Evaluation of high molecular weight cytokeratin (HMWCK), p27, C-X-C chemokine receptor type 4 (CXCR-4) and stromal cell-derived factor 1 (SDF-1) expressions related to tumor progression in breast cancer

Abstract

Objectives: Breast cancer is the most common cancer and the most common reason for cancer death in women population. The immunohistochemical markers which could have prognostic information are always needed.

Methods: This study included 365 cases of invasive ductal carcinoma (IDC), ductal carcinoma in situ (DCIS) and ductal epithelial hyperplasia. The cases divided into the following two groups according to the presence of cancer: 1) cancer group (298 cases; cases with IDC and DCIS), 2) non-cancer group (67 cases without cancer; cases with usual ductal epithelial hyperplasia [UDH] and atypical ductal epithelial hyperplasia [ADH]). All histological slides stained with high molecular weight cytokeratin (HMWCK), p27, C-X-C chemokine receptor type 4 (CXCR-4), stromal cell-derived factor 1 (SDF-1) immunohistochemically.

Results: IDC was present in 277 cases, of which 213 had pure IDC, and 64 had DCIS component adjacent to the invasive tumor. Twenty-one cases had only DCIS. Of 67 cases with epithelial hyperplasia, 31 had ADH, and 36 had UDH. Among cases with IDC, 143 had lymph node excision, of which 73 had metastasis in one or more lymph nodes, and 70 did not have metastatic disease. The expression of p27 was found to be significantly lower in the cancer group as compared to that in the non-cancer group (p < 0.0001). CXCR-4 expression in IDC was found to be higher than that of DCIS group. SDF-1 expression was observed to be significantly higher in cancer cases than that of non-cancer cases (p = 0.03).

Conclusions: The higher CXCR-4 and SDF-1 expressions are associated with tumor progression, tumor size, and lymph node status. In benign proliferative lesions, both HMWK and p27 expressions were helpful in differential diagnosis of borderline atypical ductal hyperplasia and DCIS. 

Keywords

• CXCR-4
• SDF-1
• p27
• HMWCK
• benign proliferative breast lesions
• ductal carcinoma in situ
• invasive ductal carcinoma
• tissue microarray

References

1. [1] Rosai J. Rosai and Ackerman's Surgical Pathology, Vol. 2, 9th ed., Mosby: London, 2004.
2. [2] Rosen PP. Rosen's Breast Pathology. Lippincott Williams&Wilking: Philadelphia, 2008.
3. [3] Mills SE. Sternberg's Diagnostic Surgical Pathology, Vol. 1, 4th ed., Lippincott Williams&Wilkins: Philadelphia, 2004.
4. [4] Dabbs D. Diagnostic Immunohistochemistry. Churchill Livingstone Elsevier: USA, 2006, 699-746 pp.
5. [5] Zhang JL, Zhang HY, Wei B, Lang ZQ, Bu H. [Role of cytokeratin expression in differential diagnosis of intraductal proliferative lesions of breast]. Zhonghua Bing Li Xue Za Zhi 2004;33:316-9. [Article in Chinese]
6. [6] Bonnefoy-Berard N, Aouacheria A, Verschelde C, Qemeneur L, Marçais A, Marvel J. Control of proliferation by Bcl-2 family members. Biochim Biophys Acta 2004;1644:159-68.
7. [7] Catzavelos C, Bhattacharya N, Ung YC, Wilson JA, Roncari L, Sandhu C, et al. Decreased levels of the cell-cycle inhibitor p27Kip1 protein: prognostic implications in primary breast cancer. Nat Med 1997;3: 227-30.
8. [8] Traub F, Mengel M, Luck HJ, Kreipe HH, von Wasielewski R. Prognostic impact of Skp2 and p27 in human breast cancer. Breast Cancer Res Treat 2006;99:185-91.

