顺铂增加人脑胶质瘤细胞U251对TRAIL敏感性的实验研究  

Senciflvity enhancement of U251 cells to TRAIL by cisplatin

作　　者：孙晓柏[1] 温培娥[1] 陈剑[2] 任霞[1] 张恒兰[1] 苏恩裕[1] 唐天华[1] 任海全[1] 姜国胜[1] 

机构地区：[1]山东省医学科学院基础医学研究所,山东省现代医用药物与技术重点实验室,山东省医药卫生肿瘤免疫与中药免疫重点实验室,济南250062 [2]济宁市第一人民医院神经外二科

出　　处：《国际肿瘤学杂志》2010年第5期386-389,396,共5页Journal of International Oncology

基　　金：山东省博士后科研项目择优资助基金项目（200602024）

摘　　要：目的研究顺铂在增加人脑胶质瘤细胞U251对肿瘤坏死因子相关凋亡诱导配体（TRAIL）敏感性的作用，并探讨其分子机制。方法通过倒置荧光显微镜下观察重组腺病毒载体携带的绿色荧光蛋白的表达确定最适感染复数MOI；运用MTF法检测细胞增殖活性；倒置荧光显微镜下观察Hoechst 33342染色细胞凋亡形态；PI染色后通过流式细胞术检测诱导凋亡作用，RT-PCR法检测凋亡相关基因。结果感染后TRAIL基因的表达明显上调。顺铂增敏TRAIL组与单独组相比，有显著抑制细胞增殖作用（P〈0．05），Hoechst 33342染色观察有明显的核固缩，核碎裂。流式细胞术检测细胞有凋亡峰的出现，增敏组与单独组有显著性差异（P〈0．05）。RT-PCR检测到TRAIL、DR5、caspase-3基因上调，survivin基因下调。DR4表达无明显变化。结论顺铂可以增加人脑胶质瘤细胞U251对TRAIL的敏感性，其分子机制可能与DR5、caspase-3基因上调，survivin基因下调有关。Objective To evaluate the positive effects of cisplatin on sensitivity of human glioma U251 to tumor necrosis factor-related apoptosis inducing ligand and to investigate the potential mechanism. Methods The expression of green fluorescent protein （GFP）in U251 which was transfected with pAdxsi-GFP-TRAIL was observed by inverted fluorescent microscope （ × 400）and to ascertain the MOI. The proliferation inhibition was studied by MTT method. Morphological change was detected through inverted florescent microscope and the Hoechst33342 staining assay was used to verify whether cell apoptosis could be induced or not. The cell apoptosis was also analyzed by flow cytometry with propidium iodide staining. Semi-quantitative RT-PCR was introduced to detect the mRNA expression of apoptosis related gene. Results The expression of TRAIL mRNA was signifi- cantly upregulated after transfection. Compared with treatment group of cisplatin and TRAIL alone, the proliferation of U251 was significantly inhibited in the cisplatin sensitizing TRAIL group（ P 〈 0. 05 ）. Nuclear shrinkage and pyknosis fragmentation were observed by Hoechst 33342 staining assay ; Apoptotic peak was detected from the results of flow cytometry and there were significant differences between the sensitizing group and the other two groups （ P 〈 O. 05 ） ; Moreover, the relatively high expression of TRAIL, DRS, caspase3 and down- regulated survivin genes were also observed. There was no significant changes in DR4 expression. Conclusion Cisplatin could extremely enhance the sensitivity of U251 cells to TRAIL And the potential mechanism may related to the increase of TRAIL, DRS, caspase3 genes while the reduction of surivivin gene.

关 键 词：神经胶质瘤 TNF相关凋亡诱导配体 腺病毒科 顺铂 

分 类 号：R739.4[医药卫生—肿瘤]