枸杞外泌体对非小细胞肺癌A549细胞的增殖抑制和凋亡的调控作用  

Effects of Lycium barbarum exosomes on proliferation inhibition and apoptosis of non-small cell lung cancer A549 cells

作　　者：张丽昀 王泽华 马春燕 ZHANG Liyun;WANG Zehua;MA Chunyan(Key Laboratory of Conservation and Utilization of Characteristic Biological Resources in Western China,Ministry of Education,College of Life Sciences,Ningxia University,Yinchuan 750021,China)

机构地区：[1]宁夏大学生命科学学院西部特色生物资源保护与利用教育部重点实验室,银川750021

出　　处：《生物学杂志》2024年第5期48-53,共6页Journal of Biology

基　　金：国家自然科学基金项目(31660255);宁夏自然科学基金项目(NZ16033)。

摘　　要：为阐明枸杞外泌体对非小细胞肺癌细胞A549的增殖抑制和凋亡的调控作用,通过超速离心法提取枸杞外泌体,通过透射电镜检测外泌体的形态,粒径分析检测外泌体的直径分布。用提取到的枸杞外泌体作用于A549细胞,通过CCK-8法检测枸杞外泌体对A549细胞的增殖影响,EdU检测枸杞外泌体对A549细胞的活性影响,细胞划痕、Transwell检测细胞的迁移效率,细胞克隆法检测细胞的克隆能力。流式细胞术检测细胞的凋亡率,用Western Blot和qRT-PCR法检测细胞凋亡通路的蛋白和基因表达。结果表明,提取的枸杞外泌体在电镜下呈茶托状,大小在40~200 nm,符合外泌体形状特征。与对照组相比,枸杞外泌体组能够浓度依赖性地抑制A549细胞增殖(P<0.01)。不同浓度枸杞外泌体处理组的细胞迁移能力和细胞克隆能力也呈浓度依赖性下降(P<0.01),而细胞凋亡率呈浓度依赖性增多(P<0.01)。枸杞外泌体能够浓度依赖性地上调Cl-Caspase3、p53、Bax等促凋亡因子,下调Bcl-2等抑凋亡因子的蛋白和基因(P<0.01)。结果表明,枸杞外泌体可以抑制A549细胞增殖,促进A549细胞凋亡并且枸杞外泌体可通过线粒体通路诱导细胞凋亡。In order to clarify the effect of Lycium barbarum exosomes on proliferation inhibition and apoptosis of non-small cell lung cancer cell A549,the exosomes were extracted by ultra-fast centrifugation method.The morphology of exosomes was detected by transmission electron microscopy,and the diameter distribution of exosomes was detected by particle size analysis.A549 cells were treated with the extracted L.barbarum exosomes.The effect of L.barbarum exosomes on A549 cells proliferation was detected by CCK-8 method,EdU was used to detect the effect of L.barbarum exosomes on A549 cells,the migration efficiency of cells was detected by cell scratch method and Transwell,the cloning ability of cells was detected by cell cloning method.The apoptosis rate of cells was detected by flow cytometry,Western Blot and qRT-PCR were used to detect the protein and gene expression of apoptosis pathway.The results showed that the extracted exosomes were saucer-like under electron microscope,and the size was 40-200 nm,which was consistent with the shape characteristics of the exosomes.Compared with control group,L.barbarum exosomes group could inhibit A549 cell proliferation in a concentration-dependent way(P<0.01).The cell migration ability and cell cloning ability in different concentrations of L.barbarum berry exosomes decreased in a concentration-dependent manner(P<0.01),while the cell apoptosis rate increased in a concentration-dependent manner(P<0.01).L.barbarum exosomes could up-regulate the pro-apoptotic factors such as Cl-Caspase3,p53 and Bax in a concentration dependent manner,and down-regulate the proteins and genes of anti-apoptotic factors such as Bcl-2(P<0.01).The conclusion was that L.barbarum exosomes could inhibit the proliferation of A549 cells and promote the apoptosis of A549 cells,and L.barbarum exosomes could also induce apoptosis through mitochondrial pathway.

关 键 词：枸杞 外泌体 非小细胞肺癌细胞 增殖抑制 细胞凋亡 

分 类 号：Q25[生物学—细胞生物学] R285[医药卫生—中药学]