Abstract
The aim of this investigation was to study the expression of caspases 3, 6 and 8 and their association to apoptosis in preneoplastic and neoplastic lesions of the breast. The material consisted of nine benign breast epithelial hyperplasias, 15 atypical hyperplasias, 74 in situ and 82 invasive carcinomas. The extent of apoptosis was assessed by the TUNEL method and caspase 3, 6 and 8 expression by immunohistochemistry with specific antibodies. Increased caspase 3 immunopositivity, as compared to staining of normal breast ductal epithelium, was seen in 22% of benign epithelial hyperplasias, 25% of atypical hyperplasias, 58% of in situ carcinomas and 90% of invasive carcinomas. The corresponding percentages for caspase 6 and 8 were 11%, 25%, 60%, 87% and 22%, 57%, 84%, 83% respectively. In high-grade in situ lesions there were significantly more cases with strong caspase 3, 6 and 8 immunoreactivity than in low- and intermediate-grade lesions (P = 0.0045, P = 0.049 and P = 0.0001 respectively). In invasive carcinomas, however, no association between a high tumour grade and caspase 3, 6 or 8 expression was found (P = 0.27, P = 0.26 and P = 0.69 respectively). The mean apoptotic index was 0.14 ± 0.14% in benign epithelial hyperplasias, 0.17 ± 0.12% in atypical hyperplasias, 0.61 ± 0.88% in in situ carcinomas and 0.94 ± 1.21% in invasive carcinomas. In all cases strong caspase 3, 6 and 8 positivity was significantly associated with the extent of apoptosis (P < 0.001, P = 0.015 and P = 0.050 respectively). The results show that synthesis of caspases 3, 6 and 8 is up-regulated in neoplastic breast epithelial cells in parallel to the increase in the apoptotic index and progression of the breast lesions.
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Alnemri, ES, Livingston, DJ, Nicholson, DW, Salvesen, G, Thornberry, NA, Wong, WW & Yuan, J (1996) Human ICE/CED-3 protease nomenclature. Cell 87: 171
Barge, RMY, Willemze, R, Vandenabeele, P, Fiers, W & Beyaert, R (1997) Differential involvement of caspases in apoptosis of myeloid leukemic cells induced by chemotherapy versus growth factor withdrawal. FEBS Lett 409: 207–210
Barnes, R & Masood, S (1990) Potential value of hormone receptor assay in carcinoma in-situ of breast. Am J Clin Pathol 94: 533–537
Brancolini, C, Lazarevic, D, Rodriguez, J & Schneider, C (1997) Dismantling cell–cell contacts during apoptosis is coupled to a caspase-dependent proteolytic cleavage of β-catenin. J Cell Biol 139: 759–771
Bur, ME, Zimarowski, MJ, Schnitt, SJ, Baker, S & Lew, R (1992) Estrogen receptor activity in carcinoma in-situ of the breast. Cancer 69: 1174–1181
Caulín, C, Salvesen, GS & Oshima, RG (1997) Caspase cleavage of keratin 18 and reorganization of intermediate filaments during epithelial cell apoptosis. J Cell Biol 138: 1379–1394
Chen, L, Marechal, V, Moreau, J, Levine, AJ & Chen, J (1997) Proteolytic cleavage of the mdm2 oncoprotein during apoptosis. J Biol Chem 272: 22966–22973
Chhanabhai, M, Krajewski, S, Krajewska, M, Wang, HG, Reed, JC & Gascoyne, RD (1997) Immunohistochemical analysis of interleukin-1 beta-converting enzyme/Ced-3 family protease, CCP32/Yama/Caspase-3 in Hodgkin’s disease. Blood 90: 2451–2455
Clem, RJ, Cheng, EH, Karp, CL, Kirch, DG, Ueno, K, Takahashi, A, Kastan, MB, Griffin, DE, Earnshaw, WC, Veliuona, MA & Hardwick, JM (1998) Modulation of cell death by Bcl-xL through caspase interaction. Proc Natl Acad Sci USA 95: 554–559
Droin, N, Dubrez, L, Eymin, B, Renvoize, C, Breard, J, Dimanche-Boitrel, MT & Solary, E (1998) Upregulation of CASP genes in human tumor cells undergoing etoposide-induced apoptosis. Oncogene 16: 2885–2894
Elston, CW & Ellis, IO (1991) Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long term follow-up. Histopathology 19: 403–410
Enari, M, Sakahira, H, Yokoyama, H, Okawa, K, Iwamatsu, A & Nagata, S (1998) A caspase-activated DNAase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391: 43–40
Faleiro, L, Kobayashi, R, Fearnhead, H & Lazebnik, Y (1997) Multiple species of CCP32 and Mch2 are the major active caspases present in apoptotic cells. EMBO J 16: 2271–2281
