Table Of ContentRecent Results in Cancer Research
Fortschritte cler Krebsforschung
Progres clans les recherches sur Ie cancer
44
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Special Topics in Carcinogenesis
Edited by
E. Grundmann
With 54 Figures
Springer-Verlag Berlin· Heidelberg. New York 1974
Symposium of the "Gesellschaft zur Bekampfung der Krebskrankheiten
Nordrhein-Westfalen, e. V." DUsseldorf, 24th-25th March, 1972
Sponsored by the Swiss League against Cancer
ISBN-13: 978-3-642-80788-6 e-ISBN-13: 978-3-642-80786-2
DOl: 10.1007/978-3-642-80786-2
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CONTENTS
Welcome and Introduction. E. GRUNDMANN . . . . . . 1
General Problems of Carcinogenesis
Molecular Mechanisms in Chemical Carcinogenesis. P. N. MAGEE 2
Formation of Carcinogens From Precursors Occurring in the Environment:
New Aspects of Nitrosamine-induced Tumorgenesis. R. PREUSSMANN . 9
Intrauterine Induction of Cancer by the Experimental Application of Chemical
Substances. S. IVANKOVIC . . . . . . 16
The Replication of RNA Tumor Viruses. K. VON DER HELM. 21
Differentiation of Tumor Cells - Induction of Erythrocyte Membrane-Specific
Antigens in Friend Leukemia Cells. H. SUGANO, M. FURUSAWA, T. KAWA-
GUCHI, and Y. IKAWA . 30
Immunosuppressive Effect of Friend's Mouse Leukaemia and Anti-Thymocyte
Serum on Herpes Simplex Virus Infection. K. MUNK and H. RUNNEBAUM 45
Cytology and Histology of Cancer
Precancerous Lesions and their Detection and Diagnosis. 1. G. Koss 47
Preinvasive Carcinoma. H. HAMPERL 53
Enzyme Histochemistry of Human Tumors. Indications of Differentiation and
Functioning. R. G. J. WILLIGHAGEN andA. C. NIEUWENHUYZEN-KRUSEMAN. 57
Electron Microscopy of Oncocytomas and Carcinoid Tumors. H. SCHULZ 63
The Histology of Rare Human Tumors and their Relationships to General
Tumor Pathology. R. LAUMONIER and G. CHOMETTE • 69
Karyology of Carcinogenesis
Chromosome Changes in Human Malignant Tumors: An Evaluation. A. A.
SANDBERG . • . . . • . . • . . . . . . . . • . . . . 75
Chromosome Alteration in the Course of Serial Transplantions of Experimen-
tal Tumors and Aging of Tumor Stemline Cells. T. H. YOSIDA . 86
Early Changes in Ploidy and Cytogenetics of Liver Cells Mter Diethylnitros-
amine Treatment. H. WRBA, P. FISCHER, M. WEISSBERG, and 1. WIEST 94
VI Contents
Models of Experimental Carcinogenesis
Carcinogenesis in vitro. J. PONTEN . 98
Induction and Some Characteristics of "Minimal Deviation" and Other Trans-
plantable Rat Hepatomas. H. P. MORRIS and D. R. MERANZE . . " 103
Carcinogen-induced Cellular Thesaurismoses and Neoplastic Cell Transforma-
tion. P. BANNASCH 115
Experimental Intestinal Carcinogenesis and Polyp Development in Rats and
Mice. B. WIEBECKE . . . . 127
Tumor Induction in the Rat Kidney with Different Doses of DEN (Diethyl
nitrosamine): Frequency, Latency and Morphology of the Tumors. U. MOHR
and J. HILFRICH . . . . . . " 130
Respiratory Infections and the Pathogenesis of Lung Cancer. P. NETTESHEIM,
H. SCHREIBER, D. A. CREASIA, and C. B. RICHTER . 138
Experimental Tumors of the Nervous System. H. D. MENNEL . 158
Tumor Organotropy of N-nitrosomethylurea under the Influence of Hormonal
Contraceptives. C. THOMAS. 170
Role of Regional Lymph Nodes in Growth and Metastasis Formation of Me
thylcholanthrene-induced Sarcomas of Golden Hamsters. P. STRAULI and
R. LINDENMANN 174
Subject Index 183
List of Participants
BANNASCH, P., Dr., Pathologisches Institut der Universitat, D 8700 Wiirzburg, Luit
poldkrankenhaus
CHOMETTE, G., M.D., Service d'A natomie Pathologique, Groupe Hospitalier Pitie
