Survivin-2B在肿瘤细胞凋亡过程中的作用研究

Study on the role of survivin-2B in apoptosis of tumor cell

目的探讨肿瘤细胞凋亡过程中survivin-2B的作用机制。方法通过流式细胞仪分析survivin-2B对肿瘤细胞周期的影响;利用激光共聚焦显微镜观察通过青色荧光蛋白(CFP)标记的survivin-2B和罗丹明123标记的线粒体在细胞内的定位及在细胞凋亡过程中的变化;采用荧光共振能量转移(fluorescence resonance energy transfer,FRET)方法分析survivin-2B和survivin-ΔEx3在细胞内的相互作用。结果流式细胞仪分析结果表明,Survivin-2B能够明显抑制肿瘤细胞生长,促使细胞凋亡增加;通过激光共聚焦显微镜观察到凋亡早期survivin-2B聚集在细胞发生固缩的部位,且在肿瘤凋亡过程中不存在明显的线粒体定位及与线粒体的相关性;FRET分析显示survivin-2B与survivin-ΔEx3在细胞质中存在很弱的相互作用。结论Survivin-2B能够明显促进肿瘤细胞凋亡,且survivin-2B通过与survivin-ΔEx3直接结合以阻断其抗凋亡效应的可能性较小。

Objective To explore the role of survivin-2B in the process of tumor cell apoptosis.Methods Flow cytometry （FCM） was used to analyze the cycle of HeLa cells transfected with survivin-2B. Survivin-2B were labeled with enhanced cyan fluorescent protein （ECFP）, and the mitochondria were labeled with Rhodamine 123. The cellular location and change during apoptosis of them were subsequently observed with laser confocal scanning microscope. The interaction of survivin-2B and survivin-△Ex3 was analyzed by fluorescence resonance energy transfer （FRET）.Results FCM analysis indicated that survivin-2B could depress the growth and promote the apoptosis of HeLa cells. CLSM observation showed that survivin-2B gathered in the original cellular pyknosis sites during initial phase of apoptosis, and didn＇t display mitochondrial localization and mitochondrial correlation. It was found that there was very weak interaction between survivin-2B and survivin-△Ex3 by FRET analysis.Conclusion Survivin-2B could promote the apoptosis of HeLa cells. It is unlikely for survivin-2B to block the anti-apoptotic effect of survivin-△Ex3 by directly binding to it.