115
Cancer Letters, 55 (1990) 115-120 Elsevier Scientific Publishers Ireland Ltd.
Chlorambucil
carcinogenesis
A. Cavalierea, N. Pietropaok, “Institute
Perugia
of
Pathological
University,
Anatomy
Perugia
in BALB/c
P.F. Alberti” and R. Vitalib and
Histology,
Division
Chlorambucil, a drug used in the treatment of neoplastic and non-neoplastic disease, was administered by gauage to BALB/c mice at a
dose of 1.0 mg/kg body wt. 5 times per week for 12 weeks to test its carcinogenicity. The survival was statistically reduced in treated animals of both sexes (P < 0.001). The treatment induced a significant increase in lung tumours (males, P < 0.001; females, P < 0.001) and lymphoreticular system tumours (males P < 0.01; females, P < 0.001) in both sexes and mammary carcinomas in female mice (P < 0.05). These results with other inuestigations reported in literature, suggest that chlorambucil is carcinogenic in laboratory animals, mutagenic and that it could be a potential carcinogenic hazard to man.
Keywords: chlorambucil; carcinogenesis
antineoplastic
drug;
Introduction Chlorambucil, an antineoplastic drug, has been used to treat patients with Hodgkin’s and Correspondence
to: Dr. Antonio
Cavaliere,
ogical Anatomy
and Histology,
Perugia
0304-3835/90/$03.50
0
Research
and
bDepartment
of Hygiene,
Institute of PatholUniversity,
Box 54,
non-Hodgkin’s lymphomas, chronic lymphocytic leukaemia, Kaposi’s disease and carcinoma of the breast, lung, testis and ovary. It is also used as an immunosuppressive agent in the treatment of systemic lupus erythematosus, rheumatoid arthritis, acute and chronic glomerulonephritis, nephrotic syndrome and Wegener’s granulomatosis. The drug is administered orally in doses of 0.1-0.2 mg/kg body wt. daily for at least 3-6 weeks. In the intermittent oral treatment doses of lo-20 mg daily are given for 2 weeks with rest periods of 2-4 weeks [3,24,26]. Side effects such as bone marrow hypoplasia, gastrointestinal disorders and hepatotoxicity are usually mild and rapidly-reversible though it is possible to induce serious bone marrow hypoplasia with high doses of the drug administered over long periods [3]. Long-term carcinogenesis tests done on mice and rats suggest that chlorambucil is carcinogenic. However in these experiments the chlorambucil was always administered intraperitoneally to the mice. This method is not used when administering the drug to humans. For this reason and since there is not data available for ascertaining the carcinogenicity of this drug on mice by oral administration and since this chemical is suspected of being carcinogenic in man [2,6,7] we decided to investigate its effect in BALB/c mice by oral administration.
1990 Elsevier Scientific Publishers Ireland Ltd.
Published and Printed in Ireland
Cancer
1990)
Summary
Perugia. Italy.
of
(Italy)
(Received 27 June 1990) (Revision received 18 September (Accepted 25 September 1990)
06100
mice
116
Materials and Methods
embedded in paraffin wax and routinely stained with haematoxilyn and eosin. Special stains were used only when necessary. The logrank test [lo] was used to analyse survival rates and the single tumour incidene trend calculated allowing for longevity [ 111.
Two groups of &week-old intact virgin male and female BALB/c/Cb/Se mice bred at the Division of Cancer Research, Institute of Pathological Anatomy, Perugia University, Italy, were used. Group 1: 53 males and 54 female mice received perorally by gavage 1.0 mg/kg body wt. chlorambucil in aqueous suspension 5 times per week for 12 weeks. Group 2: 50 male and 50 female control mice received only sterile saline solution. The chlorambucil (99.0% purity) was purchased from Sigma Chemical Co., St Louis, MO, U.S.A. and stored at - 20°C in the dark. All mice were housed 4-5 to a metal cage, maintained under identical environmental conditions and given water and commercial pellets (Laboratorio Dottori Piccioni. Brescia, Italy) ad libitum. During the experiment, both treated and control animals were inspected daily, killed if moribund and immediately autopsied. All tumours and pathologically altered organs were removed, fixed in 10% buffered formalin,
x of swvivom rm .-_ ____Lr_______, *. **.* . 1’. . . !, 90-
eo-
m-
A \ -
L-=.\ \,
Results
I and II give types, incidence of tumours and latency periods.
Lymphoreticular system
The rise in tumour incidence was statistically significant in both sexes (males P < 0.01; females P < 0.001). By the Pattengale and Taylor histological classification [9] the majority of tumours were follicular center ceil lymphomas and granulocytic leukaemias. Some
\\ , ‘?+,, .. . \ . ..
