EFFECT OF HEAT TREATMENT OF TUMOR CELL ON LYMPHOKINE ACTIVATED KILLER CELL CYTOTOXICITY

肿瘤细胞热处理对淋巴细胞因子激活的杀伤细胞细胞毒性的影响

• 批准号: 06670501
• 负责人: NAKADA Tetsuya
• 金额: $ 0.9万
• 依托单位: THE JIKEI 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: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670501/
• 关键词: LAK cell; LAK activity; Hyperthermia; Tumor immunology; Cancer

The aim of this study is to demonstrate the effect of heat treatment of tumor cells on the cytotoxicity of these cells by lymphokine activated killer cell (LAK cell). Human hepatocellular carcinoma cell line, Huh-7 was used as target cell, and to evaluate both the cell proliferation of tumor cells and LAK activity, a new assay system was established using Alamar blue. At first, tumor cells were incubated at 41.0ﾟC,43.0ﾟC or 45.0ﾟC,then the proliferation rates of these treated cells were measured. Secondly, LAK cells and/or tumor cells were heated before LAK assay and the heat effect of these cells on LAK activity were evaluated. Finally, LAK assays were performed before or after heat treatment, then the total cytotoxicity induced by LAK cells and heat treatment were compared.Results :1. Tumor cells heated at 41.0ﾟC for 1-6 hours or at 43.0ﾟC or 45.0ﾟC for 1 hour showed the same proliferative activities as that of tumor cells without heat. On the other hand, heat treatment at 43.0ﾟC or 45.0ﾟC for 3-6 hours completely inhibited tumor growth.2. Heat treatment of tumor cells enhanced LAK activity by non-treated LAK cells.However, heat-treated LAK cells failed to show cytotoxicity against both heat-treated and not treated tumor cells.3. Heat treatment of tumor cells before LAK assay induced remarkable cytotoxic effect. However, when tumor cells were heated after LAK assay, the total cytotoxicity did not increase. Simultaneous LAK assay with heat showed little cytotoxicity against tumor cells.

本研究旨在观察热处理对淋巴因子激活的杀伤细胞（LAK细胞）杀伤肿瘤细胞的影响。以人肝癌细胞系Huh-7为靶细胞，采用阿尔玛蓝建立了一种新的检测体系，用于评价肿瘤细胞的细胞增殖和LAK活性。首先将肿瘤细胞置于41.0 ℃、43.0 ℃和45.0 ℃孵育，然后测定这些处理的细胞的增殖率。其次，在LAK活性测定前，对LAK细胞和/或肿瘤细胞进行加热，并评价这些细胞对LAK活性的热效应。最后，分别在热处理前后进行LAK细胞活性测定，比较热处理前后LAK细胞的总杀伤活性。肿瘤细胞在41.0 ℃加热1-6小时或43.0 ℃或45.0 ℃加热1小时，其增殖活性与未加热的肿瘤细胞相同。另一方面，43.0 ℃或45.0 ℃热处理3-6小时可完全抑制肿瘤生长.肿瘤细胞热处理可增强LAK细胞的杀伤活性，但热处理的LAK细胞对热处理和未处理的肿瘤细胞均无杀伤活性.在LAK试验前对肿瘤细胞进行热处理，可产生明显的细胞毒效应。然而，当肿瘤细胞在LAK试验后加热时，总的细胞毒性并没有增加。热诱导的LAK细胞对肿瘤细胞的杀伤作用很小。