Expression of BCL2L12, a new member of apoptosis-related genes, in breast tumors

Maroulio Talieri; Eleftherios P. Diamandis; Nikos Katsaros; Dimitrios Gourgiotis; Andreas Scorilas

doi:10.1055/s-0037-1613411

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 89(06): 1081-1088
DOI: 10.1055/s-0037-1613411

Cellular Proteolysis and Oncology

Schattauer GmbH

Expression of BCL2L12, a new member of apoptosis-related genes, in breast tumors

Authors

Maroulio Talieri

¹“G. Papanicolaou” Research Center of Oncology “Saint Savas” Hospital Athens, Greece
Eleftherios P. Diamandis

²Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, and Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada
Nikos Katsaros

³National Center for Scientific Research “Demokritos”, IPC, Athens Greece
Dimitrios Gourgiotis

⁴Research Laboratories, Second Department of Pediatrics, School of Medicine, University of Athens, Athens, Greece
Andreas Scorilas

³National Center for Scientific Research “Demokritos”, IPC, Athens Greece

Abstract

Summary

Apoptosis, a normal physiological form of cell death, is critically involved in the regulation of cellular homeostasis. If the delicate balance between cell death and cell proliferation is altered by a defect in the normal regulation of apoptosis signaling, a cell population is able to survive and accumulate, thereby favoring the acquisition of further genetic alterations and promoting tumorigenesis. Dysregulation of programmed cell death mechanisms plays an important role in the pathogenesis and progression of breast cancer, as well as in the responses of tumors to therapeutic intervention. Overexpression of anti-apoptotic members of the Bcl-2 family such as Bcl-2 and Bcl-XL has been implicated in cancer chemoresistance, whereas high levels of pro-apoptotic proteins such as Bax promote apoptosis and sensitize tumor cells to various anticancer therapies. Recently, a new member of the Bcl-2 family, BCL2L12, was cloned. The BCL2L12 gene is constitutively expressed in many tissues, suggesting that the encoded protein serves an important function in different cell types. In the present study, the expression of BCL2L12 gene was analyzed by reverse transcription-PCR (PT-PCR) in 70 breast cancer tissues. Our results indicate that BCL2L12 positive breast tumors are mainly of lower stage (I/II) or grade (I/II) (p=0.02 or p=0.04 respectively). Cox regression analysis revealed that BCL2L12 expression is positively related to disease-free (DFS) and overall survival (OS) at both univariate and multivariate analysis (p=0.021, p=0.029, p=0.032, p=0.044 respectively). Kaplan-Meier survival curves also demonstrated that patients with BCL2L12-positive tumors have significantly longer DFS and OS (p=0.002 and p<0.001 respectively). BCL2L12 expression may be regarded as a new independent favorable prognostic marker for breast cancer.

Part of this paper was originally presented at the joint meetings of the 16th International Congress of the International Society of Fibrinolysis and Proteolysis (ISFP) and the 17th International Fibrinogen Workshop of the International Fibrinogen Research Society (IFRS) held in Munich, Germany, September, 2002.

