Abstract：Objective To explore the mutagenic effect of sodium pentachlorophenate (NaPCP) on zebrafish p53 gene coding sequence(CDS) in somatic cell.Methods The experiment was carried out using tuebingen strain of zebrafish, according to the results of acute toxicity test to determine the exposure levels in zebrafish. Zebrafish were randomly divided into blank control group and exposed groups, each containing 10 zebrafish. After exposing for 45d of NaPCP, the RNA was extracted from liver of zebra fish, and the p53 gene including a complete coding sequence of was obtained by RT-PCR.Results LC50 of NaPCP was 18.4 μg/L.Sequence analysis showed that the p53 gene CDS length of 1125bp, encoding 374 amino acids. The percent identity between the published zebrafish sequence of p53 (GI:425876786)and ours was 99.2％,with the other biological sequence of p53 existing some differences. After 45d exposure, zebrafish p53 gene of NaPCP exposure group had mutated at the concentration of 1.8 μg /L. The base substitution of GAG→AAG at codon 8,CAT→CAG at codon 148 and CAG→CAA at codon 229 were detected by PCR-directed sequencing. This may result in the Glu→Lys and His→Gln of expressed p53 proteinConclusion NaPCP is a kind of gene mutation, which can induce the mutation of p53 gene in zebrafish somatic cells, that has the potential mutagenic risk for humans.
夏勇, 徐彩菊, 傅剑云, 宋燕华, 蔡德雷, 孟真. 五氯酚钠诱导斑马鱼体细胞p53基因突变的克隆测序研究[J]. 预防医学, 2017, 29(4): 325-329,333.
XIA Yong, XU Cai-ju, FU Jian-yun, SONG Yan-hua, CAI De-lei, MENG Zhen. A study on the cloning and sequencing of mutation of p53 gene induced by sodium pentachlorophenate in somatic cell of zebrafish. Preventive Medicine, 2017, 29(4): 325-329,333.
[1]尹晓晨,曾明,王春香, 等.五氯酚钠对雄性小鼠的生殖毒性作用[J].生态毒理学报,2008,3(5):473-477. [2]RYLANDER C, LUND E, FRYLAND L, et al. Predictors of PCP, OH-PCBs, PCBs and chlorinated pesticides in a general female Norwegian population[J].Environ Int,2012,43(1):13-20. [3]杨淑贞,韩晓冬,陈伟.五氯酚对生物体的毒性研究进展[J].环境与健康杂志,2005,22(5):396-398. [4]CHENG P,ZHANG Q, SHAN X, et al.Cancer risks and long-term community-level exposure to pentachlorophenol in contaminated areas, China[J].Environ Sci Pollut Res Int,2015,22(2):1309-1317. [5]陈坤,赵玉婉,马新源,等.浙江省嘉善县人群结直肠癌发病与有机氯污染的关系[J]. 中华流行病学杂志, 2004,25(6):479-483. [6]VOGELSTEIN B, LANE D, LEVINE A. Surfing the p53 network[J]. Nature,2000,408(6810):307-310. [7]HU S , ZHAO L , YANG J , et al. The association between polymorphism of p53 Codon72 Arg/Pro and hepatocellular carcinoma susceptibility: evidence from a meta-analysis of 15 studies with 3,704 cases [J]. Tumour Biol, 2014, 35 (4): 3647-3656. [8]LANGHEINRICH U,HENNEN E,STOTT G,et al. Zebrafish as a model organism for the identification and characterization of drμgs and genes affecting p53 signaling[J]. Curr Biol, 2002,12(23): 2023-2028. [9]AMANUMA K,TAKEDA H,AMANUMA H,et al.Transgenic Zebrafish for Detecting Mutations Caused by Compounds in Aquatic Environments[J]. Nat Biotechnol,2000,18(1): 62-65. [10]夏勇,郑云燕,宋燕华,等.百草枯对斑马鱼的生理指标和胚胎发育影响[J].环境与职业医学,2011,28(5):271-276. [11]BUSQUET F , STRECKER R , RAWLINGS J M, et al. OECD validation study to assess intra- and inter-laboratory reproducibility of the zebrafish embryo toxicitytest for acute aquatic toxicitytesting [J]. Regul Toxicol Pharmacol ,2014, 69 (3): 496-511. [12]国家环境保护总局《水和废水监测分析方法》编委会. 水和废水监测分析方法[M]. 4版.北京:中国环境科学出版社,2006:780. [13]王坤元,刘莉,戴文聪,等. 二乙基亚硝胺诱导建立斑马鱼肝纤维化模型[J]. 南方医科大学学报,2014,34(6):777-782. [14]沈骅,王晓蓉,张景飞,等. Cu2+和Cu-EDTA对鲫鱼脑组织应激蛋白HSP70诱导的影响[J]. 环境科学,2004,25(3): 94-97. [15]张尤恩.有毒化学物质毒性手册[M].北京:北京医科大学、中国协和医科大学联合出版社,1993:162. [16]中华人民共和国卫生和计划生育委员会. 哺乳动物红细胞微核试验:GB15193.5—2014[S]. 2014:1-4. [17]宋瑞霞,阮鸿洁,刘征涛,等. 五氯酚钠致突变性研究[J]. 中国公共卫生,2008,24(3): 337-339. [18]YIN D, GU Y, LI Y, et al. Pentachlorophenol treatment in vivo, elevates point mutation rate in zebrafish p53 gene[J]. Mutation Research/fundamental & Molecular Mechanisms of Mutagenesis, 2006, 609(1):92-101. [19]许文青,樊柏林,陈明,等.五氯苯酚和五氯苯酚钠毒性作用研究进展[J].中国药理学与毒理学杂志,2011,25(6):596-599. [20]王心如. 毒理学基础[M].6版.北京:人民卫生出版社,2012,153-154. [21]石年.卫生毒理学[M]. 北京:人民卫生出版社,2002,273-274.
