Procarbazine genotoxicity in the Muta™Mouse; strong clastogenicity and organ-specific induction of lacZ mutations

Suzuki, Takayoshi; Uno, Yoshifumi; Idehara, Kenji; Baba, Tsuneo; Maniwa, Jiro; Ohkouchi, Akiko; Wang, Xue; Hayashi, Makoto; Sofuni, Toshio; Tsuruoka, Masaki; Miyajima, Hirofumi; Kono, Koji

doi:10.1016/s1383-5718(99)00060-1

Cited by 22 publications

(21 citation statements)

References 36 publications

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“…In this study, PCH's in vivo genotoxic activity was readily apparent, as each of the endpoints investigated showed statistically significant effects at all of the doses tested. As also found by Suzuki et al (), we observed potent clastogenic activity as evidenced by robust MN induction. Whereas Suzuki and colleagues failed to detect significant increases in in vivo lacZ mutations in bone marrow, liver, testis, spleen, kidney, and lung upon single administration of 50 mg PCH/kg, they did observe significant lacZ mutation induction at 150 mg/kg/day when administered for five consecutive days (cumulative dose: 750 mg/kg) particularly in those tissues that are prone to develop tumors following PCH exposure (i.e., lung, bone marrow, and spleen).…”

Section: Discussionsupporting

confidence: 90%

“…In contrast, PCH mutagenicity was evident in lung, bone marrow, and spleen after five daily administrations of 150 mg/kg by i.p. injection (Suzuki et al ). Following a 3‐day exposure regimen by oral gavage Phonethepswath et al () confirmed the strong clastogenic activity of PCH and observed a moderate but variable mutagenic response using the rodent phosphatidylinositol glycan‐class A ( Pig‐a ) assay.…”

Section: Introductionmentioning

confidence: 99%

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Integrated In Vivo Genotoxicity Assessment of Procarbazine Hydrochloride Demonstrates Induction of Pig‐a and LacZ Mutations, and Micronuclei, in MutaMouse Hematopoietic Cells

Maurice

Dertinger

Yauk

et al. 2019

Environ and Mol Mutagen

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Procarbazine hydrochloride (PCH) is a DNA‐reactive hematopoietic carcinogen with potent and well‐characterized clastogenic activity. However, there is a paucity of in vivo mutagenesis data for PCH, and in vitro assays often fail to detect the genotoxic effects of PCH due to the complexity of its metabolic activation. We comprehensively evaluated the in vivo genotoxicity of PCH on hematopoietic cells of male MutaMouse transgenic rodents using a study design that facilitated assessments of micronuclei and Pig‐a mutation in circulating erythrocytes, and lacZ mutant frequencies in bone marrow. Mice were orally exposed to PCH (0, 6.25, 12.5, and 25 mg/kg/day) for 28 consecutive days. Blood samples collected 2 days after cessation of treatment exhibited significant dose‐related induction of micronuclei in both immature and mature erythrocytes. Bone marrow and blood collected 3 and 70 days after cessation of treatment also showed significantly elevated mutant frequencies in both the lacZ and Pig‐a assays even at the lowest dose tested. PCH‐induced lacZ and Pig‐a (immature and mature erythrocytes) mutant frequencies were highly correlated, with R 2 values ≥0.956, with the exception of lacZ vs. Pig ‐ a mutants in mature erythrocytes at the 70‐day time point (R 2 = 0.902). These results show that PCH is genotoxic in vivo and demonstrate that the complex metabolism and resulting genotoxicity of PCH is best evaluated in intact animal models. Our results further support the concept that multiple biomarkers of genotoxicity, especially hematopoietic cell genotoxicity, can be readily combined into one study provided that adequate attention is given to manifestation times. Environ. Mol. Mutagen. 60:505–512, 2019. © 2018 Her Majesty the Queen in Right of Canada

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Integrated In Vivo Genotoxicity Assessment of Procarbazine Hydrochloride Demonstrates Induction of Pig‐a and LacZ Mutations, and Micronuclei, in MutaMouse Hematopoietic Cells

Maurice

Dertinger

Yauk

et al. 2019

Environ and Mol Mutagen

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“…Procarb's in vivo genotoxic activity was readily apparent in this study that used flow cytometry‐based methods to monitor MN‐RET and Pig‐a mutant cell frequencies. As also found by Suzuki et al [], we observed potent clastogenic activity as evidenced by robust induction of micronuclei. Whereas Suzuki et al failed to detect significant in vivo lacZ mutation in bone marrow, liver, testis, spleen, kidney, and lung upon single administration of 50 mg Procarb/kg, they did observe significant lacZ induction at 150 mg/kg/day when administered for 5 consecutive days.…”

