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存活蛋白(survivin)是重要的肿瘤相关抗原基因,在肿瘤的发生发展中起着重要的作用.除了标准的剪接形式外,它至少还编码2种变异剪接产物—存活蛋白-2B和存活蛋白-ΔEx3,这2个变异剪接体所编码的蛋白具有不同的生物学功能.为研究这2个变异剪接体在肿瘤细胞中的相互作用情况,本实验利用增强型青色荧光蛋白(enhanced cyan fluorescent protein,ECFP)和增强型黄色荧光蛋白(enhanced yellowfluorescent protein,EYFP)分别标记存活蛋白-2B和存活蛋白-ΔEx3.首先通过激光共聚焦扫描显微镜观察它们的细胞定位;同时利用荧光共振能量转移(fluorescence resonanceenergytransfer,FRET)技术研究两者在细胞内的相互作用情况.研究结果表明,存活蛋白-2B主要分布在细胞质中,而存活蛋白-ΔEx3则主要分布在细胞核内,少量分布在细胞质中;FRET分析结果显示,两者仅在细胞质中存在着很弱的相互作用,表明两者很可能是通过某种间接的方式发挥功能上的相互调节作用.本研究为进一步探讨存活蛋白变异剪接体的生物学功能及相互作用机制奠定了基础.
Survivin is an important tumor-associated antigen gene that plays an important role in tumorigenesis and development.In addition to the standard splicing forms, it also encodes at least two alternative splicing products, Survivin-2B and Survivin- ΔEx3, the proteins encoded by these two variants have different biological functions.In order to study the interaction between these two variants in tumor cells, we used enhanced cyan fluorescent protein ECFP) and enhanced yellow fluorescent protein (EYFP) were used to label Survivin-2B and Survivin-ΔEx3 respectively.Firstly, their cellular localization was observed by laser confocal scanning microscopy and fluorescence resonance energy transfer (fluorescence resonance energy transfer, FRET) technology to study the interaction between the two in the cell.The results showed that Survivin-2B was mainly distributed in the cytoplasm, while Survivin-ΔEx3 was mainly distributed in the nucleus, a small amount of distribution in the cytoplasm; FRET analysis showed , There is only a weak interaction between the two in the cytoplasm, indicating that both are very good Is able to play a regulatory role on each other’s function in some indirect way. This study further explore the function and survival mechanisms of interaction of biological protein variants spliceosome basis.