热疗对2’,3’-环核苷酸3’-磷酸二酯酶表达和C6胶质瘤细胞侵袭作用的影响  

Influence of hyperthermia in expression of 2',3'- cyclic nucleotide 3'-phosphodiesterase and invasion effect of C6 glioma cells

作　　者：王健[1] 张田[1] 贾永森[2] 赵喜庆[3] 秦丽娟[1] 

机构地区：[1]河北联合大学基础医学院生理学教研室,河北唐山063000 [2]河北联合大学中医学院,河北唐山063000 [3]河北联合大学附属唐山市人民医院神经外科,河北唐山063000

出　　处：《吉林大学学报（医学版）》2014年第2期248-252,I0001,共6页Journal of Jilin University：Medicine Edition

基　　金：河北省卫生厅科研基金资助课题(20100465;20110165;20120144);河北联合大学培育基金资助课题(SP201310)

摘　　要：目的:探讨热疗与2’,3’-环核苷酸3’-磷酸二酯酶(CNP)表达的关系,阐明热疗降低胶质瘤侵袭性的作用及其可能机制。方法:将对数生长期C6胶质瘤细胞随机分为对照组(37℃孵育)和热疗组(42℃孵箱内孵育10、30、60、120、180和240min)。各组细胞均行细胞爬片,采用免疫组织化学法检测各组细胞中热休克蛋白70(HSP70)的表达水平;Western blotting法检测各组细胞中CNP蛋白的表达水平;于Transwell上室构建胶质瘤侵袭模型,经热疗后采用结晶紫染色法观察透过Transwell下室面的胶质瘤细胞数。结果:与对照组比较,热疗组C6细胞经热疗后HSP70表达水平增加,于热疗30、60、120和180min时HSP70表达水平高对于照组(P<0.05或P<0.01),且于热疗30min时达高峰(P<0.01)。热疗组C6细胞中CNP蛋白表达水平亦于热疗后开始增加,热疗60、120和180min时CNP蛋白表达水平均高于对照组(P<0.05或P<0.01),并于热疗120min时达高峰(P<0.01)。热疗后透过Transwell的胶质瘤细胞数减少,细胞数于热疗120min时达到最低水平(P<0.01)。结论:热疗可能是通过增加HSP70的表达而使CNP蛋白的表达增多,增多的CNP又通过某种机制降低了胶质瘤的侵袭性。Objective To explore the relationship betwen hyperthermia and expression of 2＇, 3＇-cyclic nucleotide 3 ＇-phosphodiesterase （CNP）, and to clarify the effect of hyperthermia on decreasing the invasiveness of glioma cells and its possible mechanism. Methods C6 glioma cells of logarithmic phase were randomly divided into control group （37℃ incubation） and hyperthermia group （42℃ incubation for 10, 30, 60, 120, 180, and 240 min）. Immunohistochemical method was used to detect the expression levels of heat shock protein 70 （HSP70） in glioma cells in various groups; the expression levels of CNP protein were detected by Western blotting method; the cells in each group on the Transwell were used to build glioma invasion model. The number of glioma cells under the Transwell after hypertherimia was observed by crystal videt staining. Results Compared with control group, the HSP70 protein expression levels in C6 cells were increased after hyperthermia and reached the peak at 30 rain （P〈 0.01） and then gradually decreased, which was higher than those in control group after 30, 60, 120, and 180 rain hyperthermia（P〈0.05 or P〈0.01）. The expression levels of CNP protein in C6 cells began to increase after hyperthermia and reached the peak at 120 rain （P〈0.01） and then gradually decreased. The expression levels in CNP protein in C6 cells after 60, 120, and 180 rain hyperthermia were higher than those in control group（P〈0.05 or P〈0.01）. After hyperthermia, the number of glioma cells passing Transwell membrane reduced and reached the lowest at 120 min （P〈0.01）. Conclusion Hyperthermia may increase the expression level of CNP protein by increasing the expression of HSP70, and the increasing expression of CNP induces the invasion of glioma cells through some sort of mechanism.

关 键 词：热疗 热休克蛋白70 2’ 3’-环核苷酸3'-磷酸二酯酶 胶质瘤 侵袭性 

分 类 号：R74[医药卫生—神经病学与精神病学]