电化学发光成像分析法监测细胞表面粘蛋白及细胞形貌  

Electrogenerated chemiluminescence imaging method for monitoring MUC1 on cell surface and cellular morphology

作　　者：王晓飞 张成孝[1] 漆红兰[1] WANG Xiaofei;ZHANG Chengxiao;QI Honglan(Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province,School of Chemistry and Chemical Engineering,Shaanxi Normal University,Xi’an 710119,China)

机构地区：[1]陕西省生命分析化学重点实验室,陕西师范大学化学化工学院,陕西西安710119

出　　处：《延安大学学报（自然科学版）》2022年第3期18-23,共6页Journal of Yan'an University：Natural Science Edition

基　　金：国家自然科学基金项目(22074087,21974081);中央高校基础研究经费项目(GK202202002)。

摘　　要：在分子水平上理解细胞功能十分关键。文章以MCF-7细胞为目标细胞,粘蛋白(MUC1)为目标蛋白质,MUC1的特异性适配体为分子识别物质,建立了一种电化学发光成像分析新方法,监测外界刺激下特定细胞、细胞表面粘蛋白含量及细胞形貌的动态变化。结合生物条形码信号放大和Ru(phen)_(3)^(2+)嵌入双链DNA信号转换技术,电化学发光成像灰度值与MUC1含量在8~2000 pg/mL范围内呈良好的线性关系,检出限为2.1 pg/mL;与MCF-7细胞个数在50~1.0×10^(4)cells/mL范围内呈良好的线性关系,检出限为11 cells/mL。进一步应用该方法实时监测不同药物刺激下MCF-7细胞表面MUC1含量和细胞形貌的动态变化。结果发现芹黄素刺激下细胞表面MUC1含量降低,细胞皱缩,尺寸变小;过氧化氢刺激下细胞表面MUC1含量不变,细胞先膨胀,随后缩小。该方法为从细胞水平上灵敏分析蛋白质含量和细胞形貌提供了新思路,有助于理解不同生理过程中细胞形貌和分子机制的变化。Understanding cellular functions on a molecular level is crucial.Here,an electrogenerated chemiluminescence(ECL)imaging method was developed for visualizing and monitoring MUC1 on MCF-7 cell and cellular morphology under external stimulations.As a proof of concept,MCF-7 cells were used as target cells,mucin(MUC1)was used as target protein and anti-MUC1 specific aptamer(rcDNA)was used as molecular recognition element.rcDNA was modified onto the surface of gold electrode and then combined with target MUC1 or MCF-7 cells and hpDNA/AuNP/rcDNA.After Ru(phen)_(3)^(2+)was intercalated into hpDNA,strong ECL signals were generated in the presence of coreactants.The ECL intensity increased with the increase of MUC1 concentration in the range of 8~2000pg/mL with a detection limit of 2.1 pg/mL.The ECL intensity increased with the increase of MCF-7 cells concentration in the range of 50~1×10^(4)cells/mL with a detection limit of 11 cells/mL.In addition,the developed ECL imaging method was used to monitor MUC1 on MCF-7 cells surface and cellular morphology under external drug stimulation,including apigenin and H_(2)O_(2).The decline of MUC1 expression on MCF-7 cells surface and cell shrinkage were observed under apigenin treatment while unconspicuous of MUC1 expression and obvious cell morphological changes were obtained under H_(2)O_(2) treatment.The proposed method provides a new pathway on a molecular level to sensitively analyze protein content and cellular morphology,conducive to understanding cellular morphology and molecular mechanism in different physiological processes.

关 键 词：电化学发光成像 生物传感 细胞 粘蛋白 药物刺激 

分 类 号：O647[理学—物理化学]