Significance of E2-EPF in esophageal cancer

1)基因表达谱利用寡核苷酸芯片分析了12例食管鳞癌患者手术切除的24例癌组织和正常组织标本中12,600个转录序列对应的总RNA表达水平。使用基因表达比率的等级聚类分析将12名患者分为两组。这项分析表明，结果中存在显著的差异。2)免疫印迹法检测食道癌组织和KYSE50组织中E2-EPF的表达，并用抗E2-EPF多克隆抗体AB4504、J8和J9进行免疫印迹。在使用AB4504的食道癌样本中，KYSE 50中不存在24 kDa的单手。而在KYSE-50和食道癌标本中用J8扩增出一条48 kDa的条带。3)免疫组织化学：应用抗E2-EPF多克隆抗体AP2120b对食管鳞癌石蜡包埋标本中的E2-EPF蛋白进行二步显影。E_2-EPF主要集中在癌细胞的细胞核表面。癌组织中E2-EPF蛋白的表达模式与下列变量中的任何一个相关：TNM分类、分期、侵犯淋巴管或血管、壁内转移或分化程度。

1)Gene expression profilingUsing oligonucleotide microarrays, we analyzed total RNA expression levels corresponding to 12,600 transcript sequences in 24 paired cancerous and normal tissue operative specimens from 12 patients with esophageal squamous cell carcinoma. Hierarchical clustering analysis using gene expression ratios divided the 12 patients into two groups. This analysis showed significant difierenoes in the outcomes. The expression of E2-EPF was up-regulated in the poor prognostic cluster.2)ImmunoblottingEsophageal cancer samples and KYSE 50 were analyzed by SDS-PAGE, followed by immunoblotting using an anti-E2-EPF polyclonal antibody of ab4504,J8,and J9. A single hand of 24kDa was present in KYSE 50, not in esophageal cancer samples by using ab4504. But a single band of 48kDa was present in KYSE 50 and esophageal cancer samples by using J8. These result suggested that a double quantity body of E2-EPF might be present in esophageal cancer.3)ImmunohistochemistryUsing paraffin-embedded specimens from patients with esophageal squamous cell carcinoma, E2-EPF prontein was detected using an anti-E2-EPF polyclonal antibody of AP2120b using a dextrau polymer conjugate two step visualization system. E2-EPF was enriched on the surface of nuclei in cancer cells. E2-EPF protein expression patterns within cancerous lesions were nat associated with any of the following variables : TNM classification, stage, invasion into lymphatic or blood vessels, intramural metastasis, or bistologic grades of differentiation.