Borkamo Erling Dahl, Dahl Olav, Bruland Ove, Fluge Øystein
Section of Oncology, Institute of Medicine, University of Bergen, Bergen, Norway.
Int J Hyperthermia. 2009 Feb;25(1):41-6. doi: 10.1080/02656730802397955.
To analyze effects of hyperthermia on immune cells within tumors.
Subcutaneously implanted rat gliomas were treated with water-bath hyperthermia for one hour at day zero (HT) (intratumoral temperature of 43 degrees C), low-dose metronomic cyclophosphamide (CTX) 35 mg/kg three times a week for two weeks, both HT and CTX (CTX-HT(0)), or saline. Tumors harvested at day 1, 7, 14 and 21 were analyzed for changes in gene expression by microarrays, focusing on genes expressed in immune cells. Microarray analyses and real-time RT-PCR were previously performed on tumors harvested day zero at 0, 45, 90 and 180 minutes after end of treatment. Gene expression kinetics of selected genes were analyzed in all tumors by quantitative real-time RT-PCR (qPCR) to validate the results of the microarrays.
Previous microarray analyses have indicated a suppression of gene expression in immune cells within tumors by hyperthermia the first three hours after treatment. qPCR analyses of CD14, CD36, Klrd1 (CD94), Tlr2 and Lrp1 confirmed these results. Global mRNA screen revealed no hyperthermia-induced changes in gene expression of immune cells at day 1, 7, 14 and 21. qPCR analyses of CD14, CD36, Klrd1 (CD94), Tlr2 and Lrp1 confirmed the results of the microarrays, i.e. none of these were differentially expressed after the first day, with the exception of Lrp1 which displayed elevated mRNA levels.
The suppression in gene expression of a range of immune cells within tumors treated with hyperthermia at 43 degrees C is a transient phenomenon.
分析热疗对肿瘤内免疫细胞的影响。
将皮下植入的大鼠胶质瘤在第0天用水浴热疗1小时（HT）（瘤内温度为43摄氏度），低剂量节拍性环磷酰胺（CTX）35mg/kg，每周3次，共2周，热疗联合CTX（CTX-HT(0)），或生理盐水。在第1、7、14和21天收获肿瘤，通过微阵列分析基因表达变化，重点关注免疫细胞中表达的基因。之前已对治疗结束后0、45、90和180分钟收获的第0天肿瘤进行了微阵列分析和实时逆转录聚合酶链反应。通过定量实时逆转录聚合酶链反应（qPCR）分析所有肿瘤中选定基因的基因表达动力学，以验证微阵列的结果。
之前的微阵列分析表明，热疗后最初三小时内肿瘤内免疫细胞的基因表达受到抑制。对CD14、CD36、Klrd1（CD94）、Tlr2和Lrp1的qPCR分析证实了这些结果。整体mRNA筛选显示，在第1、7、14和21天，热疗未引起免疫细胞基因表达的变化。对CD14、CD36、Klrd1（CD94）、Tlr2和Lrp1的qPCR分析证实了微阵列的结果，即除Lrp1 mRNA水平升高外，第一天后这些基因均无差异表达。
43摄氏度热疗治疗的肿瘤内一系列免疫细胞的基因表达抑制是一种短暂现象。
