杀草强对人甲状腺细胞系Nthy-ori-3-1细胞全基因表达谱的影响
发布时间：2018-10-16 14:27
【摘要】：目的甲状腺是人体最大的内分泌腺,其分泌的甲状腺激素对人体的生长发育和新陈代谢有很重要的作用。甲状腺干扰物对人及动物的甲状腺有很显著的影响,能够干扰甲状腺功能、诱发自身免疫性甲状腺疾病、还有潜在的致甲状腺肿瘤的作用。在已知的甲状腺干扰物中,农药占有很大的比例。杀草强作为一种非选择性除草剂,被证实能够干扰甲状腺激素的合成以及导致甲状腺增生和甲状腺肿瘤,但是对其作用机制的研究报道很少。本研究以人甲状腺细胞系Nthy-ori-3-1细胞为受试对象,在排除细胞毒性作用的剂量下,从基因水平探索杀草强致甲状腺肿瘤相关的作用机制。方法实验设计了杀草强0μg/ml、μg/ml.10pg/ml.100μg/ml四个剂量组,用含相应受试物的培养基培养人甲状腺细胞系Nthy-ori-3-1细胞24小时后,经MTT法检测细胞吸光度值,并计算细胞存活率。采用Agilent单标表达谱基因芯片对差异表达基因进行了筛选,并应用实时定量PCR技术随机选择了五个差异基因验证基因芯片结果的可靠性。结果MTT实验结果显示,杀草强剂量在1～100-g/ml时,Nthy-ori-3-1细胞的形态以及增殖同样无显著的变化,表明在此剂量下,杀草强对Nthy-ori-3-1细胞的影响与细胞毒性无关。基因芯片筛选结果显示,有90个基因的表达发生了显著的变化,其中55个基因的表达上调,35个基因表达下调；差异基因共影响了45个信号通路,其中大部分与肿瘤发生发展有关。实时定量PCR技术验证差异基因表达,AGT、CA11基因显著上调；MUC12、RAPH1、POLR1A基因显著下调,与基因芯片结果一致。结论杀草强剂量在1-100μg/ml时,其对Nthy-ori-3-1细胞的影响与细胞毒性无关。AGT、CAll、MUC12基因在甲状腺肿瘤的发生发展中起到很重要的作用；RAPH1、POLR1A基因能够影响细胞的内部结构,包括蛋白质的合成、修饰和代谢以及细胞骨架系统的装配,实时定量PCR验证结果与基因芯片结果一致。
[Abstract]:Objective thyroid gland is the largest endocrine gland in human body. Thyroid hormone secreted by thyroid gland plays an important role in growth, development and metabolism of human body. Thyroid disrupters have a significant effect on the thyroid gland of human and animals, which can interfere with thyroid function, induce autoimmune thyroid diseases, and have potential thyroid tumor effects. Pesticides account for a large proportion of known thyroid disruptions. As a non-selective herbicide, it has been proved to interfere with the synthesis of thyroid hormones and lead to thyroid hyperplasia and thyroid neoplasms. In this study, the human thyroid cell line Nthy-ori-3-1 cells were selected as subjects, and the mechanism related to the thyroid neoplasms induced by chlorpropoxide was explored at the gene level at the dose excluding cytotoxicity. Methods four dose groups of 0 渭 g / ml and 渭 g/ml.10pg/ml.100 渭 g/ml were designed. Human thyroid cell line Nthy-ori-3-1 cells were cultured on the culture medium containing the corresponding test material for 24 hours. The cell absorbance was measured by MTT method and the cell survival rate was calculated. The differentially expressed genes were screened by Agilent single standard gene microarray, and five differentially expressed genes were randomly selected by real-time quantitative PCR to verify the reliability of the results. Results the results of MTT test showed that the morphology and proliferation of Nthy-ori-3-1 cells had no significant change at the dose of 1~100-g/ml, which indicated that the effect of Xizuoqiang on Nthy-ori-3-1 cells was not related to the cytotoxicity. The results of gene chip screening showed that there were significant changes in the expression of 90 genes, of which 55 genes were up-regulated and 35 genes were down-regulated. Most of them are related to tumorigenesis and development. The differential gene expression was confirmed by real-time quantitative PCR, the AGT,CA11 gene was up-regulated and the MUC12,RAPH1,POLR1A gene was down-regulated, which was consistent with the results of gene chip. Conclusion the cytotoxicity of Nthy-ori-3-1 cells is not related to its effect on Nthy-ori-3-1 cells at a dose of 1-100 渭 g/ml. AGT,CAll,MUC12 gene plays an important role in the development of thyroid neoplasms, and RAPH1,POLR1A gene can affect the internal structure of the cells, including the synthesis of proteins. The results of modification and metabolism, assembly of cytoskeleton system, and real-time quantitative PCR were consistent with those of gene chip.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R114
本文编号：2274652
[Abstract]:Objective thyroid gland is the largest endocrine gland in human body. Thyroid hormone secreted by thyroid gland plays an important role in growth, development and metabolism of human body. Thyroid disrupters have a significant effect on the thyroid gland of human and animals, which can interfere with thyroid function, induce autoimmune thyroid diseases, and have potential thyroid tumor effects. Pesticides account for a large proportion of known thyroid disruptions. As a non-selective herbicide, it has been proved to interfere with the synthesis of thyroid hormones and lead to thyroid hyperplasia and thyroid neoplasms. In this study, the human thyroid cell line Nthy-ori-3-1 cells were selected as subjects, and the mechanism related to the thyroid neoplasms induced by chlorpropoxide was explored at the gene level at the dose excluding cytotoxicity. Methods four dose groups of 0 渭 g / ml and 渭 g/ml.10pg/ml.100 渭 g/ml were designed. Human thyroid cell line Nthy-ori-3-1 cells were cultured on the culture medium containing the corresponding test material for 24 hours. The cell absorbance was measured by MTT method and the cell survival rate was calculated. The differentially expressed genes were screened by Agilent single standard gene microarray, and five differentially expressed genes were randomly selected by real-time quantitative PCR to verify the reliability of the results. Results the results of MTT test showed that the morphology and proliferation of Nthy-ori-3-1 cells had no significant change at the dose of 1~100-g/ml, which indicated that the effect of Xizuoqiang on Nthy-ori-3-1 cells was not related to the cytotoxicity. The results of gene chip screening showed that there were significant changes in the expression of 90 genes, of which 55 genes were up-regulated and 35 genes were down-regulated. Most of them are related to tumorigenesis and development. The differential gene expression was confirmed by real-time quantitative PCR, the AGT,CA11 gene was up-regulated and the MUC12,RAPH1,POLR1A gene was down-regulated, which was consistent with the results of gene chip. Conclusion the cytotoxicity of Nthy-ori-3-1 cells is not related to its effect on Nthy-ori-3-1 cells at a dose of 1-100 渭 g/ml. AGT,CAll,MUC12 gene plays an important role in the development of thyroid neoplasms, and RAPH1,POLR1A gene can affect the internal structure of the cells, including the synthesis of proteins. The results of modification and metabolism, assembly of cytoskeleton system, and real-time quantitative PCR were consistent with those of gene chip.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R114
【参考文献】
相关期刊论文 前1条
1 张在香;脱碘酶[J];卫生研究;1998年05期
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