Section: Discussionsupporting

confidence: 89%

Flow cytometric analysis of Pig‐a gene mutation and chromosomal damage induced by procarbazine hydrochloride in CD‐1 mice

Phonethepswath

Avlasevich

Torous

et al. 2013

Environ and Mol Mutagen

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Procarbazine is a genotoxic carcinogen whose DNA-damaging activities are not reliably detected in vitro. We evaluated the in vivo genotoxic effects of procarbazine on hematopoietic cells of male CD-1 mice using a multi-endpoint study design that scored micronucleated reticulocyte (MN-RET) frequency and gene mutation at the Pig-a locus. CD-1 mice were treated for 3 days with procarbazine, up to 150 mg/kg/day. Blood samples collected on Day 3 exhibited robust induction of MN-RETs, with the high dose group exhibiting a mean 29-fold increase. Blood collected 15 and 30 days after treatment began was analyzed for Pig-a mutation with a dual labeling method that facilitated mutant cell frequency measurements in both total erythrocytes and the reticulocyte subpopulation. Procarbazine significantly increased mutant reticulocyte frequencies by Day 15. Mutant erythrocyte responses were also apparent, with a peak incidence observed for the high dose group on Day 30. These results demonstrate that the complex metabolism and resulting genotoxicity of procarbazine is best evaluated in intact animal models, and show that the flow cytometric methods employed offer a means to efficiently monitor both in vivo chromosomal damage and mutation.

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“…Premeiotic exposure to 4 mg/kg of mitomycin C did not affect the frequency of gpi-y mutation in the germ line of transgenic male mice (40); in contrast, our data show that this anticancer drug is mutagenic at doses of V2.5 mg/kg. It was previously reported that doses of procarbazine as high as 750 mg/kg did not affect mutation rates at the lacZ gene in testicular DNA of MutaMouse transgenic males (49), yet in our study, a highly significant increase in ESTR mutation frequency was detected in the germ line of male mice exposed to just 40 mg/kg of procarbazine. Taken together, these data clearly show that the sensitivity of SM-PCR techniques to a wide range of chemical mutagens and anticancer drugs substantially exceeds that of a variety of mouse transgenic assays.…”

Section: Discussioncontrasting

confidence: 72%

Single-Molecule PCR Analysis of Germ Line Mutation Induction by Anticancer Drugs in Mice

2008

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Understanding and estimating the genetic hazards of exposure to chemical mutagens and anticancer drugs in humans requires the development of efficient systems for monitoring germ line mutation. The suitability of a single-molecule PCRbased approach for monitoring mutation induction at the mouse expanded simple tandem repeat (ESTR) locus Ms6-hm by chemical mutagens and anticancer drugs has been validated. The frequency of ESTR mutation was evaluated in the germ line of male mice exposed to the well-characterized alkylating agent and mutagen, ethylnitrosourea, and four widely used anticancer drugs, bleomycin, cyclophosphamide, mitomycin C, and procarbazine. The dose-response of ethylnitrosourea-induced mutation was found to be very close to that previously established using a pedigree-based approach for ESTR mutation detection. Paternal exposure to the clinically relevant doses of bleomycin (15-30 mg/kg), cyclophosphamide (40-80 mg/kg), and mitomycin C (2.5-5 mg/kg) led to statistically significant, dose-dependent increases in ESTR mutation frequencies in the germ line of treated male mice. Exposure to procarbazine led to a maximal increase in mutation frequency at 50 mg/kg, with a plateau at the higher concentrations. The results of this study show that the singlemolecule PCR technique provides a new and efficient experimental system for monitoring the genetic effects of anticancer drugs, capable of detecting increases in mutation rates at clinically relevant doses of exposure. In addition, this approach dramatically reduces the number of mice needed for the measurement of germ line mutation induction.

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Procarbazine genotoxicity in the Muta™Mouse; strong clastogenicity and organ-specific induction of lacZ mutations

Cited by 22 publications

References 36 publications

Integrated In Vivo Genotoxicity Assessment of Procarbazine Hydrochloride Demonstrates Induction of Pig‐a and LacZ Mutations, and Micronuclei, in MutaMouse Hematopoietic Cells

Integrated In Vivo Genotoxicity Assessment of Procarbazine Hydrochloride Demonstrates Induction of Pig‐a and LacZ Mutations, and Micronuclei, in MutaMouse Hematopoietic Cells

Flow cytometric analysis of Pig‐a gene mutation and chromosomal damage induced by procarbazine hydrochloride in CD‐1 mice

Single-Molecule PCR Analysis of Germ Line Mutation Induction by Anticancer Drugs in Mice

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