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Screening and identification of peritoneal metastasis-related genes of gastric adenocarcinoma using a cDNA microarray

机译:利用cDNA微阵列筛选和鉴定胃腺癌腹膜转移相关基因

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With the aim of identifying peritoneal metastasis-related genes in gastric cancer, we performed a broad analysis of differential gene expression between the parental cell line GC9811 and its highly metastatic peritoneal counterpart, cell line GC9811-P. Two fluorescent cDNA probes, labeled with Cy3 and Cy5 dyes, were prepared from GC9811 and GC9811-P mRNA samples by the reverse transcription method. The two color probes were then mixed and hybridized to a cDNA chip constructed with double-dots from 11,901 human genes; this was scanned at two wavelengths. The experiment was repeated twice. In GC9811-P cells, 218 genes were upregulated and 30 genes were downregulated compared with the parental cell lines. Some selected genes were confirmed by RT-PCR and Western blot; we found that S100A4 and CTNNB1 were upregulated and PTEN was downregulated in GC9811-P cells. Identification of these differentially expressed genes could contribute to disclose the molecular mechanisms involved and provide new targets for therapeutic intervention to avoid peritoneal dissemination of gastric adenocarcinoma.
机译:为了鉴定胃癌中腹膜转移相关基因,我们对亲代细胞系GC9811及其高度转移性腹膜对应细胞系GC9811-P之间的差异基因表达进行了广泛的分析。通过逆转录方法从GC9811和GC9811-P mRNA样品中制备了两个用Cy3和Cy5染料标记的荧光cDNA探针。然后将两种颜色的探针混合并杂交到一个cDNA芯片上,该芯片由来自11,901个人类基因的双点构成;在两个波长扫描。重复实验两次。与亲本细胞系相比,GC9811-P细胞中有218个基因上调,有30个基因下调。通过RT-PCR和Western印迹证实了一些选择的基因;我们发现在GC9811-P细胞中S100A4和CTNNB1被上调,而PTEN被下调。这些差异表达基因的鉴定可能有助于揭示所涉及的分子机制,并为治疗干预提供新的靶标,以避免腹膜扩散性胃腺癌。

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