摘要
目的以人甲状腺Nthy-ori-3-1细胞为受试对象,通过差异性表达谱芯片分析,探索杀草强致人甲状腺肿瘤的相关机制。方法以1-100μg/m L杀草强处理Nthy-ori-3-1细胞24 h后,用MTT法检测其对细胞增殖的影响。以100μg/m L杀草强处理细胞24 h后,做基因表达谱分析,并用GO（Gene Ontology）分析和pathway分析芯片结果,用实时定量PCR验证芯片结果。结果 MTT结果显示,所有检测浓度杀草强对Nthy-ori-3-1细胞增殖均无显著影响。芯片结果显示,有90个基因表达显著变化,55个上调,35个下调;GO分析显示,43个基因与生物过程相关（37个上调,6个下调）,42个与分子功能相关（37个上调,5个下调）,44个与细胞组分相关（38个上调,6个下调）。Pathway结果显示差异基因共影响45条信号通路,其中10条与肿瘤发生发展密切相关。实时定量PCR验证差异基因表达与芯片结果一致。wnt5b、arnt2和bmp2基因在多条肿瘤相关通路中均有显著变化。结论杀草强可能通过多信号通路导致甲状腺肿瘤,其中wnt5b、arnt2和bmp2等基因可能是其主要靶基因。
Objective To explore the possible mechanism of amitrole causing thyroid tumor in Nthy-ori-3-1 cell by differential expression microarray analysis. Methods After the Nthy-ori-3-1cells were treated with 1 - 100 g / m L amitrole for 24 h,and the effect of amitrole on the proliferation of the cells was detected by MTT assay. Then cells were treated with 100 g / m L amitrole for 24 h,and the differential expression microarray was tested. The microarray results was analyzed by GO analysis and pathway analysis. The microarray results were verified by real-time quantitative PCR. Results MTT results showed that amitole had no significant effect on the proliferation of Nthy-ori-3-1 cells.Microarray results showed that 90（ 55 up-regulated,35 down regulated） genes were significantly changed. GO analysis showed that 43（ 37 up-regulated,6 down-regulated） of the 90 changed genes were related to biological processes,and 42（ 37 up-regulated,5down-regulated） were related to molecular function,and 44（ 38 up-regulated,6 downregulated） were related to cell components. Pathway results showed that 44 signalingpathways were influenced by the differentially expressed genes,and 10 of them were closely related to tumor. The qRT-PCR results were consistent with microarray results.wnt5 b,arnt2 and bmp2 genes were significantly related with multiple tumor-associated pathways. Conclusion Amitrole may cause thyroid tumor by multiple signaling pathways,and bmp2,arnt2 and wnt5 b may beits major target genes.
出处
《卫生研究》
CAS
CSCD
北大核心
2016年第4期558-562,共5页
Journal of Hygiene Research
基金
云南省自然科学基金面上项目(No.2013FZ062)
