【摘要】 为了研究荞麦胰蛋白酶抑制剂(buckwheat trypsin inhibitor,BTI)对肿瘤细胞凋亡与细胞周期的影响,构建增强型绿色荧光蛋白(EGFP)与BTI融合蛋白真核表达质粒.将BTI基因成功克隆至pEGFP-N1中转染食管癌EC9706细胞后,激光共聚焦显微镜镜检显示,BTI-EGFP获得良好表达.表达的融合蛋白大部分分布于细胞核,在细胞质中有少量分布.Western印迹检测可见约27kD和36 kD的特异性条带.流式细胞术分析结果显示,BTI能够诱导EC9706细胞发生凋亡,并使细胞停滞于G0/G1期.更多还原

【Abstract】 To investigate the effect of buckwheat trypsin inhibitor(BTI) on the cell apoptosis and cell cycle in the tumor cells,we constructed the eukaryotic expression plasmid harboring enhanced green fluorescent protein(EGFP) and BTI fusion protein using a pEGFP-N1 vector.The recombinant was transfected into EC9706 cells afterwards and BTI gene expression was observed well by laser scanning confocal microscopy.The fusion protein mainly distributed in the cellular nucleus,and relative small amount in the cytoplast.Western blot analysis showed approximately 27 kD and 36 kD bands.The results suggested that BTI-EGFP induced EC9706 cell apoptosis and the cell cycle was arrested at G0 /G1 phase by flow cytometry.更多还原