Cx30基因的慢病毒质粒构建及其对高糖环境耳蜗毛细胞活力影响
Journal: Basic Medical Theory Research DOI: 10.12238/bmtr.v8i2.18917
Abstract
目的：构建过表达Cx30基因的慢病毒质粒，研究Cx30对高糖诱导耳蜗毛细胞凋亡、增殖影响。方法：本研究通过慢病毒载体构建了携带Flag标签过表达CX30的复合质粒，利用瞬时转染技术过表达CX30、通过CCK-8实验检测细胞活力，流式细胞术检测细胞线粒体膜电位与细胞凋亡。结果：成功构建CX30真核表达质粒。在HEI-OC1细胞中建立NC组和过表达CX30组，在25mmol/L条件下，NC和CX30组细胞数差异显著（P<0.01），CCK-8实验显示，在25mmol/L浓度时，CX30组OD490nm高于NC组（P<0.05）。流式细胞检测细胞凋亡：在25mmol/L浓度时，NC组晚期凋亡率3.4%；CX30组晚期凋亡率4.72%。激光共聚焦显微镜成像显示CX30与PLVAP空间共定位于细胞质及细胞膜。结论：过表达CX30可能通过调控PLVAP抑制凋亡，减轻高糖诱导耳蜗毛细胞损伤。
Keywords
CX30；慢病毒包装；细胞凋亡；增殖；HEI-OC1
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