Keywords

Apoptosis - BCL2L12 - Bcl2 gene family - breast cancer - prognostic markers

Full Text

References

References
1 Searle J, Kerr JFR, Bishop CJ. Necrosis and apoptosis: distinct modes of cell death with fundamentally different significance. Pathol Annu 1982; 17: 229-59.
2 Buja LM, Eigenbrodt ML, Eigenbrodt EH. Apoptosis and necrosis. Basic types of cell death. Arch Pathol Lab Med 1993; 117: 1208-14.
3 Reed JC. Mechanisms of apoptosis. Warner-Lambert/Park-Davis Award Lecture. Am J Pathol 2000; 157: 1415-30.
4 Zimmermann KC, Bonzon Ch, Green DR. The machinery of programmed cell death. Pharmacol Ther 2001; 92: 57-70.
5 Wyllie AH. Apoptosis and the regulation of cell numbers in normal and neoplastic tissues: an overview. Cancer Metastasis Rev 1992; 11: 95-103.
6 Choi Ws, Yoon SY, Chang II. et al. Correlation between structure of Bcl-2 and its inhibitory function of JNK and caspase activity in dopaminergic neuronal apoptosis. J Neuro-chem 2000; 74: 1621-6.
7 Zha H, Aime-Sempe C, Sato T, Reed JC. Proapoptotic protein Bax heterodimerizes with Bcl-2 and homodimerizes with Bax via a novel domain (BH3) distinct from BH1 and BH2. J Biol Chem 1996; 271: 7440-4.
8 Reed JC. Dysregulation of apoptosis in cancer. J Clin Oncol 1999; 17: 2941-53.
9 Tamm I, Schriever F, Dorken B. Apoptosis implications of basic research for clinical oncology. Lancet Oncol 2001; 2: 33-42.
10 Hickman JA. Apoptosis induced by anticancer drugs. Cancer Metastasis Rev 1992; 11: 121-39.
11 Verheij M, Batrelink H. Radiation-induced apoptosis. Cell Tissue Res 2000; 301: 133-42.
12 Adams JM, Cory S. The Bcl-2 protein family: Arbiters of cell survival. Science 1998; 281: 1322-6.
13 Reed JC, Zha H, Aime-Sempe C. et al. Structure-function analysis of Bcl-2 family proteins. Regulators of programmed cell death. Adv Exp Med Biol 1996; 406: 99-112.
14 Korsmeyer SJ. BCL-2 gene family and the regulation of programmed cell death. Cancer Res 1999; 59: 1693S-700S..
15 Bar-Sagi D, Rotin D, Batzer A. et al. SH3 domain direct cellular localization of signaling molecules. Cell 1993; 74: 83-91.
16 Wang B, Zou JX, Ek-Rylander B. et al. R-Ras contains a proline-rich site that binds to SH3 domains and is required for integrin activation by R-Ras. J Biol Chem 2000; 275: 5222-7.
17 Alexandropoulos K, Cheng G, Baltimore D. Proline-rich sequences that bind to Src homology 3 domains with individual specificities. Proc Natl Acad Sci U S A 1995; 92: 3110-14.
18 Scorilas A, Kyriakopoulou L, Yousef G. et al. Molecular cloning, physical mapping and expression analysis of a novel gene, BCL2L12, encoding a proline-rich protein with a highly conserved BH3 domain of the Bcl-2 family. Genomics 2001; 72: 217-21.
19 Scorilas A, Yousef G, Diamandis EP. Identification and characterization of a novel human testis specific kinase substrate gene which is down-regulated in testicular carcinomas. Biochem Biophys Res Commun 2001; 285: 400-8.
20 Scorilas A, Kyriakopoulou L, Katsaros D. et al. Cloning of a gene (SR-A1), encoding for a new member of the human Ser/Arg-rich family of Pre-mRNA splicing factors: Over-expression in aggressive ovarian cancer. Br J Cancer 2001; 85: 190-8.
21 Scorilas A, Levesque AL, Ashworth KL. et al. Cloning, structural characterization, physical mapping and expression pattern of the human alpha-adaptin A gene. Gene 2001; 289: 191-9.
22 Scorilas A, Black MH, Talieri M. et al. Genomic organization of the human protein argi-nine methyltransferase 1 gene (PRMT1): Differential expression of splice variants in breast cancer Biochem Biophys Res Commun. 2000; 278: 349-259.
23 Scott-Conor CE, Christie DW. Cancer staging using the American Joint Committee on Cancer TNM System. J Am Coll Surg 1995; 181: 182-8.
24 Doussal VL, Tubiana-Hulin M, Friedman S. et al. Prognostic value of histilogigic grade nuclear components of Scarff-Bloom-Richardson (SBR): an improved score modification based on a multivariate analysis of 1262 invasive ductal breast carcinomas. Cancer 1989; 64: 1914-22.
25 Dressler LG, Seamer LC, Owens MA. et al. DNA flow cytometry and prognostic factors in 1131 frozen breast cancer specimens. Cancer 1988; 61: 420-7.
26 Scorilas A, Talieri M, Ardavanis A. et al. Polyadenylate polymerase enzymatic activity in mammary tumor cytosols: A new independent prognostic marker in primary breast cancer. Cancer Res 2000; 60: 5427-33.
27 Scatchard G. The attraction of proteins for small molecules and ions. Ann NY Acad Sci 1949; 51: 660-72.
28 Alexiera-Figusch J, VAN Putten WLS, Blankestein MA. et al. The prognostic value and relationships of patient characteristics, estrogen and progestin receptors and site of relapse in primary breast cancer. Cancer 1988; 61: 758-68.
29 Reiher A, Kolb R, Reiner G. et al. Prognostic significance of steroid hormone receptor and histopathological characterization of human breast cancer. J Cancer Res Clin Oncol 1987; 113: 285-90.
30 Cox DR. Regression models and life tables R. Stat Soc. B, 1972; 34: 187-202.
31 Kaplan EL, Meier P. Nonparametric estimation from incomplete observations. J Am Stat Assoc 1958; 53: 457-81.
32 Sjostrom J, Blomqvist C, von Boguslawski K. et al. The predictive value of bcl-2, bax, bcl-Xl, bag-1, fas, and fasL for chemotherapy response in advanced breast cancer. Clin Cancer Res 2002; 8: 811-6.
33 Silvestrini R, Benini E, Verneroni S. et al. p53 and bcl-2 expression correlates with clinical outcome in a series of node-positive breast cancer patients. J Clin Oncol 1996; 14: 1604-10.
34 Zhang G, Kimijina I, Abe R. et al. Apoptotic index correlated to Bcl-2 and p53 protein expression, histological grade and prognosis in invasive breast cancers. Anticancer Res 1998; 18: 1989-98.
35 Bukholm I, Bukhlm G, Nesland J. Reduced expression of both Bax and Bcl-2 is independently associated with lymph node metastasis in human breast carcinomas. APMIS 2002; 110: 214-20.
36 Krajewski S, Krajewska M, Turner BC. et al. Prognostic significance of apoptosis regulators in breast cancer. Endocrine-Related Cancer 1999; 6: 29-40.
37 Simonian PL, Grillot DA, Andrews DW. et al. Bax homodimerization is not required for Bax to accelerate chemotherapy-induced cell death. J Biol Chem 1996; 271: 32073-77.
38 Ottilie S, Diaz JL, Chang J. et al. Structural and functional complementation of an inactive Bcl-2 mutant by Bax truncation. J Biol Chem 1997; 272: 16955-61.
39 Akahani S, Nangia-Makker P, Inohara H. et al. Galectin-3: a novel antiapoptotic molecule with a functional BH1 (NWGR) domain of Bcl-2 family. Cancer Res 1997; 57: 5272-6.
40 Baliga BC, Kumar S. Role of BCL-2 family of proteins in malignancy. Hematol Oncol 2002; 20: 63-74.