(R)-Roscovitine诱导Hela细胞凋亡的分子机制  

Caspase-Dependent and-Independent Pathways for (R)-Roscovitine-Induced Apoptosis in Hela Cells

作　　者：张均平[1,2] 刘会杰[1,2] 杨洋[1,2] 许瑞安[1,2] 

机构地区：[1]华侨大学分子药物研究院,福建厦门361021 [2]华侨大学分子药物教育部工程研究中心,福建厦门361021

出　　处：《华侨大学学报（自然科学版）》2013年第4期422-428,共7页Journal of Huaqiao University(Natural Science)

基　　金：国家自然科学基金资助项目(81072578)

摘　　要：以Hela细胞为模型,探讨细胞周期蛋白依赖性激酶抑制剂(R)-roscovitine对Hela细胞凋亡诱导的可能途径,进一步了解其诱导肿瘤细胞凋亡的分子机制.结果显示:(R)-roscovitine明显抑制Hela细胞的增殖,在17.5～140.0μmol.L-1浓度作用12,24,48h后,Hela细胞的增殖抑制率分别为8.48%～33.76%,25.19%～86.41%和47.90%～88.08%;典型凋亡的DNA ladder被诱导,且凋亡率随药物浓度的增加而升高;经半胱天冬酶-3(caspase-3)特异性抑制剂Ac-DEVD-CHO或半胱天冬酶泛抑制剂Z-VAD-FMK处理后,(R)-roscovitine对Hela细胞凋亡的诱导被抑制约20%,说明半胱天冬酶依赖的途径参与了(R)-roscovitine对Hela细胞凋亡的诱导;(R)-roscovitine诱导凋亡诱导因子(AIF)从线粒体释放细胞核中,且Ac-DEVD-CHO和Z-VAD-FMK没有抑制AIF的释放,这意味着(R)-roscovitine能够诱导Hela细胞凋亡,其机制是通过caspase与非caspase依赖两种途径.Hela cells used as a model,investigate that the cell apoptosis pathways induced by cyclin dependent kinase inhibitor（R）-roscovitine and further understand that molecular mechanism by which（R）-roscovitinee induced cell apoptosis.The results showed that the proliferation of Hela cells was significantly inhibited by（R）-roscovitine,the cell viability was reduced by 8.48%-33.76%,25.19%-86.41% and 47.90%-88.08%,respectively,after treatment with（R）-roscovitine（17.5-140.0 μmol·L-1） for 12,24 and 48 h.After treatment of Hela cell with（R）-roscovitine for 24 h,the obvious changes in cell morphology occurred by cell shrinkage,round and adherence decrease.Characteristic change of apoptosis with DNA ladder was induced and apoptosis increased with increasing concentration of（R）-roscovitine.Moreover,in the presence of caspase-3 special inhibitor Ac-DEVD-CHO or caspase-pan inhibitor Z-VAD-FMK,the（R）-roscovitine induced apoptosis was reduced by about 20%,suggesting that caspase dependent pathway particaipated in the apoptosis induced by（R）-roscovitine.（R）-roscovitine induced apoptosis inducing factor（AIF）,which could release from mitochondria to nuclei,in the presence or absence of Z-VAD-FMK,demonstrating that（R）-roscovitine could induce apoptosis and its mechanism by which was through both caspase dependent and independent caspase pathways.

关 键 词：(R)-roscovitine 细胞周期蛋白依赖性激酶 细胞凋亡 凋亡诱导因子 半胱天冬酶 

分 类 号：R730.231[医药卫生—肿瘤] R735.2[医药卫生—临床医学]