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ORIGINAL ARTICLE
Year : 2017  |  Volume : 13  |  Issue : 5  |  Page : 833-836

The clinical significance of secreted protein acidic and rich in cysteine expression in breast cancer tissue and its association with prognosis


1 Department of Oncology Surgery, First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, China
2 Department of Oncology Gynecology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, China

Date of Web Publication13-Dec-2017

Correspondence Address:
Suyang Guo
Department of Oncology Gynecology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233000
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jcrt.JCRT_424_17

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 > Abstract 


Objective: The aim of this study was to evaluate the secreted protein acidic and rich in cysteine (SPARC) expression in breast cancer tissue and its association with patients' clinical pathology characteristics and prognosis.
Materials and Methods: Eight-eight cases with confirmed diagnosis of breast cancer who received operation from January 2010 to February 2016 were included in this study. The SPARC expression in cancer tissue was examined by immunohistochemical method. The SPARC expression status, clinical pathology characteristics, and prognosis of included patients were recorded and evaluated.
Results: SPARC protein was mainly expressed in cytoplasm and stroma of tumor tissue with dark brown and purple stain. The SPARC protein-positive expression rate was 69.3% (61/88) in cancer tissue. The positive expression of SPARC in breast tissues was not significantly correlated with age, menstruation status, tumor, node, and metastasis stage, tumor size, progesterone level, and HER-2 status (P > 0.05). However, SPARC protein-positive expression was correlated with tumor differentiation (P < 0.05), estrogen receptor expression (P < 0.05), and lymph node metastasis (P < 0.05). The 3-year disease-free survival (DFS) was 60.8% and 71.2% for SPARC-positive and -negative groups with no statistical difference (P > 0.05); there was no statistical difference of disease progression risk between the SPARC-positive and -negative groups (hazard ratio = 1.78, 95% confidence interval: 0.80–3.57, P > 0.05). However, SPARC-positive and -negative patients have shown a trend of DFS difference.
Conclusion: SPARC is closely related to the development of breast cancer and can be used as a tumor marker for breast cancer recurrence.

Keywords: Breast neoplasms, clinical and pathological features, immunohistochemistry, prognosis, secreted protein acidic and rich in cysteine


How to cite this article:
Guo W, Zhang M, Chen Y, Guo S. The clinical significance of secreted protein acidic and rich in cysteine expression in breast cancer tissue and its association with prognosis. J Can Res Ther 2017;13:833-6

How to cite this URL:
Guo W, Zhang M, Chen Y, Guo S. The clinical significance of secreted protein acidic and rich in cysteine expression in breast cancer tissue and its association with prognosis. J Can Res Ther [serial online] 2017 [cited 2019 Nov 22];13:833-6. Available from: http://www.cancerjournal.net/text.asp?2017/13/5/833/220477




 > Introduction Top


Breast cancer is the most common type of solid malignant tumor and is the cancer with the highest incidence rate among women.[1] Epidemiological data from the North American authorities show that in 2013, 23,458 new breast cancer cases and 40,030 deaths from breast cancer were reported in the United States.[2],[3] Breast cancer is the malignancy with the highest incidence and mortality rates next to lung cancer. Although the incidence rate of breast cancer remains high, breast cancer patients exhibit improved prognosis and high 5-year survival rates because of the development of targeting drugs over the past few years. Breast cancer, however, still lacks effective prognostic biomarkers.

The secreted protein acidic and rich in cysteine (SPARC), also known as osteonectin and basal membrane-40 protein, is a 32-kDa secreted glycoprotein.[4] SPARC interacts with the extracellular matrix (ECM) protein, promotes cell adhesion, and induces cell migration.[5],[6] SPARC is also implicated in tissue remodeling, angiogenesis,[7],[8] embryogenesis, and tumorigenesis.[9],[10] However, the current mechanism of SPARC protein expression in tumor cells, the specific mechanism of interaction between SPARC and ECM, and the impact of SPARC on breast cancer prognosis remain unclear. The studies have recently shown that SPARC is associated with the development and prognosis of various tumors, such as gastric and colorectal cancers. However, few studies have reported on the relationship between the expression levels of SPARC with the clinical features and prognosis of breast cancer. In this study, we utilized the immunohistochemical SP method to detect SPARC expression in breast cancer tissues and to identify its relationship with the clinical features and prognosis of breast cancer. Moreover, we explored the role of SPARC in the development and progression of colorectal cancer and its feasibility as a prognostic marker.


