兰州大学机构库 >生命科学学院
2,4-二氯酚诱导HL7702和MEFs细胞死亡的机制研究
Alternative TitleOn mechanism of 2,4-dichlorophenol-induced cell death in HL7702 and MEFs cells
张小宁
Thesis Advisor张迎梅 ; 黄德军
2014-12-06
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword2,4-二氯酚 细胞死亡 凋亡 自噬 坏死 氧化应激 未折叠蛋白反应 内质网应激 DNA甲基化
Abstract本研究以正常组织来源的两株细胞系HL7702和MEFs为模型,探讨2,4-DCP诱导细胞死亡的方式及其机理,以期为氯酚类化合物提供充分的毒理学资料,为其中毒机理和毒性预警提供参考。结果显示: 1. 2,4-DCP (0.50,0.75和1.00 mM)暴露24 h后可以明显抑制HL7702和MEFs细胞的生长,降低细胞活力,使细胞发生皱缩、变圆、胞浆空泡化等形态学改变。 2. 2,4-DCP暴露后导致HL7702和MEFs细胞发生凋亡、坏死和自噬性细胞死亡。 3. 2,4-DCP暴露诱导了活性氧(Reactive oxygen species, ROS)的爆发,导致细胞发生氧化应激。HMOX1和Nrf2 mRNA表达的升高以及细胞核内Nrf2蛋白表达的升高,表明HMOX1/Nrf2信号通路的激活参与了2,4-DCP诱导的氧化应答反应。 4. 转录组水平研究结果表明,2,4-DCP主要影响了细胞的固醇/胆固醇合成、内质网应激反应以及固醇/胆固醇代谢过程等与内质网功能密切相关的生物学途径,提示2,4-DCP主要通过影响内质网的功能体现其毒性作用。内质网应激相关基因Bip和CHOP在mRNA水平和蛋白水平的激活表明2,4-DCP可以诱导内质网应激的发生。进一步研究发现2,4-DCP暴露可激活IRE1α、ATF6、eIF2α三条未折叠蛋白反应,诱发内质网应激,并且eIF2α的去磷酸化介导了2,4-DCP诱导的内质网应激途径的细胞凋亡。 5. 利用甲基化DNA免疫共沉淀测序法在全基因组水平上分析2,4-DCP对HL7702细胞DNA甲基化的影响。结果显示,2,4-DCP暴露后干扰了DNA甲基化动态平衡。
Other AbstractIn the present study, human liver 7702 cells (HL7702) and mouse embryonic fibroblasts (MEFs) derived from normal tissue were used as in vitro models to study 2,4-DCP-induced cell death and its potential mechanisms in order to supply more adequate toxicity data and provide a reference for toxic mechanisms and toxicity warning of CPs. The results showed that: 1. 2,4-DCP (0.50, 0.75 and 1.00 mM) could inhibit the cell growth, reduce cell viability and induce significant morphological changes (including cellular shrinkage, cell rounding and cytoplasmic vacuolization) in HL7702 and MEFs. 2. 2,4-DCP could induce apoptosis, necrosis and autophagy in HL7702 and MEFs. 3. 2,4-DCP induced overproduction of reactive oxygen species (ROS) resulting in oxidative stress. The up-regulations of HMOX1 and Nrf2 mRNA expression and increase of Nrf2 protein in the nucleus demonstrated that the HMOX1/Nrf2 signaling pathway was involved in 2,4-DCP-induced oxidant responses. 4. The results from transcriptome level displayed that 2,4-DCP mainly affected endoplasmic reticulum (ER) function-related biological pathways including sterol/ cholesterol biosynthetic and metabolic process, response to ER stress, implying that 2,4-DCP cytotoxicity is closely related to ER dysfunction. We found that 2,4-DCP dramatically increased the expressions of Bip and CHOP both at mRNA and protein levels. Since induction of Bip and CHOP represented the activation of the ER stress, we further examined which of the three branches in ER stress were activated by 2,4-DCP. The results revealed that 2,4-DCP induced all three branches (IRE1α, ATF6 and eIF2α) of ER stress. In addition, 2,4-DCP induces ER stress-mediated apoptosis via eIF2α dephosphorylation. 5. We investigated the effect of 2,4-DCP (0.50 mM) on global DNA methylation with methylated DNA immunoprecipitation sequencing (MeDIP-seq) in HL7702 cells. The results showed that 2,4-DCP significantly disturbed methylation balance.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222094
Collection生命科学学院
Recommended Citation
GB/T 7714
张小宁. 2,4-二氯酚诱导HL7702和MEFs细胞死亡的机制研究[D]. 兰州. 兰州大学,2014.
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