\
_-__--
ago
Cancer Letters, 55 (1990) 115-120 Elsevier Scientific Publishers Ireland Ltd.
Chlorambucil
carcinogenesis
A. Cavalierea, N. Pietropaok, “Institute
Perugia
of
Pathological
University,
Anatomy
Perugia
in BALB/c
P.F. Alberti” and R. Vitalib and
Histology,
Division
Chlorambucil, a drug used in the treatment of neoplastic and non-neoplastic disease, was administered by gauage to BALB/c mice at a
dose of 1.0 mg/kg body wt. 5 times per week for 12 weeks to test its carcinogenicity. The survival was statistically reduced in treated animals of both sexes (P < 0.001). The treatment induced a significant increase in lung tumours (males, P < 0.001; females, P < 0.001) and lymphoreticular system tumours (males P < 0.01; females, P < 0.001) in both sexes and mammary carcinomas in female mice (P < 0.05). These results with other inuestigations reported in literature, suggest that chlorambucil is carcinogenic in laboratory animals, mutagenic and that it could be a potential carcinogenic hazard to man.
Keywords: chlorambucil; carcinogenesis
antineoplastic
drug;
Introduction Chlorambucil, an antineoplastic drug, has been used to treat patients with Hodgkin’s and Correspondence
to: Dr. Antonio
Cavaliere,
ogical Anatomy
and Histology,
Perugia
0304-3835/90/$03.50
0
Research
and
bDepartment
of Hygiene,
Institute of PatholUniversity,
Box 54,
non-Hodgkin’s lymphomas, chronic lymphocytic leukaemia, Kaposi’s disease and carcinoma of the breast, lung, testis and ovary. It is also used as an immunosuppressive agent in the treatment of systemic lupus erythematosus, rheumatoid arthritis, acute and chronic glomerulonephritis, nephrotic syndrome and Wegener’s granulomatosis. The drug is administered orally in doses of 0.1-0.2 mg/kg body wt. daily for at least 3-6 weeks. In the intermittent oral treatment doses of lo-20 mg daily are given for 2 weeks with rest periods of 2-4 weeks [3,24,26]. Side effects such as bone marrow hypoplasia, gastrointestinal disorders and hepatotoxicity are usually mild and rapidly-reversible though it is possible to induce serious bone marrow hypoplasia with high doses of the drug administered over long periods [3]. Long-term carcinogenesis tests done on mice and rats suggest that chlorambucil is carcinogenic. However in these experiments the chlorambucil was always administered intraperitoneally to the mice. This method is not used when administering the drug to humans. For this reason and since there is not data available for ascertaining the carcinogenicity of this drug on mice by oral administration and since this chemical is suspected of being carcinogenic in man [2,6,7] we decided to investigate its effect in BALB/c mice by oral administration.
1990 Elsevier Scientific Publishers Ireland Ltd.
Published and Printed in Ireland
Cancer
1990)
Summary
Perugia. Italy.
of
(Italy)
(Received 27 June 1990) (Revision received 18 September (Accepted 25 September 1990)
06100
mice
116
Materials and Methods
embedded in paraffin wax and routinely stained with haematoxilyn and eosin. Special stains were used only when necessary. The logrank test [lo] was used to analyse survival rates and the single tumour incidene trend calculated allowing for longevity [ 111.
Two groups of &week-old intact virgin male and female BALB/c/Cb/Se mice bred at the Division of Cancer Research, Institute of Pathological Anatomy, Perugia University, Italy, were used. Group 1: 53 males and 54 female mice received perorally by gavage 1.0 mg/kg body wt. chlorambucil in aqueous suspension 5 times per week for 12 weeks. Group 2: 50 male and 50 female control mice received only sterile saline solution. The chlorambucil (99.0% purity) was purchased from Sigma Chemical Co., St Louis, MO, U.S.A. and stored at - 20°C in the dark. All mice were housed 4-5 to a metal cage, maintained under identical environmental conditions and given water and commercial pellets (Laboratorio Dottori Piccioni. Brescia, Italy) ad libitum. During the experiment, both treated and control animals were inspected daily, killed if moribund and immediately autopsied. All tumours and pathologically altered organs were removed, fixed in 10% buffered formalin,
x of swvivom rm .-_ ____Lr_______, *. **.* . 1’. . . !, 90-
eo-
m-
A \ -
L-=.\ \,
Results
I and II give types, incidence of tumours and latency periods.
Lymphoreticular system
The rise in tumour incidence was statistically significant in both sexes (males P < 0.01; females P < 0.001). By the Pattengale and Taylor histological classification [9] the majority of tumours were follicular center ceil lymphomas and granulocytic leukaemias. Some
\\ , ‘?+,, .. . \ . ..
\
_-__--
ago