 > Materials and Methods Top


Breast cancer patients

We recruited 87 breast cancer patients who underwent surgical treatment and who had preserved complete tumor tissue blocks at the Surgical Oncology of the First Affiliated Hospital of Bengbu Medical College from January 2010 to March 2016. All of the 87 patients were female whose ages ranged between 25 and 78 with an average of 50.6 ± 12.8. The inclusion criteria were as follows: Diagnosed with breast cancer based on the postoperative pathological section, complete clinical follow-up survey, older than 18 years old, at a clinical stage between I and II, and never received preoperative chemoradiotherapy or biological immunotherapy. The exclusion criteria were as follows: Younger than 18 years old, incomplete clinical follow-up data, had other malignant tumors at the same time, and received preoperative chemoradiotherapy.

Equipment and reagents

Paraffin slicing machine purchased from Leica, Germany (RM2235); microscope purchased from Olympus, Japan (OLYMPUSBX-40); electric thermostat manufactured by Shanghai Rongke Detection Instrument Co., Ltd., (DHG-9035A); low-speed centrifuge purchased from Beckman Coulter, Germany (Allegra X-12). Mouse antihuman SPARC monoclonal antibody (working concentration 1:500) purchased from the British Abcam; immunohistochemical SP kit; biotin-labeled goat anti-mouse/rabbit IgG (secondary antibody); DAB kit and citric acid repair liquid (100 times) purchased from Fuzhou Maixin New Biotechnology Development Co., Ltd.

Secreted protein acidic and rich in cysteine immunohistochemical detection

The immunohistochemical SP method was used to test the expression levels of SPARC in breast cancer tissue and was performed in strict accordance with the provided instructions. PBS replaced primary antibodies in the negative control. A SPARC-positive section of known breast cancer tissue was used as the positive control. Specific steps are as follow: (1) dewaxing and hydration, (2–3) immunohistochemical staining, and (4) sealing for microscopy examination.

Determination of positive results

The double-blind method was used to determine the staining results. The total numbers of cells and SPARC-positive cells from 5 fields of view from each slide were counted under the microscope at ×40 magnification. A total of 100 cells from each field of view were scored based on the percentage of positive cells: 0 point, negative; 1 point, positive cells no >10%; 2 points, 11%–50%; 3 points, 51%–75%; and 4 points, >75%. If the product of staining intensity and the percentages of positive cells were not <3, the immune reaction was positive; otherwise it was negative.

Statistical analysis

All statistical analyses were done by STATA 12.0 software (https://www.stata.com/), the measurement data were expressed by ̄ ± s and compared by t-test. Count data were expressed with n (%) and analyzed Chi-square test or Fisher's exact test. Log-rank test was used to compare the survival between SPARC-positive and -negative groups. P < 0.05 was considered as statistically significant.


 > Results Top


Secreted protein acidic and rich in cysteine protein expression in tumor tissue

SPARC protein was mainly expressed in cytoplasm and stroma of tumor tissue with dark brown and purple stain [Figure 1]. The SPARC protein-positive expression rate was 69.3% (61/88) in cancer tissue.
Figure 1: Secreted protein acidic and rich in cysteine expression in breast cancer tissue (a) secreted protein acidic and rich in cysteine-positive expression; (b) secreted protein acidic and rich in cysteine-negative expression)

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Secreted protein acidic and rich in cysteine protein expression and patients clinical characteristics

The positive expression of SPARC in breast tissues was not significantly correlated with age, menstruation status, tumor, node and metastasis stage, tumor size, progesterone level, and HER-2 status (P > 0.05). However, SPARC protein-positive expression was correlated with tumor differentiation (P < 0.05), estrogen receptor expression (P < 0.05), and lymph node metastasis (P < 0.05) [Table 1].
Table 1: The secreted protein acidic and rich in cysteine protein expression and patients clinical characteristics, n (%)

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Secreted protein acidic and rich in cysteine protein expression and prognosis

