胃癌淋巴结转移相关基因差异表达分析

Screening of related genes in lymphatic metastasis of gastric carcinoma

关键词:  基因芯片  胃肿瘤  淋巴结  转移相关基因  差异表达

Key words:  gene chip  gastric neoplasms  lymph nodes  related gene of metastasis  differential expression

摘要:

［目的］探讨胃癌淋巴结转移过程中相关基因群的表达和功能。［方法］从6例胃癌患者分别抽取胃癌和淋巴结转移组织的总RNA，采用逆转录的方法，制成cDNA链，并以两种荧光Cy5和Cy3标记后做探针，与含有14784条人类14KcDNA基因表达谱芯片进行杂交。以Agilent荧光扫描仪扫描芯片上两种荧光信号，进行计算机处理和分析，筛选出胃癌淋巴结转移组织中差异表达的基因，并用RT—PCR实验对部分差异表达基因进行验证。［结果］ 在14784条基因中，胃癌原发灶与淋巴结转移组织间存在差异表达的基因共80条，其中59条基因表达显著上调, 21条基因表达显著下调。经RT-PCR验证，差异表达趋势与基因芯片技术检测的结果一致。［结论］胃癌组织和淋巴结转移组织之间存在多个差异表达的基因，胃癌淋巴结转移的发生与发展是多种相关基因表达失常所致。

Abstract:

［Objective］To search for the differentially expressed genes between lymphatic metastasis tissue and primary gastric carcinomas using gene expression profile chip.［Methods］Total RNA from gastric carcinoma and lymphatic metastasis tissue was extracted and purified. The cDNA was made by RT-PCR, and then labeled with Cy5 and Cy3 fluorescence as probes which were hybridized with 14784 human 14K cDNA gene expression profile chip. Subsequently, the signal images were scanned by Agilent fluorescence scanner and analyzed by gene Pix QuantArrayR. The gene expression profile chip screened the related genes in lymphatic metastasis of gastric carcinoma and made a further confirmation by using semi-quantity RT-PCR.［Results］Among the 14784 target genes, 80 genes were differentially expressed in metastatic lymph nodes, including 59 were up-regulated and 21down-regulated.As proved by RT-PCR, expressed tendency of difference is in accordance with result of gene chip.［Conclusions］There are many genes differentially expressed between lymphatic metastasis tissue and primary gastric carcinomas .This study demonstrated that many different kinds of genes are possibly involved in the initiation and progression of lymphatic metastasis of gastric carcinoma.