中国肿瘤生物治疗杂志2025，Vol.32Issue(3)：239-246,8.DOI:10.3872/j.issn.1007-385x.2025.03.002
PD-1高表达细菌质膜囊泡的构建及对小鼠肺癌移植瘤组织的靶向性评估
Construction of PD-1 overexpressing bacterial cytoplasmic membrane vesicles and evaluation of its targeting efficacy of mouse lung cancer xenograft tissue
摘要
Abstract
Objective:To construct bacterial cytoplasmic membrane nanovesicles(BMV)with overexpressing programmed death 1(PD-1),denoted as BMV-PD-1 and evaluate the targeting efficacy of BMV-PD-1 towards transplanted lung tumor tissues in mice.Methods:The fusion plasmid ClyA-PD-1-EGFP fused by PD-1 and Cytolysin A(ClyA)was transferred into Escherichia coli BL21-Codonplus through plasmid transformation.Laser confocal microscopy,SDS-PAGE,and WB were used to detect the expression of the fusion protein ClyA-PD-1-EGFP.Bacterial membranes were extracted and processed with an extruder to generate BMV-PD-1.TEM and NTA were utilized to assess the morphology,size distribution,and zeta potential of BMV-PD-1,while WB was used to verify the presence of PD-1 protein.Laser confocal imaging was conducted to monitor the uptake of BMV-PD-1 by Lewis lung cancer cells.A C57BL/6J mouse subcutaneous transplant tumor model of LLC lung cancer cells was constructed,and the tumor targeting of BMV-PD-1 was evaluated by small animal imaging system.Results:Laser confocal microscopy images demonstrated that the plasmid ClyA-PD-1-EGFP was transferred into BL21-Codonplus and successfully expressed into protein.SDS-PAGE results suggested that ClyA-PD-1-EGFP was overexpressed in BL21-Codonplus.WB analysis indicated that PD-1 was expressed in bacteria and highly expressed in BMV-PD-1(P<0.001).NTA and TEM analyses revealed that BMV-PD-1 were spherical vesicles with a diameter of(145±14)nm and a negative surface charge.Laser confocal imaging showed that the high expression of PD-1 significantly increased the uptake of BMV-PD-1 by lung cancer cells(P<0.01).In vivo imaging of small animals further confirmed that the high expression of PD-1 can effectively improve cancer targeting of BMV-PD-1(P<0.01).Conclusion:In this study,bacterial plasma membrane nanovesicles BMV-PD-1 with high PD-1 expression are successfully constructed,and it is found that PD-1 overexpression markedly improve the mouse lung cancer xenograft tissue targeting specificity of BMV-PD-1,laying the groundwork for further development of BMV-PD-1 as a carrier for targeted drug delivery systems in tumors.关键词
PD-1/细菌囊泡/细胞摄取/肺癌靶向Key words
PD-1/bacterial vesicle/cell uptake/lung cancer targeting分类
医药卫生引用本文复制引用
徐秀杰,兰天舒,张静云,范峻晨,蒋灵馨,张娜,郑猛超,龙雨飞,高贵花,颜涛玲..PD-1高表达细菌质膜囊泡的构建及对小鼠肺癌移植瘤组织的靶向性评估[J].中国肿瘤生物治疗杂志,2025,32(3):239-246,8.基金项目
厦门医学院科技计划项目学校-医院联合资助项目(No.K2023-04) （No.K2023-04）
厦门医学院大学生创新创业训练计划资助项目(No.202412631001) （No.202412631001）
