Chung Young-Shin, Pak Bumsoo, Han Sehee, Lee Jiyeon, Kim Jiyoung, Back Seng-Min, Park Cho-Rong, Kim Su-Hwan, Lee Jong-Kwon
Department of Biotechnology and The Research Institute for Basic Sciences, Hoseo University, Asan, Chungnam, 31499, Republic of Korea.
Department of Biotechnology and The Research Institute for Basic Sciences, Hoseo University, Asan, Chungnam, 31499, Republic of Korea.
Mutat Res Genet Toxicol Environ Mutagen. 2018 Jul;831:62-68. doi: 10.1016/j.mrgentox.2018.05.015. Epub 2018 May 26.
The mutagenic potencies of 1,3-propane sultone (PS), N-propyl-N-nitrosourea (PNU), and mitomycin C (MMC) were investigated in three independent laboratories in Korea using the Pig-a assay in vivo. Sprague-Dawley rats were treated with vehicle or test substance on three consecutive days. Blood samples were collected for measuring Pig-a mutant phenotypes (CD59-deficient erythrocytes, RBC; CD59-deficient reticulocytes, RET) on days -1, 15, and 29 after the first treatment. In some studies, blood was collected for determining DNA damage (comet assay) on day 3 and measuring micronucleated reticulocytes (MN-RET) on day 4. Treatment with the alkylating agents PS and PNU induced dose-dependent increases in the frequency of RBC on days 15 and 29, and caused maximum elevations in the frequency of RET on day 15. Inter-laboratory comparison of the day 29 Pig-a assay data confirmed the mutagenic potencies of PS and PNU, and showed good agreement among the test sites. Treatment with the DNA cross-linker MMC induced increases in the frequencies of RBC and RET on days 15 and 29 (all three laboratories). MN-RETs increased significantly in animals treated with PS, PNU, or MMC, but biologically significant increases in DNA damage were observed only with PS and PNU, and not with MMC. The results of this study indicate that the Pig-a assay is a sensitive, reproducible method for evaluating the in vivo mutagenicity of various test substances, in particular, DNA cross-linkers and alkylating agents. Our limited data on integrating the Pig-a assay with the comet and micronucleus assays indicate that a short-term treatment protocol evaluating these three endpoints in a single set of animals may be a robust strategy for evaluating in vivo genotoxicity.
在韩国的三个独立实验室中，使用体内Pig-a试验研究了1,3-丙烷磺酸内酯（PS）、N-丙基-N-亚硝基脲（PNU）和丝裂霉素C（MMC）的诱变潜力。将Sprague-Dawley大鼠连续三天用赋形剂或受试物质处理。在首次处理后的第-1天、第15天和第29天采集血样，以测量Pig-a突变表型（CD59缺陷红细胞，RBC；CD59缺陷网织红细胞，RET）。在一些研究中，在第3天采集血液以测定DNA损伤（彗星试验），并在第4天测量微核网织红细胞（MN-RET）。用烷化剂PS和PNU处理可导致第15天和第29天RBC频率呈剂量依赖性增加，并在第15天导致RET频率最大升高。第29天Pig-a试验数据的实验室间比较证实了PS和PNU的诱变潜力，并显示各试验点之间具有良好的一致性。用DNA交联剂MMC处理可导致第15天和第29天RBC和RET频率增加（所有三个实验室）。用PS、PNU或MMC处理的动物中MN-RET显著增加，但仅在PS和PNU处理组中观察到具有生物学意义的DNA损伤增加，而MMC处理组未观察到。本研究结果表明，Pig-a试验是一种灵敏、可重复的方法，用于评估各种受试物质的体内诱变性，特别是DNA交联剂和烷化剂。我们关于将Pig-a试验与彗星试验和微核试验相结合的有限数据表明，在一组动物中评估这三个终点的短期处理方案可能是评估体内遗传毒性的有力策略。
