Analysis of thermotolerance mechanism in carcinoma thermotherapy by cDNA microarrays method

在肿瘤热疗治疗中，既有治愈的，也有不治愈的。人们认为癌细胞的热敏性和耐热性存在差异，但其根本因素尚未明确。利用cDNA微阵列技术和已确认的诱导基因分析了其热敏性、耐热性机制。肺成纤维细胞损伤DNA双链断裂修复和杂化scid细胞损伤与热敏性、耐热性和DNA双链断裂有关。Scid细胞具有放射敏感性，引入人类染色体的杂交Scid细胞恢复到正常的放射敏感性。我们研究了耐热性抑制剂维生素K2的作用。与杂交Scid细胞和对照Balb/c 3T3细胞相比，Scid细胞表现出较高的热敏性。我们在处理Scid细胞后观察到核内Y-H2AX聚焦。在30分钟后热疗处理后，我们逐渐降低并达到最大值。Y-H2AX病灶随热疗时间的延长而增加。将a Scid细胞与Hybrid Scid细胞和Balb/c 3T3细胞的Y-H2AX灶数进行比较，差异明显。核内Y-H2AX灶与热疗敏感性增高有相关性。在这里，我们发现维生素k2抑制热休克蛋白72的表达，但不影响热休克蛋白70的组成表达。维生素ink2使A549细胞和HaLa细胞对热处理诱导的细胞死亡敏感。这些发现提示细胞的热敏性和耐热性可能与DNA双链断裂的形成有关。

In carcinoma hyperthermia therapy, we have experienced carcinoma to be cured well and the carcinoma that are not cured. It is thought that there is a difference of thermosensibility and thermotolerance of cancer cell, but an essential factor is not yet clarified. We analyze thermosensibility, thermotolerance mechanism with cDNA microarrays method and an confirmed inductive gene. We demanded relation with thermosensibility, thermotolerance and DNA double strand breaks with lung fibroblast scid cells suffered a loss in DNA double strand breaks repair and Hybrid scid cells. A Scid cell is radiosensitive, and the Hybrid Scid cell which introduced the human chromosome8 into recovers to normal radiosensitive. We examined an effect of thermotolerance inhibitor Vitamin K2. Scid cells showed high thermosensibility in comparison with Hybrid Scid cells and a Balb/c 3T3 cells using control. We observed intranuclear Y-H2AX focus after hyperthermia processed a Scid cell. We gradually decreased with a maximum after hyperthermia disposal afterwards in 30 minutes. The Y-H2AX focus increased with hyperthermia treatment time. There was different clearly when it compared the number of the Y-H2AX focus of a Scid cells with Hybrid Scid cells and Balb/c 3T3 cells. A correlation was seen in increase and hyperthermia sensibility of intranuclear Y-H2AX focus. Here, we show that vitamin k2 inhibited the expression of heat-shock protein 72 but did not affect the constitutive expression of heat-shock protein 70. VitaminK2 sensitized A549 cells and HaLa cells to heat-treatment induced cell death. These finding suggest that thermosensitivity and thermotolerance of cells might be associated with DNA double-strand break formation.