"Biological effects of soluble recombinant CD40 ligand and interferon-γ on human renal cancer cell lines in vitro and in vivo."

“可溶性重组 CD40 配体和干扰素-γ 对人肾癌细胞系的体外和体内生物学效应。”

• 批准号: 10671497
• 负责人: FUJII Tsunehiro
• 金额: $ 0.83万
• 依托单位: 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671497/
• 关键词: CD40; renal cell cancer; SCID mouse; interferon-γ; renal cell canser; SCID mouse; インターフェロンγ; SCID マウス

1. Surface expression of CD40 on human renal cancer cell line.Immunofluorescence studies were performed by flow cytometric analysis. CD40 was expressed on various human renal cancer cell lines, 769-P, 786-O, A-498 and A-704.2. Induction of CD40 expression by interferon-γ in vitro.Incubation with interferon-g significantly induced surface expression on human renal cancer cell lines.3. Antitumor effects of soluble recombinant CD40 lingand (srCD40L) and interferon-γ on human renal cancer cell lines in vitro.Incubation with srCD40L significantly inhibited the proliferation of 769-P and 786-O as determined by the proliferation assay with optimal inhibition of thymidine incorporation. Apoptotic studies by flow cytometry revealed an induction of apoptosis in these two cell lines. Additional inhibitory effect by co-incubation with interfron-γ significantly enhanced this inhibitory effects.4. Antitumor effects of srCD40L treatment in SCID mice bearing human human renal cancer cell lines.Effects of srCD40L treatment on survival in human renal cancer cell-bearing SCID mice were determined. All mice received 20 μL of anti-asialo GMI (Wako Chemicals, Osaka) by intravenous injection (IV) 1 day before tumor transfer to remove host natural killer cells. 769-P cells (5 x l0ィイD16ィエD1) were then administered by IV. SCID recipients then received either l0 μg of srCD40L or controll every other day for 20 days for total of 10 injections starting at day 3. Tumor bearing mice were then monitored for tumor development and progression. Treatment with srCD40L significantly improved the survival of tumor-bearing mice (p<0.05).We are preparing a manuscript from these data for Japanese Journal for Cancer Research.

1.人肾癌细胞系CD40的表面表达，流式细胞仪免疫荧光检测。CD40在各种人肾癌细胞系769-P、786-O、A-498和A-704上表达。干扰素-γ体外诱导人肾癌细胞系CD40表达干扰素-g可显著诱导人肾癌细胞系CD40表达.可溶性重组CD 40配体（srCD 40 L）和γ-干扰素（IFN-γ）对人肾癌细胞株的体外抗肿瘤作用研究表明，srCD 40 L对769-P和786-O细胞的增殖有明显的抑制作用，对胸腺嘧啶核苷（TdR）掺入的抑制作用最佳。通过流式细胞术的凋亡研究揭示了在这两个细胞系中的凋亡诱导。与干扰素-γ共孵育的额外抑制作用显著增强了这种抑制作用. srCD40L处理对携带人肾癌细胞系的SCID小鼠的抗肿瘤作用测定srCD40L处理对携带人肾癌细胞的SCID小鼠的存活率的作用。所有小鼠在肿瘤转移前1天通过静脉内注射（IV）接受20 μ L抗脱唾液酸GMI（和子Chemicals，Osaka）以去除宿主自然杀伤细胞。然后通过IV给予769-P细胞（5 x 10 μ g/ml D16 μ g/ml D1）。然后SCID受体从第3天开始每隔一天接受10 μ g的srCD40L或对照，持续20天，总共10次注射。然后监测荷瘤小鼠的肿瘤发展和进展。srCD40L治疗显著提高了荷瘤小鼠的存活率（p <0.05）。我们正在为日本癌症研究杂志准备这些数据的手稿。