9. [9] Luster AD. Chemokines--chemotactic cytokines that mediate inflammation. N Engl J Med 1998;338:436-45.
10. [10] Hamada T, Tashiro K, Tada H, Inazawa J, Shirozu M, Shibahara K, et al. Isolation and characterization of a novel secretory protein, stromal cell-derived factor-2 (SDF-2) using the signal sequence trap method. Gene 1996;176: 211-4.
11. [11] Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity. Immunity 2000;12:121-7.
12. [12] Shirozu M, Nakano T, Inazawa J, Tashiro K, Tada H, Shinohara T, et al. Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF1) gene. Genomics 1995;28:495-500.
13. [13] Ma Q, Jones D, Borghesani PR, Segal RA, Nagasawa T, Kishimoto T, et al. Impaired B-lymphopoiesis, myelopoiesis, and derailed cerebellar neuron migration in CXCR4- and SDF-1-deficient mice. Proc Natl Acad Sci U S A 1998;95:9448-53.
14. [14] Wan WH, Fortuna MB, Furmanski P. A rapid and efficient method for testing immunohistochemical reactivity of monoclonal antibodies against multiple tissue samples simultaneously. J Immunol Methods 1987;103:121-9.
15. [15] Wu J, Shen ZZ, Lu JS, Jiang M, Han QX, Fontana JA, et al. Prognostic role of p27Kip1 and apoptosis in human breast cancer. Br J Cancer 1999;79:1572-8.
16. [16] Marlow R, Strickland P, Lee JS, Wu X, Pebenito M, Binnewies M, et al. SLITs suppress tumor growth in vivo by silencing Sdf1/Cxcr4 within breast epithelium. Cancer Res 2008;68:7819-27.
17. [17] Hwang ES, DeVries S, Chew KL, Moore DH 2nd, Kerliowske T, Thor A, et al. Patterns of chromosomal alterations in breast ductal carcinoma in situ. Clin Cancer Res 2004;10:5160-7.
18. [18] Sakorafas GH, Tsiotou AG. Ductal carcinoma in situ (DCIS) of the breast: evolving perspectives. Cancer Treat Rev 2000;26:103-25.
19. [19] Gal-Gombos EC, Esserman LE, Recine MA, Poppiti RJ, Jr. Large-needle core biopsy in atypical intraductal epithelial hyperplasia including immunohistochemical expression of high molecular weight cytokeratin: analysis of results of a single institution. Breast J 2002;8:269-74.
20. [20] Moinfar F, Man YG, Lininger RA, Bodian C, Tavassoli FA. Use of keratin 35betaE12 as an adjunct in the diagnosis of mammary intraepithelial neoplasia-ductal type--benign and malignant intraductal proliferations. Am J Surg Pathol 1999;23:1048-58.
21. [21] Tan P, Cady B, Wanner M, Worland P, Cukor B, Magi-Galluzzi C, et al. The cell cycle inhibitor p27 is an independent prognostic marker in small (T1a,b) invasive breast carcinomas. Cancer Res 1997;57:1259-63.
22. [22] Dorsam RT, Gutkind JS. G-protein-coupled receptors and cancer. Nat Rev Cancer 2007;7:79-94.
23. [23] Cabioglu N, Yazici MS, Arun B, Broglio KR, Hortobagyi GN, Price JE, et al. CCR7 and CXCR4 as novel biomarkers predicting axillary lymph node metastasis in T1 breast cancer. Clin Cancer Res 2005;11:5686-93.
24. [24] Yoon Y, Liang Z, Zhang X, Choe M, Zhu A, Cho HT, et al. CXC chemokine receptor-4 antagonist blocks both growth of primary tumor and metastasis of head and neck cancer in xenograft mouse models. Cancer Res 2007;67:7518-24.
25. [25] Chinni SR, Sivalogan S, Dong Z, Filho JC, Deng X, Bonfil RD, et al. CXCL12/CXCR4 signaling activates Akt-1 and MMP-9 expression in prostate cancer cells: the role of bone microenvironment-associated CXCL12. Prostate 2006;66:32-48.
26. [26] Wang J, Sun Y, Song W, Nor JE, Wang CY, Taichman RS. Diverse signaling pathways through the SDF-1/CXCR4 chemokine axis in prostate cancer cell lines leads to altered patterns of cytokine secretion and angiogenesis. Cell Signal 2005;17:1578-92.
27. [27] Schmid BC, Rudas M, Rezniczek GA, Leodolter S, Zeillinger R. CXCR4 is expressed in ductal carcinoma in situ of the breast and in atypical ductal hyperplasia. Breast Cancer Res Treat 2004;84:247-50.
28. [28] Silverstein MJ, Skinner KA, Lomis TJ. Predicting axillary nodal positivity in 2282 patients with breast carcinoma. World J Surg 2001;25:767-72.
29. [29] Liotta LA. An attractive force in metastasis. Nature 2001;410:24-5.
30. [30] Kang H, Mansel RE, Jiang WG. Genetic manipulation of stromal cell-derived factor-1 attests the pivotal role of the autocrine SDF-1-CXCR4 pathway in the aggressiveness of breast cancer cells. Int J Oncol 2005;26:1429-34.
31. [31] Kang H, Watkins G, Parr C, Douglas-Jones A, Mensel RE, Jiang WG. Stromal cell derived factor-1: its influence on invasiveness and migration of breast cancer cells in vitro, and its association with prognosis and survival in human breast cancer. Breast Cancer Res 2005;7: R402-10.
32. [32] Müller A, Homey B, Soto H, Ge N, Catron D, Buchanan ME, et al. Involvement of chemokine receptors in breast cancer metastasis. Nature 2001;410:50-6.
33. [33] Chen Y, Stamatoyannopoulos G, Song CZ. Down-regulation of CXCR4 by inducible small interfering RNA inhibits breast cancer cell invasion in vitro. Cancer Res 2003;63:4801-4.
34. [34] Woo SU, Bae JW, Kim CH, Lee JB, Koo BW. A significant correlation between nuclear CXCR4 expression and axillary lymph node metastasis in hormonal receptor negative breast cancer. Ann Surg Oncol 2008;15:281-5.