Abstract
Testis tumors of embryonal origin (ten metastasized, six non-metastasized) and 17 mixed testis cell carcinomas (eight metastasized, nine non-metastasized) were examined. A triple immunofluorescence microscopic labeling procedure allowed the simultaneous detection of two features of apoptosis, namely morphological changes in the nucleus (DNA condensation visualized by DAPI staining) and the process of DNA fragmentation (TdT-assay) in tumor cells as well as T-cells (recognized by their CD45RO epitope). Both methods for apoptosis detection showed similar apoptotic indices (AI) only in 2.6% of all tumors. Most tumors (81.6%) showed more cells with DNA fragments than condensed chromatin, but in a number of cases (10.5%) the opposite pattern was found. These data add to the few published in vivo examinations of apoptosis using different methods and help to explain differences in the judgment of apoptosis significance for tumor prognosis. With regard to tumorigenesis, non-metastasized testis tumors were characterized by higher AIs of tumor cells and T-cells compared with metastasized tumors, which could be interpreted as a characteristic of tumors in an earlier stage of their development into an apoptosis-resistant phenotype. For the first time, in metastasized tumors a 5 to 25-fold increase of the T-cell’s AIs over the corresponding AIs of tumor cells was shown. This suggests a successful counterattack of tumor cells, thus supporting the process of metastasis. However, only ten out of 33 tumors revealed these AI changes, which again highlights that tumor biology cannot be predicted by a single parametric approach. It remains to be seen whether these characteristics might be suitable for a reliable prediction of metastasis.
Similar content being viewed by others
References
Abend M, Schmelz HU, Kraft K, Rhein AP, van Beuningen D, Sparwasser, C (1998) Inter-comparison of apoptosis morphology with active DNA cleavage on single cells in vitro and on testis tumours. J Pathol 185:419
Aftabuddin M, Yamadori I, Yoshino T, Kondo E, Akagi, T (1995) Correlation between the number of apoptotic cells and expression of the apoptosis-related antigens Fas, Leyand bcl-2 protein in non-Hodgkins lymphomas. Pathol Int 45:422
Agui T, McConkey DJ, Tanigawa N (2002) Comparative study of various biological parameters, including expression of surviving, between primary and metastatic human colonic adenocarcinomas. Anticancer Res 22:1769
Brown JM, Wouters BG (1999) Does apoptosis contribute to tumor cell sensitivity to anticancer agents? In: Hickman JA, Dive C (eds) Apoptosis and cancer chemotherapy. Humana Press, Totowa, NJ, pp. 1–18
Chen YL, Wang JY, Chen SH, Yang BC (2002) Granulocytes mediates the Fas-L-associated apoptosis during lung metastasis of melanoma that determines the metastatic behaviour. Br J Cancer 87:359
Del Bino G, Darzynkiewicz Z, Degraef C, Mosselmans R, Fokan D, Galand P (1999) Comparison of methods based on annexin-V binding, DNA content or TUNEL for evaluating cell death in HL-60 and adherent MCF-7 cells. Cell Prolif 32:25
El-Ahmady O, El-Salahy E, Mahmoud M, Wahab MA, Eissa S, Khalifa A (2002) Multivariate analysis of bcl-2, apoptosis, P53 and HER-2/neu in breast cancer: a short-term follow-up. Anticancer Res 22:2493
Fernandez Y, Gu B, Martinez A, Torregrosa A, Sierra A (2002) Inhibition of apoptosis in human breast cancer cells: role in tumor progression to the metastatic state. Int J Cancer 101:317
Fisher, DE (1994) Apoptosis in cancer therapy: crossing the threshold. Cell 78:539
Izawa JI, Sweeney P, Perrotte P, Kedar D, Dong Z, Slaton JW, Karashima T, Inoue K, Benedict WF, Dinney CP (2002) Inhibition of tumorigenicity and metastasis of human bladder cancer growing in athymic mice by interferon-beta gene therapy results partially from various antiangiogenic effects including endothelial cell apoptosis. Clin Cancer Res 8:1258
Kerr JFR, Wyllie AH, Currie AR (1972) Apoptosis: a basis biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239
Kradin RL, Bhan AK (1993) Tumor infiltrating lymphocytes. Lab Invest 69:635
