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目的:结合激光微切割(LMD)和基因芯片技术筛选胃癌淋巴结转移相关基因,从中进一步确定在胃癌发生和转移过程中都有差异表达的基因。方法:采用LMD技术,从17例胃癌患者手术标本中获取高纯度的正常胃黏膜细胞及原发灶和相应淋巴结转移灶中的肿瘤细胞。用基因芯片比较其中2例原发灶与淋巴结转移灶中肿瘤细胞基因表达谱的差异,半定量RT-PCR方法验证芯片结果;并在另15份标本中,进一步确定在胃癌发生和转移过程中都有差异表达的基因。结果:发现49个基因的表达在胃癌原发灶肿瘤细胞中比淋巴结转移灶明显上调,37个基因的表达明显下调,mRNA水平的验证结果与芯片结果相符。同时,筛选出4个在胃癌发生和转移过程中都有差异表达的基因,其中3个基因(OPCML,RNASE1和YES1)的表达规律与肿瘤抑制基因相似,即正常黏膜中表达最高,肿瘤原发灶次之,淋巴结转移灶表达最低;而另一个基因(AKC1)的表达规律则与癌基因相似。结论:采用LMD和基因芯片技术,成功筛选出胃癌进展过程中的差异表达基因,为进一步研究胃癌发生和转移的机制奠定了良好的基础。
OBJECTIVE: To screen the genes related to lymph node metastasis of gastric cancer by laser microdissection (LMD) and gene chip technique, and to further identify the genes that are differentially expressed during the carcinogenesis and metastasis of gastric cancer. Methods: The normal gastric mucosa with high purity and the tumor cells in the primary tumor and the corresponding lymph node metastasis were obtained from the surgical specimens of 17 patients with gastric cancer using LMD technique. Gene chips were used to compare the gene expression profiles of tumor cells in two cases of primary and metastatic lymph nodes. Semi-quantitative RT-PCR was used to verify the chip results. In another 15 samples, There are differentially expressed genes. Results: The expression of 49 genes was found to be significantly up-regulated in lymph node metastasis and the expression of 37 genes was significantly down-regulated in the primary tumor of gastric cancer. The validation of mRNA level was consistent with that of the chip. At the same time, four genes that were differentially expressed during the carcinogenesis and metastasis of gastric cancer were screened. Among them, the expression pattern of three genes (OPCML, RNASE1 and YES1) was similar to the tumor suppressor gene, which was the highest expression in normal mucosa, Followed by the lowest level of lymph node metastasis, while the expression of the other gene (AKC1) was similar to oncogene. Conclusion: The differentially expressed genes during the progression of gastric cancer were successfully screened by LMD and gene chip technology, which laid a good foundation for further study on the mechanism of gastric cancer occurrence and metastasis.