Original article
Application of -aminoisobutyric acid, L-methionine,
thymidine and 2-fluoro-2-deoxy-D-glucose to monitor effects
of chemotherapy in a human colon carcinoma cell line
Helmut Schaider 1, 2, Uwe Haberkorn 3, Martin R. Berger 2, Franz Oberdorfer 3, Iris Morr 3, Gerhard van Kaick 3
1 Department of Dermatology, KarI-Franzens University, Auenbruggerplatz 8, A-8036 Graz, Austria
2 Unit of Toxicology and Chemotherapy, German Cancer Research Center, Heidelberg, Germany
3 Department of Oncologic Diagnosis and Therapy, German Cancer Research Center, Heidelberg, Germany
Received 22 June and in revised form 16 September 1995
Abstract. Up to 4 h after treatment of human SW 707
colon carcinoma cells with the antineoplastic drug 4-
amino-N-(2'-aminophenyl)-benzamide (GOE 1734, di-
naline), the effects of tumour cell metabolism and prolif-
eration were examined in vitro. Four tracers which can
be labelled with isotopes suitable for positron emission
tomography (PET) were used for this purpose: (x-amino-
isobutyric acid (AIB) and methionine to study changes
in amino acid transport and protein synthesis, thymidine
to observe changes in tumour proliferation and 2-fluoro-
2-deoxy-D-glucose (FDG) to estimate glucose metabo-
lism. Dinaline showed an inhibition of the sodium-de-
pendent and -independent uptake of AIB. The methio-
nine uptake was found to increase shortly after therapy.
Thymidine incorporation into DNA was impaired and
the FDG uptake showed a maximally 2.2-fold enhance-
ment. Inhibition of AIB uptake suggests changes in ami-
no acid transport, whereas increased uptake of methio-
nine and FDG points to an enhancement of protein syn-
thesis and glycolysis caused by repair mechanisms. The
cytostatic and antiproliferative effect of dinaline, ob-
served in cell growth curves, could be demonstrated by
the impaired thymidine incorporation into DNA. This
study demonstrates that in vitro screening with radio-
tracers suitable for PET can help to clarify effects of
new antineoplastic substances on tumour cell metabo-
lism. These data may be applied to choose the appropri-
ate time schedule for monitoring therapeutic effects on
tumour tissue.
Key words: c~-Aminoisobutyric acid - L-Methionine -
Thymidine - 2-Fluoro-2-deoxy-D-(U-~4C)-glucose -
Dinaline
Eur J Nucl Med (1996) 23:55-60
Correspondence to: H. Schaider
Introduction
Currently used cytostatic drugs are able to achieve nor-
mal life spans only in patients with certain types of can-
cer. The responsive category does not include the most
frequent forms of malignant tumour, but it includes neo-
plasms with relatively short tumour volume doubling
times. Benzamides represent a group of pharmacologi-
cally active lipophilic compounds which were found to
affect poly-ADP-ribosylation [1], glucose metabolism,
DNA synthesis and cell viability [2]. 4-Amino-N(2'-ami-
nophenyl)-benzamide (dinaline, GOE 1734) is a com-
pound of this group and has been shown to be effective
against slowly growing, chemically induced rat mamma-
ry and colorectal carcinomas, osteosarcoma C22LR and
Brown Norway myeloid leukaemia [3-7]. A cytostatic
and cytotoxic effect of dinaline on various tumour cell
lines in vitro has been demonstrated, but the precise
mode of action has not yet been identified [8-10].
We have conducted a study to assess the effect of di-
naline on tumour cell metabolism, amino acid transport
and proliferation in vitro, c~-Aminoisobutyric acid
(AIB), methionine, thymidine and 2-fluoro-2-deoxy-D-
glucose (FDG) can be labelled with positron emitters
and have been shown to be applicable for in vivo mea-
surements [11-16]. Preclinical and clinical data support
the feasibility of using such radiotracers to validate the
response of cancers to chemotherapy [14-19]. To judge
the effects of new antineoplastic drugs on tumour cell
metabolism in vivo, in vitro studies with potential posi-
tron emission tomography (PET) tracers could be of pre-
dictive value for a better understanding of altered meta-
bolic processes accompanying chemotherapy.
Materials and methods
Chemicals. 4-Amino-N-(2'-aminophenyl)-benzamide was kindly
provided by G6deckeAG (Freiburg, Germany).3H-c~-aminoisobu-
tyric acid (methyl 3H) (329.3 GBq/mmol; 8.90 Ci/mmol) was ob-
tained from Du Pont, Nen Products (Boston, Mass.). 14C-L-me-
European Journal of Nuclear Medicine
Vol. 23, No. 1, January 1996 - © Springer-Verlag 1996