Levine EL, Davidson SE, Roberts SA, Chadwick CA, Potten CS, West CML (1994) Apoptosis as predictor of response to radiotherapy in cervical carcinoma. Lancet 44:344
Li L, Yan L, Wei Y, Liu Z, Wang Z, Lei S, Huang G (1999) Significance of apoptosis status and apoptosis-associated antigen expression in human colorectal adenocarcinoma sequence. Hua Xi Yi Ke Da Xue Xue Bao 30:185
Lopez T, Hanahan D (2002) Elevated levels of IGF-1 receptor convey invasive and metastatic capability in a mouse model of pancreatic islet tumorigenesis. Cancer Cell 1:339
Lu QL, Elia G, Lucas S, Thomas JA (1999) Bcl-2 proto-oncogene expression in Epstein-Barr-virus-associated nasopharyngeal carcinoma. Int J Cancer 53:29
Oberhammer F, Wilson JW, Dive C, Morris ID, Hickman JA, Wakeling AE, et al. (1993) Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J 12:3679
Oliver L, Cordel S, Barbieux I, LeCabellec MT, Meflah K, Gregoire M, Vallette FM (2002) Resistance to apoptosis is increased during metastatic dissemination of colon cancer. Clin Exp Metast 19:175
Richter EN, Oevermann K, Buentig N, Storkel S, Dallmann I, Atzpodien J (2002) Primary apoptosis as a prognostic index for the classification of metastatic renal cell carcinoma. J Urol 168:460
Rupnow BA, Knox SJ (1999) The role of radiation-induced apoptosis as a determinant of tumor responses to radiation therapy. Apoptosis 4:115
Scaife CL, Kuang J, Wills JC, Trowbridge DB, Gray P, Manning BM, Eichwald EJ, Daynes RA, Kuwada SK (2002) Nuclear factor kappaB inhibitors induce adhesion-dependent colon cancer apoptosis: implications for metastasis. Cancer Res 62:6870
Schmelz HU, Abend M, Kraft K, van Beuningen D, Pust R, Sparwasser C (1999) Apoptosis in human embryonal cell carcinoma; preliminary results. Urol Res 27:368
Schmelz HU, Abend M, Kraft K, Hauck EW, Weidner W, van Beuningen D, Sparwasser, C (2002) Fas/Fas ligand system and apoptosis induction in testicular carcinoma. Cancer 95:73
Schmitt CA, Lowe SW (1999) Apoptosis and therapy. J Pathol 187:127
Shin MS, Kim HS, Lee SH, Lee JW, Song YH, Kim YS, Park WS, Kim SY, Lee SN, Park JY, Lee JH, Xiao W, Jo KH, Wang YP, Lee KY, Park YG, Kim SH, Lee JY, Yoo NJ (2002) Alterations of Fas-pathway genes associated with nodal metastasis in non-small cell lung cancer. Oncogene 21:4129
Staunton MJ, Gaffney EF (1995) Tumor type is a determinant of susceptibility to apoptosis. Am J Clin Pathol 103:300
Steel G (2001) The case against apoptosis (review article). Acta Oncol 40:968
Termuhlen PM, Sweeney-Gotsch BM, Berman RS, Ellis LM, Bucana C, Shen Y, Cleary KR, McConkey DJ (2002) Increased apoptosis in metastatic human colonic adenocarcinomas Cancer Biol Ther 1:58
Walker PR, Sikorska M (1997) New aspects of the mechanism of DNA fragmentation in apoptosis. Biochem Cell Biol 75:287
Walker PR, Pandey S, Sikorska M (1995) Degradation of chromatin in apoptotic cells. Cell Death Diff 2:97
Wheeler JA, Stephens LC, Tornos C, et al (1995) ASTRO research fellowship: apoptosis as a predictor of tumor response to radiation in stage IB cervival carcinoma. Int J Rad Oncol Biol Phys 32:1487
Yamamoto T, Oda K, Kubota T, Miyazaki K, Takenouti Y, Nimura Y, Hamaguchi M, Matsuda S (2002) Expression of p73 gene, cell proliferation and apoptosis in breast cancer: Immunohistochemical and clinicopathological study. Oncol Rep 9:729
Zhu Q, Deng C (2002) Apoptosis of lymphocytes induced by Fas/FasL of colon cancer cells. Ai Zheng 21:272
Grobholz R, Zentgraf H, Kohrmann KU, Bleyl U (2002) Bax, Bcl-2, fas and Fas-L antigen expression in human seminoma: correlation with the apoptotic index. APMIS 110:724
Acknowledgments
We very much appreciate the accurate technical work of A. Pflüger. We also wish to stress our thanks to Mrs. B. Hay, Department of Biometry and Medical Documentation, University of Ulm, who contributed valuable recommendations from a biomathematical point of view. This work was supported by a special research grant from the German Ministry of Defense.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Abend, M., Port, M., Schmelz, H.U. et al. Significance of apoptosis in metastasizing testis tumors. Urol Res 32, 28–35 (2004). https://doi.org/10.1007/s00240-003-0370-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00240-003-0370-x