


Serum level of HGF in breast cancer patients showed significant increase when compared with controls. HGF is an independent prognostic marker in breast cancer. HGFR expression is weak in fibrocystic disease of the breast, while in invasive ductal carcinoma of the breast, HGFR expression is marked.


   Recommendations Top


There is still limited knowledge about the role of HGF in breast cancer, and more work is needed to elucidate the possible relationship between inhibition of HGF/MET pathway as a therapeutic strategy in breast malignancies.


   Acknowledgment Top


This research did not receive any specific grant from any funding agency in the public, commercial, or not-for-profit sector.

 
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Correspondence Address:
Hoda A EL-Attar
Department of Chemical Pathology, Medical Research Institute, Alexandria University
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0377-4929.85083

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