细胞周期蛋白激酶抑制剂flavopiridol对尤文肉瘤耐药细胞增殖抑制的实验研究  

Inhibitory effect of cyclin-dependent kinase inhibitor flavopiridol on the proliferation of WE-68/ADR cells in vitro and in vivo

作　　者：李旭[1] 李岩 程世孝[1] 李雷明[1] 孟凡贺[1] 朱悦[1] 范广宇[1] 

机构地区：[1]中国医科大学附属第一医院骨科,辽宁沈阳110001

出　　处：《现代肿瘤医学》2013年第1期17-20,共4页Journal of Modern Oncology

基　　金：国家自然科学基金面上项目(编号:30973021)

摘　　要：目的:探讨细胞周期蛋白激酶抑制剂(flavopiridol)在体内外抑制尤文肉瘤阿霉素耐药细胞株WE-68/ADR增殖的作用及其机制。方法:四甲基偶氮唑蓝法(MTT法)测定细胞增殖抑制率。流式细胞计数法测量flavopiridol给药后WE-68/ADR细胞周期的变化及凋亡率。免疫印迹法(Western-blot)检测细胞中Bax、Bcl-2、p53及活化型多聚ADP核糖多聚酶(PARP-85)蛋白的表达。皮下注射WE-68/ADR细胞建立荷瘤裸鼠模型,17flavopiridol腹腔内给药,测量肿瘤体积及裸鼠体重变化,计算肿瘤抑制率。结果:Flavopiridol可抑制尤文肉瘤阿霉素耐药细胞株WE-68/ADR的细胞增殖,其效果呈时间及浓度相关性(P<0.05)。Flavopiridol给药后,耐药细胞株WE-68/ADR的细胞凋亡比明显高于对照组(P<0.05)。同时,耐药细胞株中的Bax、p53及PARP-85的表达增加,而Bcl-2的表达被下调。Flavopiridol可有效抑制荷瘤小鼠体内耐药细胞株的生长。结论:细胞周期蛋白激酶抑制剂flavopiridol可在体内外有效抑制尤文肉瘤阿霉素耐药细胞株的增殖,其机制可能与p53介导的线粒体凋亡信号传导途径有关。Objective ：To investigate the inhibitory effect of cyclin- dependent kinase inhibitor （flavopiridol） on the proliferation of adriamycin - resistant Ewing~ sarcoma cells, WE - 68/ADR and its underlying mechanism in vitro and in vivo. Methods ：The proliferation of WE - 68/ADR cells was determined by using MTr method. Cell cycle progres- sion and apoptosis ratio were determined by flow cytometry. The expression of Bax,p53, bcl - 2, cleaved - PARP were de- tected by western blot. The xenograft model was successfully established via injecting WE - 68/ADR cells subcutane- ously in nude mice. Mice were monitored for changes in the tumor volume：and body weight after flavopifidol was injec- ted intraperitoneally, Results：After exposure to flavopiridol,the growth of WE -68/ADR cells was inhibited in a time - and dose - dependent manner （ P 〈 0.05 ）. Apoptosis rate of WE - 68/ADR was higher than that of control group （ P 〈 0.05 ）. The expression of bcl - 2 was down - regulated, and the expression of p53, Bax and cleaved - PARP were up - regulated. The inhibitory effect of flavopiridol on ADR clones of Ewing＇s sarcoma in the treatment group was ob- served compared with the control group in vivo. Conclusion ： Flavopiridol can inhibit the growth of drug - resistant EFTs cells in vitro and in vivo ,which might be mediated by p53 -dependent mitochondrial apoptosis pathway.

关 键 词：细胞周期 尤文肉瘤 P53 凋亡 

分 类 号：R73-362[医药卫生—肿瘤] R738.1[医药卫生—临床医学]