© Kamla-Raj 2006 Int J Hum Genet, 6(2): 133-138 (2006)
Protective Effect of 2-Deoxy-D-Glucose on Chemotherapeutic
Drugs Induced Damages on Peripheral Blood Lymphocytes
Exposed in-Vitro
P. Venkatachalam, V.R. Jayanth, Solomon F. D. Paul and V. Vettriselvi
Department of Human Genetics, Sri Ramachandra Medical College & Research Institute
(Deemed University), Porur, Chennai 600 116, Tamil Nadu, India
KEYWORDS Bleomycin; Mitomycin-C; chromosomal aberrations; sister chromatid exchanges
ABSTRACT The effect of 2-deoxy-D-glucose (2-DG), an antimetabolite of glucose was studied in peripheral blood
lymphocytes (PBL) exposed to radiomimetic drug bleomycin and an alkylating agent mitomycin-C. The PBL were
exposed to 2-DG (5 mM), 30 minutes pretreatment and with Bleomycin (10 to 80 μg/ml) and Mitomycin-C (2 to12
μg/ml) for three hours. The drug as well as 2-DG was removed by washing the cells with HBSS buffer. Then the cells
were cultured for 48 hours to study chromosomal aberrations (CA), Translocations (TL) and 72 hours for micronuclei
(MN) and Sister Chromatid Exchanges (SCE). Exposures of PBL to Bleomycin and Mitomycin-C showed, a
concentration dependent increase in the aberration frequencies, both in the presence and absence of 2-DG. While, the
regression analysis showed, that the presence of 2-DG reduced bleomycin induced TL, CA frequencies and Mitomycin-
C induced CA and MN frequencies significantly (P<0.001) when compared to PBL treated with the drugs alone,
Bleomycin induced MN frequencies and Mitomycin-C induced SCE’s reduction were not significant. The difference
could be attributed to the mechanism of the action of drugs on the cells. Furthermore the alteration in the cell cycle
kinetics, suggest that the presence of 2-DG during drug exposure, alter the cellular environment and delay the cell
proliferation and provide sufficient time to repair the damages, could resulted in the reduced aberration frequencies.
Corresponding Author: Dr P. Venkatachalam
Department of Human Genetics, Sri Ramachandra
Medical College & Research Institute (Deemed
University), Porur, Chennai 600 116, Tamil Nadu, India
Fax: 91+ 044-4767008, E-mail: venkip@yahoo.com
INTRODUCTION
It is well known that most of the radiation
used for therapy not only controls the division
of tumor cells but also mutagenic to normal cells
during multiple cycles of treatment (Bleehen et
al. 1974). Therefore, studies were reported on
the radio protective effect of compounds like 2-
Deoxy-D-gluocose (2-DG), Hoechst (Denison et
al. 1992) cysteine (Pattet et al. 1949), caffeine
(Franchitto et al. 1998) and calcium channel
blockers (Rajeeve and Kale 1995) on normal cells.
Of which the protective effect of 2-DG was
extensively studied in animal models (Jain et al.
1979) and in the lymphocytes of healthy
individuals (Kalia et al. 1988; Prabhu et al. 2004)
as well as glioma cells followed by radiation
(Dwarakanath et al. 1989). In addition to
radiotherapy, chemotherapy alone or in
combination with radiation plays an important
role to treat malignant diseases. The combined
use of chemotherapy and radiotherapy, in
patients may cause a degree of injury that would
be clinically toxic following either therapy alone
because, both may cause toxic to the same organ
by different mechanisms. Furthermore, the
response of cellular systems to therapeutic
agents depend on its effect on DNA, types of
lesions induced, capacity of the cells to repair
those lesions and cell cycle kinetics (Jain et al.,
1999). As our previous study had shown a
protective effect of 2-DG on radiation induced
chromosomal aberrations in the PBL (Prabhu et
al. 2004), the present study focus on the effect of
2-DG on radiomimectic drug bleomycin and
alkylating agent mitomycin-C induced damages
in PBL treated in-vitro. The PBL were exposed to
those drugs in the presence of 2-DG at G
0
phase
of the cells, which is the normal stage of majority
of the cells in the body at any given time, which
enables to some extent that the overall response
of tumors surrounding cell to therapeutic drugs.
The 5 mM concentration of 2-DG was selected
get an equimolar concentration of glucose in the
normal blood samples.
MATERIALS AND METHODS
In-vitro Treatment of Peripheral Blood
lymphocytes to Bleomycin and Mitomycin-C:
About 10 ml of peripheral blood was collected in
a heparinised sterile container from a healthy