In vitro micronucleus bioassay of human peripheral lymphocytes for adriamycin in the presence of cyclophosphamide and urines of patients administered anticancer drugs

Abstract

The aim of this study was to develop an in vitro human peripheral lymphocyte micronucleus bioassay involving phytohemagglutinin stimulant for urines containing adriamycin (ADR) and cyclophosphamide (CP). In vitro studies with defined concentrations of ADR, CP, and fresh urine showed that mitotic indices and micronuclei counts/1,000 cells had to be log (X + 1) transformed to be able to use parametric statistics and that a specific micronucleus assay for ADR in the presence of CP and urine for 5-15 ng ADR/mL had been developed. Whereas CP alone could be detected between 196-522 micrograms/mL, this effect was abolished in the presence of 15 ng ADR/mL. Interdonor variabilities relative to ADR sensitivity and CP linear dynamic range were marked, but intradonor variability was small. The MN bioassay tolerated < 10% urine. Results for urines from nine patients receiving antineoplastic drugs (CP, all; ADR, 3; 5-fluorouracil, 3; methotrexate, 3; vincristine, 4; procarbazine, 1; and megestrol acetate, 1) showed that only 1/3 patients given ADR were detected, and two others not given ADR were positive. All frozen urines from the 12 control subjects and the nine patients exhibited depressed mitotic index, with, however, no control patient urines inducing increased micronuclei. Two patients had urines of undefined genotoxic potential since undepressed mitotic indices were not attainable by dilution. The effects of combination chemotherapy in addition to freezing and storage influences were complex. More research is required to be able to interpret the results.

Bael, Aegle marmelos (L.) Correa, an Indian medicinal plant protects V79 cells against the genotoxic effect of doxorubicin

“…An identical effect has been observed in the present study, where DOX induced a concentration dependent rise in the MNBNCs in V79 cells exposed to various concentrations of DOX and the dose response was linear quadratic. A dose dependent increase in the frequency of micronuclei has been reported in vitro and in vivo (Boucher et al, 1993;Jagetia and Nayak, 1996;Jagetia and Aruna, 2000;. Our findings that DOX increased the frequency of binucleate cells bearing two and multiple MN are in agreement with the earlier reports, where a similar effect has been observed Takahashi, 1998, 1999;Al Harbi, 1993;Jagetia andNayak, 1996, 2000;Jagetia and Aruna, 2000;Venkatesh et al, 2007).…”

Modulation of Doxorubicin-Induced Genotoxicity by Aegle marmelos in Mouse Bone Marrow: A Micronucleus Study

“…DOX treatment has been shown to increase the frequency of MN in mice bone marrow and human lymphocytes. 12,54 Reports regarding the reduction in DOXinduced MN by AME are unavailable, and this is probably the first report in which AME pretreatment has been found to reduce DOX-induced MN in vivo. However, earlier studies have shown that AME pretreatment reduces the frequency of radiation-induced MN in cultured human peripheral blood lymphocytes as well as radiation-induced MN in mice bone marrow cells, 55,56 indicating that it does not exert a genotoxic effect in the dose range studied.…”

“…The acentric fragments give rise to MN with an efficiency of about 80%, 54 and this apparent saturation of the micronucleus yield is due to the fact that after cell division, the acentric fragment is likely to produce a micronuclei in only 1 of the 2 daughter cells and not because of low efficiency of MN formation. This fact alone will reduce the frequency of MN by a factor of for each cell division, and there is also the possibility of incorporation of more than 1 fragment during the formation of MN.…”