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目的 研究SNAP 2 3蛋白在NK细胞系NK92细胞胞吐效应中的作用 ,探讨NK细胞胞吐的分子机制。方法 利用已糖氨酶 (β hexosaminidase)释放实验检测NK细胞的胞吐作用。通过RT PCR和Westernblot显示NK92细胞中SNAP 2 3amRNA和蛋白的表达 ,利用激光共聚焦显微镜扫描术确定靶细胞触发前后SNAP 2 3α在NK92细胞中的定位。结果 NK细胞的胞吐作用在靶细胞刺激后 2h明显 ,4～ 6h达高峰 ;NK92细胞组成性表达SNAP 2 3amRNA和蛋白 ,靶细胞刺激前后蛋白表达量虽无明显差异 ,但其定位发生了显著改变 :激光共聚焦显微镜扫描结果显示在静息NK92中SNAP 2 3a主要表达在胞浆内的颗粒上 ,经靶细胞刺激后逐渐向细胞膜发生转位 ,2h明显转位 ,4～ 6h几乎完全转位。结论 NK细胞的胞吐效应与SNAP 2 3蛋白转位相关。
Objective To study the role of SNAP 2 3 in the exocytosis of NK cell line NK92 and explore the molecular mechanism of NK cell exocytosis. Methods The exocytosis of NK cells was detected by β hexosaminidase release assay. The expression of SNAP 2 3 mRNA and protein in NK92 cells was detected by RT PCR and Western blotting. The localization of SNAP 2 3α in NK92 cells was detected by confocal laser scanning microscopy. Results The exocytosis of NK cells was obvious at 2h after stimulation of target cells and peaked at 4 ~ 6h. The expression of SNAP 2 3amRNA and protein was constitutively expressed in NK92 cells, but the expression of SNAP 2 3amRNA was not significantly different Changes: Laser scanning confocal microscopy showed that SNAP 2 3a mainly expressed in the intracytoplasmic granules in resting NK92, and translocated to the cell membrane gradually after stimulation by target cells, and translocated to the cell membrane at 2h after translocation, almost completely after 4 to 6 hours Bit. Conclusion The exocytosis of NK cells is related to the translocation of SNAP 2 3 protein.