冷冻联合5-氟尿嘧啶对G422胶质瘤细胞凋亡的影响  

Effect of Cryotherapy and 5-Fluorouracil on Apoptosis of G422 Glioma Cells

作　　者：王辉[1] 涂汉军[1] 秦军[1] 李新建[1] 黄宽明[1] 周章明[1] 王伦长[1] 

机构地区：[1]郧阳医学院附属太和医院神经外科,湖北十堰442000

出　　处：《癌症》2004年第4期412-415,共4页Chinese Journal of Cancer

基　　金：湖北省科技攻关项目(编号:2001AA307B16)~~

摘　　要：背景与目的:冷冻与5-氟尿嘧啶(5-FU)均能诱导胶质瘤细胞凋亡,二者联合应用能否产生协同效应尚未知。本实验旨在观察冷冻联合5-FU对胶质瘤细胞凋亡的影响,并对其内在的分子机制进行探讨。方法:(1)建立G422胶质瘤动物模型,随机分组,分别施以冷冻(-180℃,90s)、化疗(5-FU18mg/kg)、冻化疗;(2)TUNEL法观察不同时间点各组细胞凋亡的变化;(3)免疫组化方法检测冷冻后HSP90α及p53蛋白的表达。结果:冷冻与5-FU均能引起肿瘤细胞凋亡,二者联合应用后细胞凋亡率明显增加;联合组于冻化疗后6、12、24、48、72h细胞凋亡数(每0.0625mm2范围内的细胞个数)分别为30.6±11.7、86.4±21.5、128.1±4.1、237.0±30.1、72.8±23.0,均明显多于冷冻组和化疗组。冷冻中心区无HSP90α和p53的阳性表达,冷冻后周边区表达比对照组高,尤以冻后48h最为明显犤分别为(73.1±9.3)%vs(60.6±9.9)%、(35.6±6.6)%vs(13.7±6.5)%(P<0.01)犦。冻后HSP90α与p53的表达呈正相关(r=0.3610,P<0.01)。结论:冷冻联合5-FU能够明显提高G422胶质瘤的细胞凋亡率,其发生机制可能与冷冻后HSP90α与p53表达调节有关;冷冻与化疗的联合应用较单纯冷冻或化疗作用恶性胶质瘤效果更为明显。BACKGROUND & OBJECTIVE: It was unknown whether the combinatio n of cryotherapy and 5- fluorouracil (5- FU) could produce a synergistic effect, though both of them could induce apoptosis of glioma cells. The aim of this stu dy was to observe the effect of cryotherapy and 5- fluorouracil on apoptosis of G422 glioma cells and to investigate the potential mechanism. METHODS:(1)Experi mental mice models with G422 glioma were established. Cryotherapy (- 180℃ ,90s ),chemotherapy (5- FU, 18 mg/kg), and cryochemotherapy were performed on them,r espectively.(2)TUNEL was used to determine the apoptosis of glioma cells in each group at different time points.(3)The expression of the p53 and the HSP90α ge ne of the frozen glioma were analyzed by immunohistochemical staining. RESULTS: Cryotherapy or 5- FU induced apoptosis of G422 glioma cells, while the combinat ion of cryotherapy and 5- FU remarkably increased the apoptosis rate. The numbe rs of apoptotic cells of combination group were 30.6± 11.7, 86.4± 21.5, 128.1 ± 4.1, 237.0± 30.1, 72.8± 23.0 at 6h, 12h, 24h, 48h, 72h after cryochemothera py, respectively, which were significantly higher than those of other groups. Th ere was no expression of HSP90α and p53 in the center of freezing area. The p5 3 and the HSP90α proteins increased obviously in the periphery area especially at 48h after freezing compared with control group [(73.1± 9.3)% vs (60.6± 9 .9)% , (35.6± 6.6)% vs (13.7± 6.5)% ](P< 0.01). The expression of p53 was positively correlated with expression of HSP90α in frozen glioma (r=0.3610, P< 0.01). CONCLUSION: Combination of cryotherapy with 5- FU could enhance the apo ptosis of G422 glioma cells presumably through modulating the HSP90α and p53 e xpression pattern. Cryochemotherapy showed better effect than that of cryotherap y or chemotherapy alone.

关 键 词：冷冻治疗 5-氟尿嘧啶 G422胶质瘤 细胞凋亡 5-FU 肿瘤细胞 

分 类 号：R73-3[医药卫生—肿瘤]