遺伝子発現を利用した照射効果測定

利用基因表达测量辐照效果

• 批准号: 11670870
• 负责人: HIRANO Kazuya
• 金额: $ 2.24万
• 依托单位: TOKYO UNIVERSITY OF PHARMACY AND LIFE SCIENCE (2000)The University of Tokyo (1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670870/
• 关键词: MUC1; Radiation; Tumor anitigen; MUCl; 放射線; 腫瘍抗原; 放射線応答; ムチン; X線

The effect of X-irradiation on production of MUC1 was studied with human colon carcinoma HT-29 cells. Monoclonal antibody (mAb) MY.1E12 was used to identify MUC1. Immunomicroscopically, the percentage of MUC1 positive cell was 52% in 6 Gy irradiated cells at day 4, compared with 26% in unirradiated cells. By flow cytometric analysis, MUC1 on 6 Gy irradiated cells was elevated at day 1, reaching plateau by day 4. Westerm blot analysis showed both 500 kDa and 390 kDa bands corresponding to two polymorphic MUC1 alleles were increased after X-irradiation. The transcriptional activity of the MUC1 gene was analyzed by transient expression of MUC1-CAT reporter plasmid containing the 5'-flanking sequence of the MUC1 gene fused to the bacterial chloramphenicol acetyltransferase (CAT) gene. Elevation of CAT activity was observed in parallel with the expression of MUC1 after 6 Gy of X-irradiation. Moreover, to investigate in vivo effects, we developed a stable high-expression green fluorescent protein (GFP) transfectant of human colon carcinoma HT-29 cells. GFP-expressing cells (HT-29/GFP) were transplanted into the hind rimb of nude mice. After 40 days, tumor-bearing nude mice were irradiated of 0, 6, 10, 20, 30, 50 Gy. After 4 days, Westem blot analysis showed both 500 kDa and 390 kDa bands were increased of tumor derived HT-29/GFP cells after X-irradiation. These findings indicate that enhanced expression of MUC1 in irradiated HT-29 cells was due to upregulation of the MUC1 transcription.

用人结肠癌 HT-29 细胞研究了 X 射线照射对 MUC1 产生的影响。使用单克隆抗体 (mAb) MY.1E12 来鉴定 MUC1。免疫显微镜下，第4天经6 Gy照射的细胞中MUC1阳性细胞的百分比为52%，而未照射的细胞中MUC1阳性细胞的百分比为26%。通过流式细胞术分析，6 Gy 照射的细胞上的 MUC1 在第 1 天升高，在第 4 天达到平台。Western 印迹分析显示，对应于两个多态性 MUC1 等位基因的 500 kDa 和 390 kDa 条带在 X 射线照射后均增加。通过 MUC1-CAT 报告质粒的瞬时表达来分析 MUC1 基因的转录活性，该质粒含有与细菌氯霉素乙酰转移酶 (CAT) 基因融合的 MUC1 基因的 5' 侧翼序列。 6 Gy X 射线照射后，观察到 CAT 活性升高与 MUC1 表达平行。此外，为了研究体内效应，我们开发了一种稳定的高表达绿色荧光蛋白（GFP）的人结肠癌HT-29细胞转染子。将表达 GFP 的细胞 (HT-29/GFP) 移植到裸鼠的后肢。 40天后，对荷瘤裸鼠进行0、6、10、20、30、50Gy的照射。 4天后，Westem印迹分析显示，X射线照射后，肿瘤来源的HT-29/GFP细胞的500 kDa和390 kDa条带均增加。这些发现表明，受辐射的 HT-29 细胞中 MUC1 表达增强是由于 MUC1 转录上调。

项目成果

Yun Kang, Kazuya Hirano, Norio Suzuki, Atsushi Enomoto, Akinori Morita, Tatsuro Irimura, and Kazuo Sakai: "Increased expression after X-irradiation of MUC1 in cultured human colon carcinoma HT-29 cells."Jpn.J.Cancer Res.. 91. 324-330 (2000)

Yun Kang、Kazuya Hirano、Norio Suzuki、Atsushi Enomoto、Akinori Morita、Tatsuro Irimura 和 Kazuo Sakai：“培养的人结肠癌 HT-29 细胞中 MUC1 经 X 射线照射后表达增加。”Jpn.J.Cancer Res..

Y. Kang, K. Hirano, N. Suzuki, A. Enomoto, A. Morita, T. Irimura, and K. Sakai: "Increased expression after X-irradiation of MUCI in cultured human colon carcinoma HT-29 cells"Jpn. J. Cancer Res.. 91(in press). (2000)

Y. Kang、K. Hirano、N. Suzuki、A. Enomoto、A. Morita、T. Irimura 和 K. Sakai：“培养的人结肠癌 HT-29 细胞中 MUCI X 射线照射后表达增加”Jpn。

Y.Kang,K.Hirano,N.Suzuki et al.: "Increased expression after X-irradiation of MUCl in cultured human colon carcinoma HT-29 cells."Jpn.J.Cancer Res.. 91. 324-330 (2000)

Y.Kang，K.Hirano，N.Suzuki 等：“培养的人结肠癌 HT-29 细胞中 MUC1 X 射线照射后表达增加。”Jpn.J.Cancer Res.. 91. 324-330 (2000