TRAIL诱导骨肉瘤HOS-8603细胞凋亡与线粒体跨膜电位关系的研究  

Study on the relationship between HOS-8603 cell apoptosis induced by TRAIL and the cell mitochondrial transmembrane potential .

作　　者：喻爱喜[1] 周志刚[1] 张仲宁[1] 祝少博[1] 

机构地区：[1]武汉大学中南医院显微骨科,湖北武汉430071

出　　处：《现代肿瘤医学》2005年第5期586-588,共3页Journal of Modern Oncology

基　　金：湖北省自然科学研究基金资助项目(编号2003ABA163)

摘　　要：目的研究TRAIL诱导HOS-8603细胞凋亡与线粒体跨膜电位(ΔΨm)关系。方法MTT法测定TRAIL对HOS-8603细胞的抑制率;将HOS-8603细胞分别用TRAIL、TRAIL+1.0μg/ml环孢菌素A(CsA)处理,然后用透射电镜观察HOS-8603细胞形态学变化;用流式细胞术分别测定亚G1期细胞百分率及PI和Rh123双染色后细胞的ΔΨm。结果MTT还原试验表明:作用24小时后,2.5μg/mlTRAIL的抑制率为29%;5μg/mlTRAIL的抑制率为83%。HOS-8603细胞经TRAIL作用后,透射电镜下观察到典型的凋亡形态特征;随着TRAIL作用时间增加,HOS-8603细胞凋亡数增加和ΔΨm降低(F=33.831P<0.01),两者呈直线相关。CsA不仅能抑制TRAIL诱导HOS-8603细胞ΔΨm降低,而且能减少凋亡细胞数的增加(t=7.103,P<0.01)。结论TRAIL诱导HOS-8603细胞凋亡的线粒体依赖途径是通过使线粒体膜通透性转运孔开放,ΔΨm降低来实现;CsA能部分抑制这些效应。Objective To explore the relationship between HOS-8603 cell apoptosis induced by TNF- related apoptosis - inducing ligand （ TRAIL ） and the cell mitochondrial transmembrane potential （ΔΨm） . Methods TRAIL - mediated cytotoxicity by MTT assay in HOS - 8603 osteosarcoma cell line; HOS - 8603 cells were treated with TRAIL and TRAIL plus 1.01μg/ ml CsA respectively. Morphological changes were observed under light microscope and transmission electron microscope. The percentages of subG1 cells and ΔΨm of cells doublely stained with PI and Rh123 were assayed by flow cytometry. Results HOS -8603 cells were treated for 24 hours with different concentration of TRAIL and the inhibitive rates of TRAIL were evaluated with MTT assay , the inhibitive rates of 500μg/ml,1μg/ml,2μg/ml,2.5μg/ml and 5μg/ml ,TRAIL inducing HOS -8603 osteosareoma cell line was 1.6 % ,9.89 % ,16 % ,29 % and more than 83 % respectively. After treatment with TRAIL , Hos -8603 cells appeared the classical apoptotie morphology. The percentages of subG, cells were increased , while the ΔΨm reduced （ F = 33. 831 P 〈 0.01 ） and there was a linear correlation between them. The increment of subG, cell percentages and decre- ment of ΔΨmm induced by TRAIL were partly inhibited by CsA （t = 7. 103 P 〈 0.01 ） . Conclusion TRAIL can partly induce HOS -8603 cells apoptosis through opening the mitoehondrial permeability transition pore and reducing ΔΨm , and this effect could be partly inhibited by CsA.

关 键 词：TRAIL 细胞凋亡 HOS-8603细胞系 环孢菌素 线粒体跨膜电位 

分 类 号：R735.2[医药卫生—肿瘤]