肝癌凋亡细胞相关抗原的初步研究  

Preliminary studies of apoptotic cell associated antigens on hepatoma

作　　者：马运国[1] 王文亮[1] 

机构地区：[1]第四军医大学病理学教研室

出　　处：《中华肿瘤杂志》1998年第1期22-24,共3页Chinese Journal of Oncology

基　　金：国家自然科学基金

摘　　要：目的分析肝癌细胞凋亡反应中瞬时表达的某些相关抗原分子。方法利用消减免疫法制备抗肝癌凋亡细胞相关抗原的抗体，ＥＬＩＳＡ和免疫斑点法检测抗原抗体反应，免疫细胞化学法鉴定抗原的细胞内定位。结果该抗体与肝癌凋亡细胞相关抗原反应较强，与非凋亡的肝癌细胞抗原反应较弱，其抗原决定簇主要是蛋白质或多肽。该抗原主要表达于肝癌凋亡细胞的细胞膜、细胞浆及细胞核未见该抗原的表达。结论凋亡的肝癌细胞有一定的表型，可用来检测细胞的凋亡反应；凋亡过程中有新基因的表达，可用该抗体筛选肝癌凋亡细胞新表达的基因。Objective To analyse antigens transiently expressed on hepatoma cells during apoptotic response. Methods Hepatoma HCC9204 cells were treated with 6% ethanol for 6 hr to induce apoptosis. Apoptotic HCC9204 cells were isolated and homogenized. After centrifugation at 14 000 rpm, the supernatant was dialyzed against water with molecular weight cutoff at 12 000. The dialysate was used as hepatoma apoptotic cell associated antigen (HACAA). Subtractive immunization technique was used to prepare polyclonal antibodies against apoptotic cell associated antigens on hepatoma. The antibody response to HACAA was determined by ELISA and immunoblot assays. Localization of HACAA on hepatoma apoptotic cells was demonstrated immunocytochemically. Results This polyclonal antibody reacted more strongly with HACAA than with antigen likewise prepared from non apoptotic hepatoma cell. The antigenic determinants were sensitive to trypsin and mainly located on the cytoplasmic membrane.Conclusion Epitopes are expressed on hepatoma apoptotic cells which can be used to examine an apoptotic response. The antibody reported in this paper is useful to screen novel apoptosis associated genes.

关 键 词：肝肿瘤 细胞凋亡 抗原 消减免疫法法 

分 类 号：R735.7[医药卫生—肿瘤]