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

本研究旨在探讨肿瘤细胞热处理对淋巴因子激活的杀伤细胞(LAK细胞)杀伤作用的影响。以人肝细胞癌细胞系Huh-7为靶细胞，以阿拉马蓝为试剂，建立了检测肿瘤细胞增殖和LAK活性的新体系。首先将肿瘤细胞置于41.0ﾟC、43.0ﾟC和45.0ﾟC的环境中孵育，然后测定这些处理细胞的增殖率。其次，在LAK实验前对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。结果：1.肿瘤细胞在41.0ﾟC加热1~6小时，或在43.0ﾟC或45.0ﾟC加热1小时后，其增殖活性与未加热的肿瘤细胞相同。另一方面，43.0ﾟC或45.0ﾟC热处理3-6小时完全抑制肿瘤生长。肿瘤细胞的热处理增强了未处理的LAK细胞的LAK活性，但热处理的LAK细胞对热处理和未处理的肿瘤细胞均未表现出细胞毒作用。LAK检测前对肿瘤细胞进行热处理可诱导出明显的细胞毒效应。然而，LAK实验后加热肿瘤细胞，其总的细胞毒作用并没有增加。LAK与热法同时检测，对肿瘤细胞无明显杀伤作用。

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.