Identification of genes responsive to mild hyperthermia in human skin normal fibroblast Hs68 cells. Homo sapiens

Hyperthermia is widely used to treat patients with various cancers. Here, the effects of heat stress at 41°C for 30 min (mild hyperthermia) on the gene expression in Hs68 human skin normal fibroblast cells were investigated using an Affymetrix GeneChip system. The cells were treated with mild hyperthermia, followed by incubation for 0, 1, or 3 h at 37°C. No cell death was observed in the mild hyperthermia-treated cells. On the other hand, many genes that were differentially expressed by a factor 1.5 or greater were identified in the cells treated with the mild hyperthermia. Overall design: Hs68 human skin normal fibroblast cells were treated with heat stress (41°C for 30 min), followed by incubation for 0, 1, or 3 h at 37°C. Non-treated cells served as the control. Total RNA samples were prepared from the cells, and quality of the RNA was analyzed using a Bioanalyzer 2100. Gene expression was monitored by an Affymetrix GeneChip® system with a Human Genome U133-plus 2.0 array. Sample preparation for array hybridization was carried out as described in the manufacturer's instructions.

热疗（Hyperthermia）已被广泛应用于多种癌症的临床治疗。本研究采用Affymetrix GeneChip系统，探究了41℃热应激30分钟（即温和热疗，mild hyperthermia）对Hs68人皮肤正常成纤维细胞基因表达的影响。实验细胞经温和热疗处理后，于37℃下分别孵育0、1或3小时。经温和热疗处理的细胞未出现细胞死亡现象；而在该处理组细胞中，共鉴定出表达差异倍数达1.5倍及以上的大量差异基因。整体实验设计如下：对Hs68人皮肤正常成纤维细胞施加41℃热应激30分钟处理，随后于37℃下分别孵育0、1或3小时；以未经过处理的细胞作为对照组。从各组细胞中提取总RNA样品，并采用Bioanalyzer 2100对RNA的质量进行检测。采用搭载Human Genome U133-plus 2.0芯片的Affymetrix GeneChip®系统监测基因表达水平。芯片杂交所需的样品制备流程严格遵循生产商的说明书进行操作。