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Cytotoxicity of acrylamide and its epoxide glycidamide in CHO cells expressing human cytochrome P450 2E1

Journal of Nanjing Medical University 2006年第6期20卷 325-330页

作者： Shoulin Wang Xiaoyang He Xinru Wang Junyan Hong Institute of Toxicology Nanjing Medical University Nanjing 210029 China School of Public Health University of Medicine and Dentistry of New Jersey Piscataway NJ 08854 USA

Objective： To investigate whether CYP2E1 is responsible for the acrylamide metabolic activation in FIp-In CHO cell system. Methods： CYP2E1 cDNA was subcloned from the human liver full-length cDNA library and subsequently transfected into the FIp-In CHO cells to generate the stable transfectant of CYP2E1. The CYP2E1 mRNA expression was determined by RT-PCR. Acrylamide and its epoxide glycidamide induced cytotoxicity and cell cycle arrest in G2/M were conducted using MTS assay and flow cytometry, respectively. Results： In the CHO cell stably expressing CYP2E1 （CHO-2E1）, a -1.5 kb size of band was detected from the mRNA in the cells while no corresponding band in the CHO-vector cells, which indicated that CYP2E1 was successfully transfected in the CHO cells. Compared with the CHO-vector cells, acrylamide showed a concentration dependent loss of viability in the CHO-2E1 cells but no significant change of G2/M arrest was found. As expected, glycidamide induced similar profile of cytotoxicity in both of the cells, and G2/M arrest presented a concentration-dependent increased in the CHO-2E1 cells. Conclusion： The result suggested that CYP2E1 might be responsible for the acrylamide metabolism, and its metabolite glycidamide was a direct cytotoxic and genotoxic agent. It should be further considered whether acrylamide-induced toxicity is through its epoxide glycidamide in the presence of CYP2E1.

关键词： Flp-In CliO cells cytochrome P450 2El cytotoxicity acrylamide glycidamide

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Effects of glycidamide on Activity and Progesterone Biosynthesis of Rat R2C Leydig Cells

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Agricultural Science & Technology 2015年第6期16卷 1187-1190页

作者：许伟孙建霞白顺李名薇文罗娜刘洁生李宏业黄亚东白卫滨暨南大学生命科学学院食品科学与工程系广东广州510632 广东工业大学轻工化工学院广东广州510006

Objective] This study aimed to investigate the effects of glycidamide （GA） on growth and progesterone biosynthesis of rat R2C Leydig cel s cultured in vitro. [Method] The R2C Leydig cel s were treated with GA with concentrations of 0.25, 0.5, 0.75, 1, 2, 4 and 6 mmol/L respectively for 48 h. The IC25, IC50 and IC75 values of GA were al detected by MTT assay. The Leydig cel s were treated with GA at concentrations of IC25, IC50 and IC75 respectively for 48 h, and then the morphology of Leydig cel s was observed. After the Leydig cel s were treated with GA for 4 h, the cel ular DNA damage was measured by the comet assay technique; and after the Leydig cel s were with treated with GA for 24 h, the progesterone biosynthesis amount was detected by radioimmunoassay （RIA）. [Result] GA could inhibit the via-bility of R2C Leydig cel s, and its IC25, IC50 and IC75 were 0.635, 0.872 and 1.198 mmol/L, respectively. The GA at concentrations of IC25, IC50 and IC75 affected the growth and morphology of rat R2C Leydig cel s in varying degrees. The 4-h treat-ment of GA could significantly damage the DNA of R2C Leydig cel s, and the 24-h treatment of GA at concentrations of IC25, IC50 and IC75 al reduced the progesterone biosynthesis amount. [Conclusion] GA could inhibit the growth and progesterone biosynthesis of rat R2C Leydig cel s.

关键词： glycidamide R2C Leydig cel s Cel activity DNA damage Proges-terone

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Study of N7-（2-carbamoyl-2-hydroxyethyl）Guanine in Urine Samples Repeatedly Collected from Acrylamide-Exposed Workers

Study of N7-（2-carbamoyl-2-hydroxyethyl）Guanine in Urine S...

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第八届海峡两岸毒理学研讨会

作者： Yu-Fang Huang Mei-Lien Chen Chih-Chun Huang Saou-Hsing Liou Su-Yin Chiang Kuen-Yuh Wu Institute of Environmental and Occupational Health Sciences National Yang Ming University Department of Food Science National Pingtung University of Science and Technology Division of Environmental Health and Occupational Medicine National Health Research Institutes School of Chinese Medicine China Medical University Institute of Occupational Medicine and Industrial Hygiene National Taiwan University

Acrylamide（AA）is a widely-used industrial chemical,present in tobacco smoke and high-temperature processed foods,and classified as a probable human *** activation from AA to glycidamide（GA）to form DNA adducts is critical to AA carcinogenicity.N7-（2-carbamoyl-2-hydroxyethyl）guanine（N7-GAG）,the most abundant DNA adduct induced by GA,was recently detected in urine of smokers and *** study aims to analyze N7-GAG in urine samples repeatedly collected from AA-exposed workers to explore its elimination ***,8 exposed and 36 control workers were recruited for this ***-and post-shift urine samples were collected from the exposed group in parallel with personal sampling for 8 consecutive days and from the control group for one day.N7-GAG was analyzed with our newly-developed two-step SPEs and on-line SPE liquid chromatography-electronspray ionization/tandem mass spectrometry（LC-ESI-MS/MS）***-and GA-mercapturic acids（AAMA and GAMA）were also analyzed to serve as biomarkers for AA current *** exposed workers excreted significantly higher N7-GAG levels than the controls.N7-GAG was significantly associated with AAMA and GAMA,*** of the N7-GAG data with linear mixed-effect model reveals a mean half-life at 346.5 hrs with a 95%confidence interval（140.3-737.2）*** from this study suggest that urinary N7-GAG levels could be elevated by occupational exposures to AA and intra-and inter-individual differences were damped with the half-life of the biomarkers.

关键词： Acrylamide N7-(2-carbamoyl-2-hydroxyethyl)guanine glycidamide elimination kinetics

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