Evaluation and characterization of micronuclei induced by the antitumour agent ASE [3beta-hydroxy-13alpha-amino-13,17-seco-5alpha-androstan-17-oic-13,17-lactam-p-bis(2-chloroethyl)amino phenylacetate] in human lymphocyte cultures

Abstract

increased lifespan over controls in the treatment of L1210 N.A.Demopoulos leukemia in mice and a 150% increased lifespan in the treatment of P388 leukemia (Wampler and Catsoulacos, 1977).Division of Genetics, Cell and Developmental Biology, Department of Moreover, ASE has given good activity against B 16 melanoma Biology, University of Patras, 26500 Patras, Greece in C 57 b 1 mice and T 8 Guerrin tumors in rats (Catsoulacos and 3β-Hydroxy-13α-amino-13,17-seco-5α-androstan-17- Wampler, 1982). In addition, ASE alone or in combination oic-13,17-lactam-p-bis(2-chloroethyl)amino phenylacetate with a different modified steroid, the homo-aza-steroidal ester (ASE) is a homo-aza-steroidal ester of p-bis(2-chloroethyl)of o-bis(2-chloroethyl)aminobenzoic acid, proved to exert amino phenyl acetic acid and has been shown to display enhanced antitumor activity in a synergistic manner antineoplastic, mutagenic and genotoxic activity. In the (Papageorgiou et al., 1999). It is noteworthy that although present study an effort has been made to evaluate the ASE exerts about the same antineoplastic activity as ability of ASE to induce micronuclei (MN) in human chlorambucil, it is less toxic with a more prolonged action lymphocytes treated in vitro using the cytokinesis-block (Stevenson and Patel, 1973; Catsoulacos et al., 1978). assay. Lympocytes were treated with different concentraAdditionally, ASE has shown mutagenic properties. It was tions of ASE (0.1, 0.25, 0.5, 1, 2.5, 5, 10 and 20 µg/ml) at found to be positive in strains TA1535 and TA100 as well as two different cell culture times, 21 and 41 h after culture in strain TA102 with and without metabolic activation in the initiation. ASE treatment lasted until cell harvest, for Salmonella/microsome system (Athanasiou and Arzimanoglou, 51 and 31 h, respectively. Two types of cultures were used, 1986). The compound has also been shown to induce sister whole blood and isolated lymphocyte cultures. The content chromatid exchange in human lymphocytes (Mourelatos et al., of induced MN was identified by FISH analysis, using an 1987) as well as in CHO cells (Athanasiou et al., 1983). ASE α-satellite DNA probe, in binucleate cells. Our results was also determined to decrease protein synthesis rate in suggest that ASE is capable of increasing MN frequencies ovaries of Drosophila melanogaster (Stephanou et al., 1991). in human lymphocytes under both culture conditions. ThisFinally, ASE was identified as displaying genotoxic, cytostatic increase is related to the concentration in a linear doseand antineoplastic properties (Nikolaropoulos et al., 1997). dependent manner and is also dependent on the duration Human lymphocytes cultured in vitro have been widely of treatment. FISH analysis has shown that the induced used as an appropriate system to identify the genetic activity MN resulted mainly from breakage events. Additionally, of various chemical compounds. They are a more or less a weak aneugenic effect was found at the higher conhomogeneous cell population, being all in the G 0 phase, an...

Genotoxicity of hydrochlorothiazide in cultured human lymphocytes. I. Evaluation of chromosome delay and chromosome breakage

“…However, it has been noted previously that there are very few genotoxic agents that induce only C þ MN or only C À MN [Schuler et al, 1997]. Indeed, many clastogens [Vlachodimitropoulos et al, 1997;Andrianopoulos et al, 2000;Kouloumenta et al, 2005] and aneugens [Bakou et al, 2002] produce both types of MN. Myhr et al [1990] reported that …”

Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond

“…MN caused by breakage events were noticed in human lymphocytes treated with antineoplastic drug ASE (Andrianopoulos et al, 2000). Teniposide, an anti-tumor drug used for treatment of childhood acute lymphocytic leukemia, induced MN with a peak frequency at 16 h after treatment, which was correlated with cell survival decline (Adiga and Jagetia, 1999).…”

Aneugenic potential of the anticancer drugs melphalan and chlorambucil. The involvement of apoptosis and chromosome segregation regulating proteins

“…It was reported that melphalan, in addition to its clastogenicity, increased micronuclei (MN) originating from chromosome delay events in MCL-5 and h1A1v2 cells (Phousongphouang et al, 2000). Although ASE, a homo-aza-steroidal ester of PHE [p-N,N-bis(2-chloroethyl) aminophenylacetic acid], induced MN that primarily formed owing to chromosome breakage, a weak aneugenic potential was reported in human lymphocyte cultures in vitro (Andrianopoulos et al, 2000). Kouloumenta et al (2005) showed that two steroidal analogs of PHE exerted dual genetic activity, clastogenic and aneugenic.…”