ENU致大鼠体内Pig-a基因突变的时-效关系和量-效关系研究

doi:10.3969/j.issn.1004-616x.2013.03.008

Abstract

Abstract:

OBJECTIVE: To study time-course and dose-response relationship of ENU-induced rat phosphatidylinositol glycan class-A(Pig-a) gene mutation and explore the potential of this assay to be integrated into repeat dose study，and to establish a rat Pig-a gene mutation assay. MEHTODS：Fifteen rats were randomly assigned to be treated with PBS and ENU (20 mg/kg and 40 mg/kg) for 3 consecutive days by oral gavage. Jugular blood samples were collected on days -1 (the day before administration)，15，30 and 45. Erythrocytes were enriched and incubated with Anti-CD59-PE and nucleic acid dye solution，and then analyzed with flow cytometer. RESULTS：On days 15，30 and 45，the Pig-a gene mutation frequencies in RBCs and RETs in low and high dosage groups were elevated significantly compared with that in control group (all P values <0.01)，and the mean frequencies in high dosage group were about 2-3 times of that in low dosage group. The Pig-a gene mutation frequencies were maintained at high levels during days 15-45. CONCLUSION：The in vivo Pig-a gene mutation assay was established successfully in rats. Time-course results suggested that this assay can be integrated into repeated-dose study.

Key words: Pig-a gene, mutation, time-course relationship, dose-response relationship

ZHANG Ming，ZHOU Chang-hui，WANG Zheng，WANG Qing-li，CHANG Yan，. Time-course and dose-response relationship study of ENU-induced Pig-a gene mutation in the rat[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2013.03.008.

References

[1] ICH. Guidance on genotoxicity testing and data interpretation for pharmaceuticals intended for human use [EB/OL]. [2011-11-09]. http：//www.ich.org/products/guidelines/safety/article/ safety- guidelines.html.

[2] Lambert IB，Singer TM，Boucher SE，et al. Detailed review of transgenic rodent mutation assays [J]. Mutat Res，2005，590 (1/2/3)：1-280.

[3] Dearfield KL，Veronique T，Cimino MC，et al. Follow-up actions from positive results of in vitro genetic toxicity testing [J]. Environ Mol Mutagen，2011，52 (3)：177-204.

[4] Takeda J，Miyata T，Kawagoe K，et al. Deficiency of the GPI anchor caused by somatic mutation of Pig-a gene in paroxysmal nocturnal hemoglobinuria[J]. Cell，1993，73 (4)：703–711.

[5] Kawagoe K，Takeda J，Endo Y，et al. Molecular cloning of murine Pig-a，a gene for GPI-anchor biosynthesis，and demonstration of interspecies conservation of its structure， function，and genetic locus[J]. Genomics，1994，23 (2)： 566–574.

[6] Dobrovolsky VN，Miura D，Heflich RH，et al. The in vivo Pig-a gene mutation assay，a potential tool for regulatory safety assessment[J]. Environ Mol Mutagen，2010，51 (8/9)：825-835.

[7] Dertinger SD，Phonethepswath S，Franklin D，et al. Integration of mutation and chromosomal damage endpoints into 28-day repeat dose toxicology studies[J]. Toxicol Sci，2010，115 (2)： 401–411.

[8] Dertinger SD，Phonethepswath S，Weller P，et al. International Pig-a gene mutation assay trial：evaluation of transferability across 14 laboratories[J]. Environ Mol Mutagen，2011，52 (9)： 690-698.

[9] Bhall JA，Shaddock JG，Pearce MG，et al. Report on stage III Pig-a mutation assays using benzo[a]pyrene[J]. Environ Mol Mutagen， 2011，52 (9)：731-737.

[10] Miura D，Dobrovolsky VN，Kimotoa T，et al. Accumulation and persistence of Pig-a mutant peripheral red blood cells following treatment of rats with single and split doses of N-ethyl-N-nitrosourea[J]. Mut Res，2009，677 (1/2)：86–92.