Fraser, A & Evan, G (1996) A license to kill. Cell 85: 781–784
Fujita, N & Tsuruo, T (1998) Involvement of bcl-2 cleavage in the acceleration of VP-16-induced U937 cell apoptosis. Biochem Biophys Res Commun 246: 484–488
Harvey, NL, Butt, AJ & Kumar, S (1997) Functional activation of Nedd2/ ICH-1 (Caspase-2) is an early process in apoptosis. J Biol Chem 272: 13134–13139
Helin, HJ, Helle, MJ, Kallioniemi, O-P & Isola, J (1989) Immunohistochemical determination of estrogen and progesterone receptors in human breast carcinoma. Cancer 63: 1761–1767
Holland, R, Hendricks, JHCL, Verbeck, ALM, Mravunac, M & Schuurmans-Stekhoven, JH (1990) Extent, distribution and mammographic/histological correlations of breast ductal carcinoma in situ. Lancet 335: 519–522
Janicke, RU, Sprengart, ML, Wati, MR & Porter, AG (1998) Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 273: 9357–9360
Kerr, JFR, Winterford, CM & Harmon, BV (1994) Apoptosis. Its significance in cancer and cancer therapy. Cancer 73: 2013–2026
Kluck, RM, Bossy-Wetzel, E, Green, DR & Nwemeyer, DD (1997) The release of cytochrome c from mitochondria: a primary site for bcl-2 regulation of apoptosis. Science 275: 1132–1136
Krajewska, M, Wang, H-G, Krajewski, S, Zapata, JM, Shabaik, A, Gascoyne, R & Reed, JC (1997) Immunohistochemical analysis of in vivo pattern of expression of CCP32 (caspase-3), a cell death protease. Cancer Res 57: 1605–1613
Krajewski, S, Krajewska, M, Shabaik, A, Miyashita, T, Wang, HG & Reed, JC (1994) Immunohistochemical determination of in vivo distribution of bax, a dominant inhibitor of bcl-2. Am J Pathol 145: 1323–133
Krajewski, S, Gascoyne, RD, Zapata, JM, Krajewska, M, Kitada, S, Chhanabhai, M, Horsman, D, Berean, K, Piro, LD, Fugier Vivier, I, Liu, YJ, Wang, HG & Reed, JC (1997) Immunolocalization of the ICE/Ced-3-family protease, CPP32 (caspase-3) in non-Hodgkin’s lymphomas, chronic lymphocytic leukemias, and reactive lymph nodes. Blood 89: 3817–3825
Kroemer, G (1997) The proto-oncogene bcl-2 and its role in regulating apoptosis. Nat Med 3: 614–620
Lipponen, P, Aaltomaa, S, Kosma, V-M & Syrjänen, K (1994) Apoptosis in breast cancer as related to histopathological characteristics and prognosis. Eur J Cancer 30A: 2068–2073
Liu, X, Naekyung, C, Yang, J, Jemmerson, R & Wang, J (1996) Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 86: 147–157
Mack, L, Kerkvliet, N, Doig, G & O’Malley, FP (1997) Relationship of a new histological categorization of ductal carcinoma in situ of the breast with size and the immunohistochemical expression of p53, c-erbB2, bcl-2 and Ki-67. Hum Pathol 28: 974–979
Manon, S, Chaudhuri, B & Guerin, M (1997) Release of cytochrome c and decrease of cytochrome c oxidase in bax-expressing yeast cells, and prevention of these effects by coexpression of bcl-xL. FEBS Lett 415: 29–32
Martins, LM, Kottke, T, Mesner, PW, Basi, GS, Sinha, S, Frigon, N, Tatar, E, Tung, JS, Bryant, K, Takahashi, A, Svingen, PA, Madden, BJ, McGormick, DJ, Dernshaw, WC & Kaufmann, SH (1997) Activation of multiple interleukin-1β converting enzyme homologues in cytosol and nuclei of HL-60 cells during etoposide-induced apoptosis. J Biol Chem 272: 7421–7430
Minn, AJ, Velez, P, Schendel, SL, Liang, H, Muchmore, SW, Fesik, SW, Fill, M & Thompson, CB (1997) Bcl-xL forms an ion channel in synthetic lipid membranes. Nature 385: 353–357
Mustonen, M, Raunio, H, Pääkkö, P & Soini, Y (1997) The extent of apoptosis is inversely associated with bcl-2 expression in premalignant and malignant breast lesions. Histopathology 31: 347–354
Muzio, M, Chinnaiyan, AM, Kischkel, FC, O’Rourke, K, Shevchenko, A, Ni, J, Scaffidi, C, Bretz, JD, Zhang, M, Gentz, R, Mann, M, Krammer, PH, Peter, ME & Dixit, VM (1996) FLICE, a novel FADD-homologous ICE/CED3-like protease is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 85: 817–827
Nagata, S (1997) Apoptosis by death factor. Cell 88: 355–365
