Significance of E2-EPF in gastric cancer

E2-EPF在胃癌中的意义

• 批准号: 16591355
• 负责人: TORIUMI Yasuo
• 金额: $ 1.98万
• 依托单位: The Jikei University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591355/
• 关键词: E2-EPF; Gastric Cancer; Ubiquitin

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.

1）RT-PCR检测胃癌细胞系MKN 1、MKN 45、EC 10、GT 3 TkB中E2-EPF基因的表达。2）免疫印迹法采用SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）和抗E2-EPF多克隆抗体ab 4504、AP 2120 a进行免疫印迹分析，检测E2-EPF mRNA在胃癌细胞系MKN 7、MKN 45、IM 95和食管癌细胞系KYSE 50中的表达。用ab 4504对KYSE 50和IM 95进行电泳，结果显示一条分子量为24 kDa的条带。而ab 4505在MKN 7和MKN 45中均出现一条48 kDa的条带。用AP 2120 a检测各样品中与E2-EPF偶联的泛素链的不同条带。免疫组化应用抗E2-EPF多克隆抗体AP 2120 b，采用葡聚糖聚合物二步法显色系统，检测胃癌组织中E2-EPF蛋白的表达。E2-EPF在癌细胞核表面富集。E2-EPF蛋白在癌性病变中的表达模式与以下变量无关：TNM分类、分期、淋巴管或血管浸润、壁内转移或组织学分化程度。