VP-16诱导细胞凋亡及其细胞核基质中热休克蛋白表达的研究  

THE STUDIES ON EXPRESSION OF HEAT SHOCK PROTEIN IN APOPTOTIC CELL AND NUCLEAR MATRIX INDUCED BY VP-16

作　　者：金梅林[1] 丁明孝[2] 云径平[3] 张萍[3] 陈约翰[3] 邱殷庆[3] 

机构地区：[1]华中农业大学畜牧兽医学院,武汉430070 [2]北京大学生命科学院,北京100871 [3]香港中文大学医学院

出　　处：《解剖学报》2002年第1期88-92,共5页Acta Anatomica Sinica

摘　　要：目的　研究VP 16诱导HL 6 0细胞凋亡的变化特点以及凋亡细胞及核基质中热休克蛋白 (HSP)表达的变化。　方法　琼脂糖凝胶电泳观察凋亡细胞基因组DNA断裂 ;TUNEL法观察凋亡细胞的形态学变化 ;用免疫印迹方法显示凋亡细胞核基质蛋白、细胞总蛋白中HSP表达的差异。　结果　VP 16作用HL 6 0细胞 0 5h出现早期凋亡特征。作用 4h可见明显的DNA梯状条带。与未凋亡的细胞比较 ,凋亡细胞核基质蛋白中HSP70和HSC70表达明显上调 ;VP 16作用 2、3及 4h细胞总蛋白中HSP70的表达随时间延长略显增加 ;诱导 2 2h比诱导 4h时去除VP 16后孵至 2 2h凋亡细胞总蛋白HSP70表达量增强了 3 4倍。　结论　 1 HL 6 0细胞早期凋亡形态出现在DNA梯状条带形成之前。 2 凋亡细胞核基质中HSP70、HSC70的表达明显增加 ,经 2、3及 4h作用的细胞总蛋白中HSP70表达差异不显著。这提示细胞凋亡后HSP70的活性位点主要位于核基质上。 3 VP 16作用细胞时间增长至2 2h 。Objective To study the characteristic morphological changes of HL 60 cell apoptosis induced by VP 16 and the expression of heat shock protein in apoptotic nuclear matrix protein (NMP) in comparision with the cell total protein(CTP). Methods Apoptosis was determined by DNA fragmentation assessed by agrose gel electrophresis. The morphological change of apoptosis was observed by TUNEL method. The influence of apoptotisis on NMP and CTP was shown by Western blotting. Results The early apoptotic cells were observed after VP 16(40μg/L) treatment for 0 5?h. The ladder of DNA fragments of approximately 200bp was observed by VP 16 treatment for 4h. The expression of HSP70 and HSC70 increased dramatically in the nuclear matrix, comparing with normal control. And there was a few alteration in the CTP. The expression of CTP of apoptosis significantly decreased in the HL 60 with VP 16 treatment for 22?h. However, after remove VP 16 at 4?h and incubation for 22?h continually, the expression of HSP70 was about 3 4 fold greater than that in the cells of un removed VP 16.Conclusion 1 The change of apoptotic morphology displayed earlier than DNA ladder. 2 The expression of the HSP70 and HSC70 was significantly up regulated in apoptotic nuclear matrix proteins after 4h treatment with VP 16. There was no difference in HSP expression in the CTP between treated(2,3,4?h) and untreated HL 60 cells. It suggests that the site of apoptotic action of HSP70 may be localized in nuclear matrix other than cytoplasm. 3 Incubation with VP 16 for 22?h may lead to the protective role of heat shock protein disappearing. [

关 键 词：细胞凋亡 HL-60细胞 核基质蛋白 细胞总蛋白 热休克蛋白 

分 类 号：R73－36[医药卫生—肿瘤]