[1]	XU Qilu, GAO Hui, RAN Xuehong, WANG Junjun, ZHANG Junying, LIU Liping. Gene mutation and clinical correlation analyses of myeloid leukemia with abnormal karyotypes [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(4): 296-301,306.
[2]	SHI Hailin, HE Tianji, LIU Feng, CAI Menghui, GE Bo. Prognostic value of TCGA-database mutation burden in muscle-invasive bladder cancers [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(6): 430-437,443.
[3]	ZHANG Jiahong, ZHANG Qingying, ZHANG Yanhong, CHEN Jialian, XIE Fang, LIU Ruiguo, WU Rong, WU Longfei. Screening and evaluation of gene mutations in a familial hypercholesterolemia family [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(3): 229-232,245.
[4]	CAO Yiyi, LIU Weiying, YOU Xinyue, XI Jing, Chen Ruixue, ZHANG Xinyu, LUAN Yang. In vitro genotoxicity evaluation of BDE-3, BDE-47 and BDE-209 [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(1): 15-20,28.
[5]	ZHENG Kai, WANG Bingyu, XU Xinyun, GENG Hong, HUANG Haiyan, LIU Wei, LONG Dingxin. Mutagenicity of PM_2.5 samples from Shenzhen and Taiyuan [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(6): 483-487.
[6]	YANG Jinru, QIN Lulu, ZHU Yongfei. Meta analysis on association between JAK2 gene mutation and susceptibility to chronic myeloid leukemia [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(3): 231-237.
[7]	LI Ruowan, ZHOU Changhui, HUANG Pengcheng, CHANG Yan. Establishment of an in vitro PIG-A gene mutation assay based on TK6 cells for genotoxicity tests [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(3): 242-248.
[8]	ZHANG Feng, ZHAO Long, FAN Jinping, WU Xueling, MENG Shufang. Induction of tumor in NIH mice by the plasmid containing the V112A and Q61R mutated HRAS [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 1-8.
[9]	SHANG Pingping, HUA Chenfeng, XIE Fuwei, LI Xiang. Mutagenicity of cigarette samples with different tar releases in the TK gene mutation assay [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 49-52.
[10]	WANG Ping, LIU Bingfeng, YANG Yunfeng, CAI Yuwei, TAN Na, YAO Lijuan, LUO Ranran, ZHANG Guo. Mutation frequencies of tumor-related genes in the pancreatic tissues of diet-induced obese mice [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(3): 182-187.
[11]	XIA Ying, FU Shaohua, ZHANG Yinjing, WANG Wei. Genetic toxicity test of a pesticide, Paichongding [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(2): 136-139.
[12]	GUO Xiao, WANG Rui, WU Juan, JI Xiaokun, WANG Heng, ZHANG Yan, MA Yang, DU Yun. Significance of EGFR gene mutation in fresh cytological specimens of lung adenocarcinoma [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(1): 67-70.
[13]	CHANG Chen, ZHAO Xiaohan, JIANG Ye, XU Xin, CAI Yan, PEI Yuhui, ZHANG Yu, HAO Jiajie, WANG Mingrong. Identification of predominant genome DNA alterations and their clinical implication in esophageal squamous cell carcinoma [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(1): 1-6.
[14]	MENG Xiaoqing, FANG Zhijia, WEI Hongyan, YANG Zhanju, HUANG Zhiwei. Genotoxicity investigation of tri- and hexavalent chromium using a yeast SUP4-o mutation assay [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(1): 65-69.
[15]	YUAN Ye, CHEN Dandan, JING Shufang, YU Yongsheng, CHEN Jianheng, WU Chunqi, SHI Chang. Mutagenic effects of N-(n-butyl)-thiophosphorictriamide in somatic and germ cells of mice [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(6): 432-437.

Time-course and dose-response relationship study of ENU-induced Pig-a gene mutation in the rat

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments