应用基因芯片技术研究非毒性结节性甲状腺肿基因表达的初步研究

摘要

Objective To screen the related genes for nontoxic nodular goiter (NTNG) and to explore the pathogenic mechanism of NTNG. Methods Total RNA was extracted and purified from thyroid tissues of NTNG cases and normal cases. The cDNAs retro-tran-scribed from RNA were labeled with Cy5 and Cy3 fluorescence as probes. The probes were hybridized with gene chip. The chip was scanned and the acquired imaging was analyzed. Parts of these genes were analyzed and identified by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR). Results Of the 48 genes with altered expressions, 36 genes were up-regulated and 12 genes were down-regulated. The results identified by FQ-PCR consistent with those of gene chip. Conclusion Marked difference exists in gene expression in thyroid tissues of NTNG patients and normal thyroid tissues, showing the complex alterations of gene expression underlying the development of NTNG. The obtained genes are mainly related with extracellular matrix, cytokine, signal transduction and so on. The identification of specific genes is helpful for early diagnosis and treatment of NTNG.%目的：筛查非毒性结节性甲状腺肿（NTNG）相关基因，探讨NTNG发生机制。方法分别抽提NTNG患者甲状腺组织及正常甲状腺组织总RNA，逆转录合成相应以Cy5和Cy3标记的探针与基因表达谱芯片进行杂交，扫描芯片荧光信号图像，用对扫描图像进行数字化处理和分析。利用实时荧光定量PCR技术对筛选出的部分表达差异明显的基因进行验证。结果筛选出表达差异明显的基因48个，其中NTNG组中表达上调的基因36条，表达下调的基因12条。荧光定量PCR结果与基因芯片一致。结论 NTNG患者甲状腺组织与正常甲状腺组织之间存在明显的基因表达差异，为NTNG患者提供相当数量的与细胞外基质、细胞因子、受体信号转细胞信号传递等相关的差异表达基因，为NTNG早期诊断和治疗提供了线索。