Relation between Hyperthermia Reduce Glioma Invasiveness and TNFR Affinity
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摘要: 目的 探讨热疗降低胶质瘤侵袭性的作用与肿瘤坏死因子受体(tumor necrosis factor receptor，TNFR)亲和力的关系。方法免疫组织化学法观察肿瘤坏死因子受体在胶质瘤组织中的分布；免疫荧光法检测肿瘤坏死因子受体在热疗后胶质瘤组织中的分布；放射配基受体结合分析法测定胶质瘤细胞经热疗联合重组人肿瘤坏死因子(recombinant human tumor necrosis factor，rhTNF）作用后，胶质瘤细胞中rhTNF结合肿瘤坏死因子受体水平的改变；通过荧光分光光度计分析Caspase3/7的活性来检测胶质瘤细胞的凋亡水平；结晶紫染色法测定热疗后胶质瘤侵袭性的变化。结果TNFR存在于胶质瘤细胞和胶质瘤血管内皮细胞，但以TNFR1分布于胶质瘤细胞为主；热疗后的胶质瘤组织中TNFR1的表达明显高于TNFR2，且于热疗后的120 min时TNFR1的表达达到高峰；胶质瘤细胞经热疗联合rhTNF作用后，rhTNF与TNFR1的结合率和Caspase3/7的活性呈上升趋势，两者均于120 min时达高峰后下降，与此同时，胶质瘤侵袭性降至最低水平。结论 热疗后，rhTNF可能是通过增加胶质瘤细胞TNFR1的表达并与之结合，并增强了rhTNF与TNFR1的亲和力，进而引起胶质瘤细胞凋亡导致胶质瘤侵袭性降低的。Abstract: Objective To assess the relation between hyperthermia reduce glioma invasiveness and TNFR affinity. Methods Using immunohistochemical method and immunofluorescence method，respectively，to detect the expression and distribution of TNFR in tumor tissue.Radioligand binding assay of receptors method monitor the receptor binding rate of rhTNF after combination hyperthermia with rhTNF in C6 cells.Detection of C6 cells apoptosis by fluorescence spectrophotometric analyzed activity of Caspase3/7.Crystal violet staining method was used to detect the glioma invasiveness. Results In glioma tissue，TNFR2 expressed slightly on the tumor capillaries and tumor cells，TNFR1 expression mainly on the tumor cells.Hyperthermia 120 min group TNFR1 immunoreactivity had a significant increase.The receptor binding rate of rhTNF and activity of Caspase3/7 were significant increased and they both reached the peak at 120 min(compared with control group，P<0.01)，then decreased.At the same time，glioma invasiveness reached the minimum. Conclusion Hyperthermia increases TNFR1 expression and receptor binding rate of rhTNF further causes tumor cells apoptosis.This mechanism may explain the effect of hyperthermia on reducing glioma invasiveness.
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Key words:
- Hyperthermia /
- Glioma invasiveness /
- Tumor necrosis factor receptor
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