激光生物学报2024，Vol.33Issue(2)：151-159,9.DOI:10.3969/j.issn.1007-7146.2024.02.007
肿瘤细胞胞外囊泡DNA特征分析
Analysis of the Characteristics of Extracellular Vesicle DNA of Tumor Cells
摘要
Abstract
Extracellular vesicles(EVs)were isolated from the culture supernatant of lung adenocarcinoma A549 cells.The size of EVs was characterized as approximately 100 nm via DLS and TEM analysis,with the EV-specific marker TSG101 detected in the samples.The extracted EV DNA(evDNA)showed a predominantly～12 kb size distribution as assessed by agarose gel electro-phoresis.Based on sequencing data,specific evDNA was selected for PCR detection.DNase I or PS nuclease treatment of the EVs confirmed that the evDNA existed on the outside of the vesicle in a linear manner,and poly-dA tails were added to evDNA by TdT enzyme treatment,allowing for their capture by oligo-dT beads and presenting further evidence of their linear surface distribution.Additionally,a TetO-DNA/TetR-GFP visualization system was established to confirm the presence of TetO-DNA in the EVs,which could be visualized by TetR-GFP proteins under fluorescent microscopy.Enrichment of EVs carrying the TetO-DNA was confirmed via flow cytometry and PCR experiments using TetR-GFP protein.These findings collectively suggest that evDNA is distributed linearly on the surface of EVs,providing a basis for further research into their biological functions.关键词
肺腺癌细胞/胞外囊泡/evDNA/Tet操纵基因-DNA/Tet阻遏蛋白-绿色荧光蛋白Key words
lung adenocarcinoma cells/extracellular vesicles/evDNA/TetO-DNA/TetR-GFP分类
生物科学引用本文复制引用
陈思宇,章运生,裴超柱,谭拥军..肿瘤细胞胞外囊泡DNA特征分析[J].激光生物学报,2024,33(2):151-159,9.基金项目
国家自然科学基金项目(81773169). （81773169）
