中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2008年
5期
330-334
,共5页
陆建新%王燏婵%沈爱国%赵玥铭%孙承龙%张冬梅%程纯
陸建新%王燏嬋%瀋愛國%趙玥銘%孫承龍%張鼕梅%程純
륙건신%왕율선%침애국%조모명%손승룡%장동매%정순
Jurkat细胞株%p27kip1%Skp2
Jurkat細胞株%p27kip1%Skp2
Jurkat세포주%p27kip1%Skp2
Jurkat cell line%p27kip1%Skp2
目的 探讨S期激酶相关蛋白2(Skp2)与p27kip1在淋巴瘤细胞株Jurkat细胞增殖过程中的表达变化及意义.方法 采用免疫沉淀法检测Jurkat细胞中p27kip1与Skp2的结合情况;采用血清饥饿合并释放法同步化处理Jurkat细胞,再分别用Western blot技术、免疫荧光双标技术及核浆分离法检测p27kip1和Skp2在Jurkat细胞中的表达变化及亚细胞定位.结果 p27kip1与Skp2能够相互结合.在Jurkat细胞增殖过程中,p27kip1蛋白表达减少,其中在细胞核内的减少更明显;Skp2蛋白表达增加,其中在细胞浆中的增加更明显.结论 在Jurkat细胞增殖过程中,Skp2在细胞浆中的合成增加,并可能通过加快p27kip1的泛素化降解使细胞核内p27kip1显著减少,从而推动细胞周期的进程.
目的 探討S期激酶相關蛋白2(Skp2)與p27kip1在淋巴瘤細胞株Jurkat細胞增殖過程中的錶達變化及意義.方法 採用免疫沉澱法檢測Jurkat細胞中p27kip1與Skp2的結閤情況;採用血清饑餓閤併釋放法同步化處理Jurkat細胞,再分彆用Western blot技術、免疫熒光雙標技術及覈漿分離法檢測p27kip1和Skp2在Jurkat細胞中的錶達變化及亞細胞定位.結果 p27kip1與Skp2能夠相互結閤.在Jurkat細胞增殖過程中,p27kip1蛋白錶達減少,其中在細胞覈內的減少更明顯;Skp2蛋白錶達增加,其中在細胞漿中的增加更明顯.結論 在Jurkat細胞增殖過程中,Skp2在細胞漿中的閤成增加,併可能通過加快p27kip1的汎素化降解使細胞覈內p27kip1顯著減少,從而推動細胞週期的進程.
목적 탐토S기격매상관단백2(Skp2)여p27kip1재림파류세포주Jurkat세포증식과정중적표체변화급의의.방법 채용면역침정법검측Jurkat세포중p27kip1여Skp2적결합정황;채용혈청기아합병석방법동보화처리Jurkat세포,재분별용Western blot기술、면역형광쌍표기술급핵장분리법검측p27kip1화Skp2재Jurkat세포중적표체변화급아세포정위.결과 p27kip1여Skp2능구상호결합.재Jurkat세포증식과정중,p27kip1단백표체감소,기중재세포핵내적감소경명현;Skp2단백표체증가,기중재세포장중적증가경명현.결론 재Jurkat세포증식과정중,Skp2재세포장중적합성증가,병가능통과가쾌p27kip1적범소화강해사세포핵내p27kip1현저감소,종이추동세포주기적진정.
Objective To investigate the expression variation and significance of Skp2 and p27kip1 during the proliferation of lymphoma cell line Jurkat cells. Methods The binding of p27kip1 and Skp2 in Jurkat cells were detected by immunoprecipitation. Jurkat cells were treated with serum starvation and release synchronization. The expression variation and subcellular localization of p27kip1 and Skp2 were detected by subcellular fractionation, Western blot and double immunofluorescence labelling. Results The results of immunoprecipitation suggested that p27kip1 and Skp2 could bind each other in Jurkat cells. During the proliferation of Jurkat cells, the protein expression of p27kip1 decreased and intranuclear p27kip1 decreased significantly, while the Skp2 protein increased and cytoplasmic Skp2 increased significantly. Conclusion During the proliferation of Jurkat cells, the increased cytoplasmic synthesis of Skp2 may speed up p27kip1 degradation via the ubiquitin-proteasome pathway, then intranuclear p27kip1 decreases significantly, leading to an increased cell cycling activity.
