Significance of E2-EPF in gastric cancer

1)RT-PCR检测胃癌细胞株（MKN1、MKN45、ECC10、GT3TkB） E2-EPF基因的表达。E2-EPF mRNA在各组中均有表达。2)免疫印迹法对胃癌细胞株（MKN7、MKN45、IM95）和食管癌细胞株（KYSE 50）进行SDS-PAGE分析，并用ab4504、AP2120a抗e2 - epf多克隆抗体进行免疫印迹。利用ab4504在KYSE 50和IM95中发现了一个24kDa的单带。而通过ab4505在MKN7和MKN45中存在单个48kDa带。用AP2120a对E2-EPF偶联的多泛素链各条带进行定向。提示胃癌组织中可能存在双量E2-EPF。3)免疫组织化学：利用胃癌患者石蜡包埋标本，采用葡聚糖聚合物偶联两步可视化系统，采用抗E2-EPF多克隆抗体AP2120b检测E2-EPF蛋白。E2-EPF在癌细胞细胞核表面富集。癌灶内E2-EPF蛋白表达模式与以下任何变量无关：TNM分类、分期、侵入淋巴或血管、壁内转移或组织学分化等级。

1)RT-PCRGastric carcinoma cell lines (MKN1,MKN45,ECC10,GT3TkB) were examined for gene expression of E2-EPF using RT-PCR. mRNA expression of E2-EPF was observed in all of them.2)ImmunoblottingGastric carcinoma cell lines (MKN7,MKN45,IM95) and esophageal carcinoma cell line (KYSE 50) were analyzed by SDS-PAGE, followed by immunoblotting using an anti-E2-EPF polyclonal antibody of ab4504,and AP2120a. A single band of 24kDa was present in KYSE 50 and IM95 by using ab4504. But a single band of 48kDa was present in MKN7 and MKN45 by using ab4505. Various bands of muriti-ubiquitin chain conjugated with E2-EPF were ditected in each sample by using AP2120a. These result suggested that a double quantity body of E2-EPF might be present in gastric cancer.3)ImmunohistochemistryUsing paraffin-embedded specimens from patients with gastric cancer, E2-EPF protein was detected using an anti-E2-EPF polyclonal antibody of AP2120b using a dextran 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 not associated with any of the following variables : TNM classification, stage, invasion into lymphatic or blood vessels, intramural metastasis, or histologic grades of differentiation.