The 3-year disease-free survival (DFS) was 60.8% and 71.2% for SPARC-positive and -negative groups with no statistical difference (P > 0.05); there was no statistical difference of disease progression risk between the SPARC-positive and -negative groups (hazard ratio = 1.78, 95% confidence interval: 0.80–3.57, P > 0.05). However, SPARC-positive and -negative patients have shown a trend of DFS difference [Figure 2].
Figure 2: The disease-free survival curve for secreted protein acidic and rich in cysteine-positive and -negative patients

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 > Discussion Top


Breast cancer is the most common type of solid malignant tumors and its incidence rate continues to increase. With the continuous progress of molecular biology technology and the continuous development of targeting drugs, the prognosis and 5-year survival rate of breast cancer have significantly improved.[11],[12] Some patients, however, experience recurrence, and metastasis after surgery, recurrence, and metastasis have become important reasons for treatment failure. Therefore, exploring the molecular mechanism and the markers of breast cancer recurrence and metastasis is the key to improve the prognosis of breast cancer and the focus of breast cancer research.

The genes that encode SPARC proteins are located on the long arm of the human chromosome 5 at positions 31–33 and have a total length of 26.5 kb, including 10 exons and 9 introns.[13],[14] SPARC is a 40-kD single-stranded polypeptide that consists of cysteine-rich acidic glycoproteins. SPARC can be divided into four domains: A glutamic acid-rich region at the amino terminus near the calcium-binding domain, acysteine-rich domain, a hydrophilic domain, and an EF hand-shaped domain at the carboxyl terminal area.[14]

SPARC is a highly conserved ECM protein. It mainly functions to destroy cell adhesion,[15] regulate cell differentiation,[16] prevent cell dissemination, inhibit cell response to certain growth factors, regulate the ECM and the production of matrix metal protease, and impact the formation of new vessels.[17] Previous studies have shown that the SPARC protein is associated with the development and prognosis of various cancers, such as gastric and colorectal cancers.[18],[19],[20] Nevertheless, few research studies on the relationship between SPARC and the clinicopathological features and prognosis of breast cancer exist. The relationship between SPARC protein and the prognosis of breast cancer prognosis remains highly debated. Some researchers have suggested that the increased expression of SPARC protein in breast cancer epithelial cells is associated with a lower rate of survival.[21],[22] In addition, some studies have indicated that endogenous SPARC prevents the metastasis of MDA-MB-231 breast cancer cells by reducing the invasive activity of cells and the aggregation of blood platelets in tumor vessels; furthermore, the upregulation of SPARC could improve breast cancer prognosis.[23]

In this study, we found that a high proportion of patients with SPARC-positive breast cancer exhibited poorly differentiated tumors and were estrogen receptor-positive. In addition, the proportion of patients with lymph node metastases increased. These characteristics were associated with poor prognosis. We also divided the patients into positive and negative groups based on SPARC expression in breast cancer tissue. Furthermore, we constructed a survival curve using the risk proportional model to determine the relationship of SPARC expression with the postoperative recurrence or metastasis of tumors. We found that the 3-year progression-free survival rates of the SPARC-positive and SPARC-negative groups were not significantly different at 60.8% and 71.2%, respectively. Moreover, the risk of disease progression during the follow-up period between the two groups was not significantly different (P > 0.05). This result was consistent with those of Kim et al. In addition, SPARC-positive and SPARC-negative patients exhibited a tendency of difference in DFS. Therefore, the sample size should be increased and multicenter clinical research should be conducted to further clarify the role of SPARC in the development and progression of breast cancer and its feasibility as an indicator of breast cancer prognosis.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
 > References Top

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Motamed K, Blake DJ, Angello JC, Allen BL, Rapraeger AC, Hauschka SD, et al. Fibroblast growth factor receptor-1 mediates the inhibition of endothelial cell proliferation and the promotion of skeletal myoblast differentiation by SPARC: A role for protein kinase A. J Cell Biochem 2003;90:408-23.  Back to cited text no. 16
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Li Z, Li AD, Xu L, Bai DW, Hou KZ, Zheng HC, et al. SPARC expression in gastric cancer predicts poor prognosis: Results from a clinical cohort, pooled analysis and GSEA assay. Oncotarget 2016;7:70211-22.  Back to cited text no. 18
    
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Zhu A, Yuan P, Du F, Hong R, Ding X, Shi X, et al. SPARC overexpression in primary tumors correlates with disease recurrence and overall survival in patients with triple negative breast cancer. Oncotarget 2016;7:76628-34.  Back to cited text no. 21
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