食品中甲醛的细胞毒性及基因表达影响  

Research of Cytotoxicity and Gene Expression by Formaldehyde in the Food

作　　者：刘华 

机构地区：[1]广州工商学院,广东广州510000

出　　处：《食品与生物技术学报》2017年第6期625-634,共10页Journal of Food Science and Biotechnology

基　　金：广东省教育科研"十二五"规划2013年度研究项目(2013JK313)

摘　　要：为了探讨食品中甲醛的细胞毒作用,检测不同浓度下甲醛对细胞生长的抑制情况,并运用基因芯片技术检测甲醛对淋巴细胞基因表达的影响,筛选并分析与甲醛毒性和致癌性相关的差异表达基因。作者进行MTT实验,绘制淋巴细胞的细胞生长曲线及细胞形态观察,然后用5×10-5mol/L甲醛处理人外周血淋巴细胞24 h,提取实验组和对照组细胞m RNA,经逆转录和标记后与人类全基因组芯片(约35 000个点)杂交,芯片扫描后提取并处理数据,筛选出甲醛诱导显著改变的基因,并进行生物信息学分析。发现甲醛与淋巴细胞增殖存在的剂量-效应关系,验证了甲醛对淋巴细胞的细胞毒作用;共筛选出显著上调基因211个,显著下调基因113个,分别参与氧化应激、细胞凋亡、核酸代谢、细胞周期调控、肿瘤生成、肿瘤抑制等途径,推断出OXR1、MLH1、TP53等基因表达的改变可作为甲醛毒性和致癌性的遗传学基础。本研究获得的数据为进一步阐明甲醛的毒性和致癌机理,检测食品中甲醛暴露的早期生物学标志提供了理论依据,证明了基因芯片技术在食品毒理学研究中的可行性和优越性,为建立新的有毒物质暴露的检测和研究方法提供了实验依据。To investigate the cytotoxic effects of formaldehyde in the food,we detect different concentrations of formaldehyde on the inhibition of cell growth, and use microarray technology to detect the influence of lymphocyte gene expression by formaldehyde in order to screen and analyze the differentially expressed genes related to toxicity and carcinogenicity of formaldehyde. By making MTT experiments, the cytotoxicity of formaldehyde was verified by comparing lymphocytes＇ growth curve and morphology. Peripheral blood lymphocytes were treated with 5 × 10^5 mol/L formaldehyde for 24 h.The total RNA from cells of the subjects of the treated group and the control group were isolated,then they were reversely transcribed and labeled with fluorescence dyes,and finally hybridized with human whole genome microarray （about 35 000 spots）. Microarray data wasextracted after microarray scanning,and differentially expressed genes induced by formaldehyde were screened and analyzed by bioinformatics. In MTT experiments,human peripheral blood lymphocytes were treated with various doses of formaldehyde. The dose-effect relationship was also observed after treatment with high doses. 211 up-regulated and 113 down-regulated genes were screened after scanning and data extracting. Genes which significantly altered were represented by several categories,including oxidative stress,DNA repair,apoptosis,nucleic acid metabolism,cell cycle, oncogenesis, tumor suppression and so on. The result suggested the differential expression of these genes of OXR1 ,MLH1,TP53 may be the genetic basis of formaldehyde toxicity and carcinogenicity. The study provided theoretical evidence for elucidating the toxic and carcinogenic mechanism, detecting of the biomarker for formaldehyde exposure, and preventing the formaldehyde poisoning. It also verified the microarray technique is the feasible and advantageous in food toxicological research. It provides an experimental basis for the establishment of new methods to detect and study exposure of toxic substan

关 键 词：甲醛 细胞毒性 细胞生长曲线 基因表达 

分 类 号：TS201[轻工技术与工程—食品科学]