The Study on anti-PCNA antibody in patient with hepatocellular carcinoma.

肝细胞癌患者抗PCNA抗体的研究。

基本信息

批准号：
61570334
负责人：
KIYOSAWA Kendo
金额：
$ 1.41万
依托单位：
Shinshu University School Of Medicine.
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61570334/
关键词：
proliferating cell nuclear antigen (PCNA)anti-PCNA antinuclear antibody PCNA 核抗原肝癌

项目摘要

1. Sera from 160 patients with hepatocellular carcinoma(group I) were examined for anti-nuclear antibodies(ANA) indirect immunofluorescence using 3 types of cultured as substraye : human hepatoma cell(PLC/PRF/5), laryngeal carcinoma cell(HEp-2) and baby hamster kidney cell (BHK). Sixty six sera from patients with non-hepatic cancers (group II), 62 sera from patients with chronic liver disease (group III) and 235 sera of normal subjects (group IV) were tested as controls. The incidences of ANA which was detected in HEp-2 or pLC/PRF/5 but not in BHK were as follows, 36 patients(22.5%) in group I, 19 patients(27.5%) in group II, 12 patients (19.4%) in group III and only 1 case(0.4%) in group IV. Reciprocal titers of ANA that were positive against HEp-2 but negative against BHK in group I and II were higher than in group III and IV. Two Two sera of patients with hepatocellular carcinoma reacted with nuclei of HEp-2 and PLC/PRF/5 were investigated further for ANA by indirected immunofluores … More cence using other 4 types of transformed cells and 4 types of non-transformed cells. We observed that in these two sera reacted only with transformed cell differed from that in sera of patients with SLE. It was suggested that the ANA reacting only with nuclei of transformd cells was present in sera of patients with hepatocellular carcinoma. We need weather this tumor specific antibody cross react with antibody to proliferating cell nuclear antigen(PCNA).2. Purification of PCNA was performed by affinity chromatography. Calf thymus was used as a source of antigen. IgG fraction was obtained from anti-PCNA positive lupus serum and absorbed other antibodies to extractable nuclear antigens by affinity column coupled with rabbit kidney extract. Anti-PCNA IgG antibody was conjugated with ligand gels to make PCNA affinity coloumn. Calf thymus acetone powder was homogenized with PBS and the supernatant was dialysed against water, then the precipitate was used as crude PCNA. The antigen was passed over the anti-PCNA coupled column and the eluted fraction was taken as PCNA. This purified antigen was a 29 kd in molecular weight andreacted only with anti-PCNA serum in both immunoblotting and enzyme immunoassay. Less

1。检查了160例肝细胞癌患者（I组）的血清抗核抗体（ANA）间接免疫荧光，使用3种培养为副施法的类型：人类肝瘤细胞（PLC/PRF/5），喉癌细胞（Hep-2）和Baby Hamsney Cell（Hepf/prf/prf/prf/5）从非肝脏癌症患者（II组），62例慢性肝病患者（III组）和235例正常受试者（IV组）的62例血清（第IV组）（IV组）的66名血清（第四组）。在HEP-2或PLC/PRF/5中检测到的ANA的发明如下，I组为36例（22.5％），II组的19例患者（27.5％），III组的12例（19.4％），III组中只有1例（19.4％），IV组中只有1例（0.4％）。对HEP-2呈阳性的ANA的倒数滴度，但在I组和II组中对BHK负的效率高于III和IV组。通过间接免疫荧光素进一步研究了ANA的两种两种与HEP-2和PLC/PRF/PLF/PRF/5的肝细胞癌患者的血清……使用其他4种转化的细胞和4种非转化细胞。我们观察到，在这两种血清中，仅与转化的细胞反应不同于SLE患者的血清。有人提出，仅在肝细胞癌患者的血清中存在ANA与核转化细胞反应。我们需要天气这种肿瘤特异性抗体交叉与抗体反应以增殖细胞核抗原（PCNA）.2。 PCNA的纯化是通过亲和色谱进行的。小腿胸腺被用作抗原的来源。从抗PCNA阳性狼疮血清获得IgG馏分，并通过亲和度柱与兔肾提取物吸收其他可提取的核抗原的抗体。将抗PCNA IgG抗体与配体凝胶偶联以使PCNA亲和力Colourn。小牛胸腺丙酮粉与PBS匀浆，并将上清液透析在水上，然后将沉淀物用作粗pcna。抗原通过抗PCNA耦合色谱柱，并以PCNA为pCNA。这种纯化的抗原是分子量的29 kD，仅在免疫印迹和酶免疫测定中都用抗PCNA血清进行反应。较少的