Salpetriere, 83 Bd. de I'Hospital, F Paris 13 E
GEORGII, A., Dr., Pathologisches Institut der Medizinischen Hochschule, D 3000 Han
nover-Kleefeld, Roderbruchstr. 101
GRUNDMANN, E., Professor Dr., Pathologisches Institut der Universitat, D 4400 MUn
ster, Westring 17
HAMPERL, H., Dr., Pathologisches Institut der Universitat, D 5300 Bonn, Postfach
VON DER HELM, K., M.D., University of California, Department of Molecular Bio
logy, Berkeley, CA 94720jUSA
HILFRICH, J., Dr., Medizinische Hochschule Hannover, Abt. fUr experimentelle Patho
logie, Theoretische Institute I, D 3000 Hannover-Kleefeld, Roderbruchstr.l0l
IVANKOVIC, S., Dr., Deutsches Krebsforschungszentrum, Institut fUr experimentelle
Toxikologie und Chemotherapie, D 6900 Heidelberg, Kirschnerstr. 6
Koss, 1. G., M.D., Professor and Chairman, Department of Pathology, Albert Ein
stein College of Medicine at Montefiore Hospital and Medical Center, 111 East
210th Street, Bronx, NY 10467 jUSA
LAUMONIER, R., M.D., Faculte Mixte de Medicine et de Pharmacie, Hotel-Dieu, F 76
Rouen
MAGEE, P. N., M.D., Courtauld Institute of Biochemistry, The Middlesex Hospital,
London WIP 5PRjGreat Britain
MENNEL, H. D., Dr., Max-Planck-Institut fUr Hirnforschung, Abt. fUr Allgemeine
Neurologie, D 5000 Koln-Merheim, Ostmerheimer Str. 200
MOHR, U., Dr., Medizinische Hochschule Hannover, Abt. fUr experimentelle Patho
logie, D 3000 Hannover-Kleefeld, Roderbruchstr. 101
MORRIS, H. P., M.D., Howard University, Research Professor of Biochemistry, Can
cer Research Unit, Washington, D.C. 20001/USA
MUNK, K., Dr., Deutsches Krebsforschungszentrum, Direktor, Institut fUr Virusfor
schung, D 6900 Heidelberg, Kirschnerstr. 6
VIII List of Participants
NETTESHEIM, P., M.D., Oak Ridge National Laboratory, Post Office Box, Oak
Ridge, TN 37830/USA
PONTEN, J., M.D., Institut of Pathology, University, Uppsala/Schweden
PREUSSMANN, R., Dr., Deutsches Krebsforschungszentrum, Institut fUr experimentelle
Geschwulsterzeugung und -behandlung, D 6900 Heidelberg, Berliner Str. 27
SANDBERG, A. A., M.D., Chief of Medicine C, Roswell Park Memorial Institute De
partment of Health, 666 Elm Street, Buffalo, NY 14203/USA
SCHREIBER, H., M.D., Oak Ridge National Laboratory, Post office Box Y, Oak Ridge,
TN 37830/USA
SCHULZ, H., Dr., Chefarzt des Pathologischen Institutes, D 4500 OsnabrUck, Natru
per-T or-Wall 1
STRAULI, P., Dr., Institut fUr pathologische Anatomie der Universitiit, Abteilung
Krebsforschung, Kantonspital ZUrich, Schmelzbergstr. 12, CH 8006 Zurich
SUGANO, H., M.D., Cancer Institute, Toshima-ku, Tokyo/Japan
THOMAS, c., Dr., Pathologisches Institut der Universitiit, D 7800 Freiburg, Albert
straBe 19
WIEBECKE,B., Dr., Pathologisches Institut der Universitiit, D 8000 Munchen 15, Thal
kirchnerstr. 36
WILLIGHAGEN, R. G. J., M.D., Pathologisch Laboratorium der Rijksuniversiteit te
Leiden, Wassenaarseweg 62, Leiden/Holland
WOOD, S., M.D., Merck-Institute, Department of Experimental Pathology, Director,
Rahway, NJ 07065/USA
WRBA, H., Dr., Institut fUr Krebsforschung der Universitiit, A 1090 Wien, IX,
Borschkegasse 8 a
YOSIDA, T. H., National Institute of Genetics, Yata 1, Misima, Sizuoka-Ken/411
Japan
Welcome and Introduction
E.GRUNDMANN
On behalf of the "Gesellschaft zur Bekampfung der Krebskrankheiten Nordrhein
Westfalen e. V." I welcome you in Dusseldorf. During these two days problems of
oncogenesis have to be discussed. The participants have been selected with the inten
tion of giving a review of the various fields which have some bearing on this topic.
