[Skp2和p27(kip1)在Jurkat细胞增殖过程中的表达变化及意义]
[Expression variation and significance of Skp2 and p27(kip1) during the proliferation of Jurkat cells].
作者信息
Lu Jian-Xin, Wang Yu-Chan, Shen Ai-Guo, Zhao Yue-Ming, Sun Cheng-Long, Zhang Dong-Mei, Cheng Chun
机构信息
Department of Microbiology and Immunology, Nantong University, Nantong 226001, China.
出版信息
Zhonghua Zhong Liu Za Zhi. 2008 May;30(5):330-4.
OBJECTIVE
To investigate the expression variation and significance of Skp2 and p27(kip1) during the proliferation of lymphoma cell line Jurkat cells.
METHODS
The binding of p27(kip1) 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 p27(kip1) and Skp2 were detected by subcellular fractionation, Western blot and double immunofluorescence labelling.
RESULTS
The results of immunoprecipitation suggested that p27(kip1) and Skp2 could bind each other in Jurkat cells. During the proliferation of Jurkat cells, the protein expression of p27(kip1) decreased and intranuclear p27(kip1) 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 p27(kip1) degradation via the ubiquitin-proteasome pathway, then intranuclear p27(kip1) decreases significantly, leading to an increased cell cycling activity.
目的
研究Skp2和p27(kip1)在淋巴瘤细胞系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p27(kip1)与Skp2的结合情况。对Jurkat细胞进行血清饥饿及释放同步化处理。采用亚细胞分级分离、蛋白质免疫印迹法（Western blot）及双免疫荧光标记法检测p27(kip1)和Skp2的表达变化及亚细胞定位。
结果
免疫沉淀结果提示，p27(kip1)与Skp2在Jurkat细胞中可相互结合。在Jurkat细胞增殖过程中，p27(kip1)蛋白表达下降，核内p27(kip1)显著减少，而Skp2蛋白增加，胞质Skp2显著增加。
结论
在Jurkat细胞增殖过程中，Skp2胞质合成增加可能通过泛素-蛋白酶体途径加速p27(kip1)降解，进而核内p27(kip1)显著减少，导致细胞周期活性增强。
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