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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 investigate the mechanism of herbicide-induced thyroid neoplasms by using differential expression profiling of human thyroid Nthy-ori-3-1 cells. Methods The Nthy-ori-3-1 cells were treated with 1 ~ 100μg / m L of herbicide for 24 h and their cell proliferation was measured by MTT assay. The cells were treated with 100μg / m L of herbicide for 24 hours, then the gene expression profiles were analyzed and the chip results were analyzed by GO (Gene Ontology) and pathway, and the chip results were verified by real-time quantitative PCR. Results The results of MTT assay showed that all concentrations tested showed no significant effect on the proliferation of Nthy-ori-3-1 cells. The results of the chip showed that 90 genes were significantly changed, 55 were up-regulated and 35 were down-regulated. GO analysis showed that 43 genes were related to biological processes (37 up-regulated and 6 down-regulated) and 42 were related to molecular function Up-regulated, 5 down-regulated) and 44 were related to cellular components (38 up-regulated, 6 down-regulated). Pathway results showed that a total of 45 differential signaling pathways were affected by differential genes, of which 10 were closely related to tumorigenesis. Real-time quantitative PCR verified the differential gene expression consistent with the chip results. The wnt5b, arnt2 and bmp2 genes were significantly altered in multiple tumor-associated pathways. Conclusion The strong killing of thyroid tumor may be caused by multiple signal pathways. The genes of wnt5b, arnt2 and bmp2 may be their main target genes.