So you see here biochemists, cytologists, pathologists, immunologists, pharmacologists,
virologists, and so on. Only by extensive cooperation of all scientists concerned in
this field can this difficult problem of carcinogenesis ever be cleared up. Once the
causal relations have been identified, we may have a reliable basis for cancer treat
ment.
Research on oncogenesis is of great interest, but it is only a part of the worldwide
cancer campaign. It is our hope that this symposium will contribute towards helping
the cancer patient. He is the center of all our efforts.
Keeping this in mind - let us begin!
General Problems of Carcinogenesis
Molecular Mechanisms in Chemical Carcinogenesis
P. N.MAGEE
Covalent binding of metabolites of the hepatocarcinogenic azo dye p-dimethyl
aminoazobenzene (DAB) to proteins of the livers of rats to which it had been ad
ministered was observed by JAMES and ELIZABETH MILLER in 1947. Since then a
large amount of work on the interaction of many other chemical carcinogens with
cellular macromolecules has been reported. It is now well established that most,
perhaps all, chemical carcinogens become metabolically activated in the animal or
are themselves chemically reactive and that various of the activated forms interact
with tissue components in vivo. The terms precarcinogen, proximate carcinogen and
ultimate carcinogen have been introduced by the MILLERS to describe the original,
probably carcinogenic ally inactive compound, various intermediate metabolites of
the carcinogen with greater carcinogenic activity and the final active carcinogenic
form which is postulated to react at some crucial intracellular site and thus, in some
as yet unknown manner, cause a normal cell to become a malignant one. It ap
pears that, in general, the active forms of the chemical carcinogens are electrophilic
reactants which react with nucleophilic centres in the cell (MILLER, E. C. and MIL
LER, J. A., 1966; MILLER, J. A., 1970; MILLER, J. A. and MILLER, E. c., 1966, 1971).
Examples of such metabolic activations are the N-hydroxylation of aromatic amines
and azo dyes, followed by esterification (MILLER, 1970) and the reduction of 4-
nitroquinoline-N-oxide to 4-hydroxyaminoquinoline N-oxide (MATSUSHIMA and
SUGIMURA, 1971). It is now well established that the polycyclic hydrocarbons are
activated by microsomal hydroxylases in the liver and other tissues (NEBERT and
GELBOIN, 1969) but less is known concerning the nature of the active intermediates
although there is evidence that epoxide formation at the K region of molecules may
occur (GROVER, SIMS, HUBERMAN, MARQUARDT, KUROKI and HEIDELBERGER, 1971).
There is also evidence for microsomal metabolism of aflatoxins (SCHABORT and STEYN,
1969) and for binding of metabolites to cellular macromolecules (LI]INSKY, LEE and
GALLAGHER, 1970) but detailed information on the chemistry of these reactions is
lacking.
