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为探讨预热对肿瘤细胞肿瘤坏死因子α(TNF-α)敏感性的影响及其机理,将白血病患者胸腔渗出细胞系(K562细胞)在40℃预热不同时间(0、30、60、90、120、150和180min),使细胞热应激蛋白70(HSP70)高表达;然后将预热60min细胞和未预热细胞分别暴露于50、100、200和400U/ml的TNF-α16小时,用MTT比色法检测细胞活力。保持TNF-α浓度为100U/ml不变,观察预热时间对细胞TNF-α敏感性的影响,用逆转录聚合酶链反应(RT-PCR)测定细胞HSP70的含量。结果:40℃预热后细胞的HSP70含量升高,在预热120min达到最高,以后缓慢下降;预热细胞对相同浓度TNF-α的敏感性明显低于未预热细胞;对于相同浓度的TNF-α,预热120min的细胞对TNF-α敏感性最低。结果提示,预热可以引起细胞HSP70表达增高,降低细胞对TNF-α的敏感性。
To explore the effect of warm-up on the sensitivity of tumor cells to tumor necrosis factor-alpha (TNF-alpha) and its mechanism, the leukemia patient’s thoracic exudate cell line (K562 cells) was preheated at 40[deg.]C for different time (0, 30, 60, 90, 120, 150, and 180 min), high expression of cellular heat stress protein 70 (HSP70); 60 min preheated cells and unheated cells exposed to 50, 100, 200, and 400 U/ml TNF-α for 16 hours, respectively The cell viability was measured by MTT colorimetry. Keeping the TNF-α concentration unchanged at 100 U/ml, observe the effect of preheating time on TNF-α sensitivity of cells, and determine the content of HSP70 in cells by reverse transcription polymerase chain reaction (RT-PCR). Results: The HSP70 content of cells preheated at 40°C increased, reached the highest at 120 min after warm-up, and then decreased slowly. The sensitivity of preheated cells to TNF-α at the same concentration was significantly lower than that of unheated cells; the same concentration of TNF -α, cells preheated for 120 min were least sensitive to TNF-α. The results suggest that preheating can increase the expression of HSP70 in cells and reduce the sensitivity of cells to TNF-α.