热疗降低胶质瘤侵袭性的作用与TNF-α和Cx-43的关系

Relation between hyperthermia reduce glioma invasiveness and changes of TNF-α and Cx-43

目的:探讨热疗诱发胶质瘤细胞释放的肿瘤坏死因子-α(TNF-α)对胶质瘤细胞侵袭性的影响机制。方法:热处理C6细胞后,应用放射免疫法动态监测培养液内TNF-α的含量;观察热处理C6细胞后的条件培养液(C6CM)对胶质瘤细胞内的丝裂素活化蛋白激酶(mitogen-activated protein kinase,MAPK)mRNA的转录水平、NF-κB含量及胶质瘤侵袭性的影响;利用免疫荧光技术检测C6CM对体外胶质瘤侵袭模型上连接蛋白Cx-43表达的影响。结果:热处理C6细胞后,培养液内TNF-α的含量增加,于120 min时达高峰。C6CM作用于体外胶质瘤侵袭模型后,胶质瘤细胞的MAPK mRNA转录水平及NF-κB含量增加,且均于第120 min时达最高水平。与此同时,体外胶质瘤细胞的连接蛋白Cx-43表达水平也呈相同的变化趋势。结论:热疗可诱发C6细胞释放TNF-α,其可能是通过增加胶质瘤细胞的MAPK mRNA转录水平和NF-κB的含量而导致胶质瘤细胞的连接蛋白Cx-43的表达增加,进而引起胶质瘤侵袭性降低的。

Objective：To assess the effect and mechanism of tumor necrosis factor-α（TNF-α） induced by hyperthermia from glioma cells to glioma invasiveness.Methods： We used radioimmunoassay to dynamically monitor contents of TNF-α in nutrient fluid for C6 cells after hyperthermia treatment.The effects of conditioned medium of C6（C6CM） treated by hyperthermiaon on levels of mitogen-activated protein kinase（MAPK） mRNA and contents of nuclear transcription factor kappaB（NF-κB） in glioma cells,as well as glioma invasiveness.By using immunofluorescence technique,the effects of C6CM on the expression of Cx-43 in glioma cells.Results： Contents of TNF-α in the nutrient fluid for C6 cells increased obviously after hyperthermia treatment and achieved the maximal level at 120 min.After adding C6CM in vitro glioma invasiveness model,levels of MAPK mRNA and contents of NF-κB were both increased,reached the maximum level at 120 min.And the expression of Cx-43 showed the same trend.Conclusion： Hyperthermia promoted C6 cells to release TNF-α.The TNF-α increased the expression of NF-κB and levels of MAPK mRNA in C6 cells,then enhanced expression of Cx-43,which decreased glioma invasiveness.