The N-dialkylnitrosamines undergo oxidative dealkylation in vivo to form al
kylating agents which react with cellular macromolecules (MAGEE and HULTIN, 1962;
MAGEE and FARBER, 1962) and the N-alkylnitrosamides appear to react in a similar
manner without enzymic activation (SWANN and MAGEE, 1968, 1971) although there
is evidence that in some cases the reaction is catalysed by sulphydryl compounds
(SCHOENTAL and RIVE, 1965; LAWLEY and THATCHER, 1970). The original suggestion
that the alkylating species were diazo alkanes is probably not correct in view of the
Molecular Mechanisms in Chemical Carcinogenesis 3
findings of LIJINSKY and his colleagues on the metabolism of fully deuterium-labelled
dimethyl- and diethylnitrosamines. These authors observed that alkylated cellular
components, in this case the 7-alkylguanines, isolated from livers of rats receiving
the deuterated carcinogens had molecular weights corresponding to transference of
the intact - CDa and - CD2CDa groups respectively rather than - CD2H and
- CDHCDa which would be formed if the diazoalkanes were obligatory interme
diates (LIJINSKY, Loo and Ross, 1968; Ross, KEEFER and LIJINSKY, 1971). This
evidence favours the intermediate formation of the alkyl diazonium cations
CHaCH2N2+ and CHaN2+ and/or the corresponding carbonium ions CHaCH2+
and CHa+ respectively. A similar reaction mechanism for N-methyl-N'-nitro-N
nitrosoguanidine is suggested by the work of LINGENS, HAERLIN and SUSSMUTH (1971)
with the deuterated compound. There is evidence suggesting a similar final metabolic
pathway for the naturally occurring carcinogen cycasin, CHa - N = N - CH 0-
+ 2
o
glucose. This compound is activated by the microbial flora of the gastrointestinal
tract, giving rise to methylazoxymethanol (MAM), CHa - N = N - CH 0H which
+ 2
o
is an alkylating agent in vitro (MATSUMOTO and HIGA, 1966) and in vivo (SHANK
and MAGEE, 1967; NAGATA and MATSUMOTO, 1969).
Several dialkylhydrazines, alkylazo and alkylazoxy compounds have been shown
to be potent carcinogens by DRUCKREY and his colleagues (DRUCKREY, 1970) who
suggested that this group of chemically related compounds might behave in a similar
manner in vivo to cycasin and MAM and give rise to the same ultimate carcinogenic
molecules (DRUCKREY, PREUSSMANN, IVANKOVIC, SCHMIDT, So and THOMAS, 1965).
Considerable recent interest has been shown in 1,2-dimethylhydrazine which induces
tumours of the colon in the rat (DRUCKREY, PREUSSMANN, MATSKIES and IVANKOVIC,
1967), the mouse (WIEBECKE, LOHRS, GIMMY and EDER, 1968) and the hamster (Oss
WALD and KRUGER, 1969). The metabolism of 1,2-dimethylhydrazine, 1,1-dimethyl
hydrazine and monomethylhydrazine has been studied in vivo and in vitro by REED
and his colleagues (DOST, REED and WANG, 1966; PROUGH, WITTKOP and REED,
1969, 1970; WITTKOP, PROUGH and REED, 1969) who have reported that these com
pounds undergo oxidative dealkylation by liver microsomal preparations in vitro.
Recent work in the laboratory of the author by Dr. A. HAWKS and Dr. P. F. SWANN
has shown that 1,2-dimethylhydrazine alkylates DNA and RNA of mouse liver
and colon in vivo (HAWKS, SWANN and MAGEE, 1972), thus providing experimental
evidence that this group of carcinogens does react with cellular macromolecules in
a manner similar to that of the nitroso compounds and the derivatives of methyl
azoxymethanol. The induction of tumours of the colon by 1,2-dimethylhydrazine
in the mouse has been confirmed and squamous carcinomas of the anus were also
found. In addition, administration of monomethylhydrazine to mice by repeated
subcutaneous injection has been in progress for about 32 weeks, but no colonic or
other tumour has so far been observed. In similar experiments with the 1,2-dimethyl
compound tumours of the colon were already present in several mice after this
period of administration so it appears that monomethylhydrazine may be consider
ably less potent as a colon carcinogen than the symmetrical dimethyl compound if,
indeed, it induces colonic tumours at all. Treatment of mice with [14C]monomethyl-
