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

肝细胞癌患者抗PCNA抗体的研究。

• 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

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.用人肝癌细胞(PLC/PRF/5)、喉癌细胞(Hep-2)和地鼠肾细胞(BHK)三种培养基细胞，对160例肝细胞癌(I组)患者血清进行抗核抗体(ANA)间接免疫荧光检测。以66例非肝癌患者血清(II组)、62例慢性肝病患者血清(III组)和235例正常人血清(IV组)为对照。在HEP-2或PLC/PRF/5中检测到ANA，而在BHK中未检测到ANA的发生率为：I组36例(22.5%)，II组19例(27.5%)，Ⅲ组12例(19.4%)，Ⅳ组仅1例(0.4%)。抗HEP-2阳性而抗BHK阴性的抗核抗体效价I组和II组高于III组和IV组。用间接免疫荧光…法进一步检测了2例与HEP-2和PLC/PRF/5核反应的肝细胞癌患者血清更多的是使用其他4种转化细胞和4种未转化细胞。我们观察到，在这两种血清中，仅与转化细胞反应，与SLE患者血清中的反应不同。提示肝细胞癌患者血清中存在仅与转化细胞核反应的抗核抗体。我们需要了解这种肿瘤特异性抗体是否与增殖细胞核抗原抗体发生交叉反应。用亲和层析法纯化增殖细胞核抗原。用小牛胸腺作为抗原源。从抗增殖细胞核抗原阳性的狼疮血清中提取免疫球蛋白G组分，用亲和层析柱与兔肾提取液偶联，吸附其他抗可提取核抗原抗体。将抗增殖细胞核抗原抗体与配基凝胶偶联，制备成亲和层析柱。小牛胸腺丙酮粉用PBS匀浆，上清液用水透析，沉淀后作为粗品。将抗原通过抗增殖细胞核抗原偶联柱，洗脱部分即为增殖细胞核抗原。纯化的抗原相对分子质量为29kd，在免疫印迹和酶免疫分析中均仅与抗增殖细胞核抗原血清反应。较少

项目成果

今井明彦,清沢研道他: 肝臓. 27. 1539-1545 (1986)

Akihiko Imai、Kendo Kiyosawa 等：肝脏。27. 1539-1545 (1986)

今井明彦,清沢研道他: 肝臓. 28. 383-384 (1987)

Akihiko Imai、Kendo Kiyosawa 等：肝脏。28. 383-384 (1987)

清沢研道他: "培養B型肝癌細胞内hepatitis B nuclear antigen(HBNA)の細胞内Kinetics(消化器病学の進歩)" 日本医学館, P70-72 (1986)

Kendo Kiyosawa 等：“培养的 B 型肝癌细胞中乙型肝炎核抗原 (HBNA) 的细胞内动力学（胃肠病学进展）”日本医学博物馆，P70-72 (1986)

Imai H, Kiyosawa K, Furuta S.: "Diversity of fluorescent antinuclear antibodies in the reaction with various substrates in sera of patients with hepatocellular carcinoma." Jpn J Clinicalimmunology. 10. 580-585 (1987)

Imai H、Kiyosawa K、Furuta S.：“荧光抗核抗体与肝细胞癌患者血清中各种底物反应的多样性。”

Kiyosawa K et al: J.Gastroenterol Hepatol. 1. 371-378 (1986)

Kiyosawa K 等人：J.Gastroenterol Hepatol。