Nijhawan, LP, Budihardjo, I, Srinivasula, SM, Ahmad, M, Alnemri, ES & Wang, X (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479–489
O’Malley, FP, Vnencak-Jones, CL, Dupont, WD, Parl, F, Manning, S & Page, DL (1994) p53 mutations are confined to the comedo type ductal carcinoma in situ of the breast. Immunohistochemical and sequencing data. Lab Invest 71: 67–72
Patel, T, Gores, GJ & Kaufmann, SH (1996) The role of proteases during apoptosis. FASEB J 10: 587–597
Rao, L, Perez, D & White, E (1996) Lamin proteolysis facilitates nuclear events during apoptosis. J Cell Biol 135: 1441–1455
Rosen, PP & Oberman, HA (1992). Tumors of the Mammary Gland. Atlas of Tumor Pathology. Third Series, Fascicle 7. Armed Forces Institute of Pathology: Washington, DC
Soini, Y, Virkajärvi, N, Lehto, V-P & Pääkkö, P (1996) Hepatocellular carcinomas with a high proliferation index and a low degree of apoptosis and necrosis are associated with a shortened survival. Br J Cancer 73: 1025–1030
Soini, Y, Pääkkö, P & Lehto, V-P (1998) Histopathological evaluation of apoptosis in cancer. Am J Pathol 153: 1041–1053
Tan, X, Martin, SJ, Green, DR & Wang, JYJ (1997) Degradation of retinoblastoma protein in tumor necrosis factor- and CD95-induced cell death. J Biol Chem 272: 9613–9616
Tavassoli, FA (1992). Pathology of the Breast. Appleton & Lange, Norwalk, Connecticut, USA. pp. 155–191
Thornberry, NA & Lazebnik, Y (1998) Caspases: enemies within. Science 281: 1312–1316
Thornberry, NA, Peterson, EP, Zhao, JJ, Howard, AD, Griffin, PR & Chapman, KT (1994) Inactivation of interleukin-1β converting enzyme by peptide (acyloxy)methyl ketones. Biochemistry 33: 3934–3940
Törmänen, U, Eerola, A-K, Rainio, P, Vähäkangas, K, Soini, Y, Sormunen, R, Bloigu, R, Lehto, V-P & Pääkkö, P (1995) Enhanced apoptosis predicts shortened survival in non-small cell lung carcinoma. Cancer Res 55: 5595–5602
van de Vijver, MJ, Peterse, JL, Mooi, WJ, Wisman, P, Lomans, J, Dalesion, O & Nusse, R (1988) Neu-protein overexpression in breast cancer. Association with comedo-type ductal carcinomas in situ and limited prognostic value in stage II breast cancer. New Engl J Med 319: 1239–1245
Virkajärvi, N, Pääkkö, P & Soini, Y (1998) Apoptotic index and apoptosis influencing proteins bcl-2, mcl-1, bax and caspases 3, 6 and 8 in pancreatic carcinoma. Histopathology 33: 432–439
Wang, TTY & Phang, JM (1995) Effects of estrogen on apoptotic pathways in human breast cancer cell line MCF-7. Cancer Res 35: 2487–2489
Yang, E & Korsmeyer, SK (1996) Molecular thanatopsis: a discourse on the bcl-2 family and cell death. Blood 88: 386–401
Yang, J, Liu, X, Bhalla, K, Kim, CN, Ibrado, AM, Cai, J, Peng, TI, Jones, DP & Wang, X (1997) Prevention of apoptosis by bcl-2; release of cytochrome c from mitochondria blocked. Science 275: 1129–1132
Author information
Authors and Affiliations
Additional information
Correspondence to:Y Soini
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Vakkala, M., Pääkkö, P. & Soini, Y. Expression of caspases 3, 6 and 8 is increased in parallel with apoptosis and histological aggressiveness of the breast lesion. Br J Cancer 81, 592–599 (1999). https://doi.org/10.1038/sj.bjc.6690735
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6690735
Keywords
This article is cited by
-
Arthrospira platensis-Mediated Green Biosynthesis of Silver Nano-particles as Breast Cancer Controlling Agent: In Vitro and In Vivo Safety Approaches
Applied Biochemistry and Biotechnology (2022)
-
Evaluation of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells Conditioning Medium (hWJ-MSCs-CM) or Scorpion Venom Breast Cancer Cell Line In Vitro
Journal of Gastrointestinal Cancer (2022)
-
Debris-stimulated tumor growth: a Pandora’s box?
Cancer and Metastasis Reviews (2021)
-
Role of Caspase-8 as a Prognostic Biomarker in Breast Cancer—A Pilot Study in Central India
Indian Journal of Gynecologic Oncology (2021)
-
Concomitance of downregulated active caspase-3 and upregulated X-chromosome linked inhibitor of apoptosis protein as a sensitive diagnostic approach for breast cancer
Molecular and Cellular Biochemistry (2019)