基于光学表面波技术探究具核梭杆菌介导的自噬与细胞折射率的关系  

Relationship between fusobacterium nucleatum mediated autophagy and cell refractive index based on optical surface wave

作　　者：于丽 王弋嘉[2] 杨勇 刘学焕 万雪花 包翠萍 苗蓓亮 李斯琪 李静 刘筠 YU Li;WANG Yijia;YANG Yong;LIU Xuehuan;WAN Xuehua;BAO Cuiping;MIAO Beiliang;LI Siqi;LI Jing;LIU Jun(School of Integrative Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Translational Medicine Research Institute of Nankai University Peoples Hospital,Tianjin 300121,China;Research Center for Frontier Fundamental Studies,Zhejiang Laboratory,Hangzhou 311100,Zhejiang,China;Teda Institute of Biological Sciences and Biotechnology,Nankai University,Tianjin 300457,China;Department of Nephrology and Endocrinology,Wangjing Hospital,Beijing 100102,China;Department of Radiology,Tianjin Fourth Center Hospital,Tianjin 300142,China)

机构地区：[1]天津中医药大学中西医结合学院,天津301617 [2]南开大学附属人民医院转化医学院,天津300121 [3]之江实验室前沿基础研究中心,浙江杭州311100 [4]南开大学泰达生物技术研究院,天津300457 [5]中国中医科学院望京医院肾病内分泌科,北京100102 [6]天津市第四中心医院影像科,天津300142

出　　处：《山东大学学报（医学版）》2024年第10期87-97,共11页Journal of Shandong University:Health Sciences

基　　金：天津市自然科学基金(21JCYBJC00120);国家自然科学基金(12174203)。

摘　　要：目的拟利用光学表面波传感技术,探索具核梭杆菌(fusobacterium nucleatum,Fn)介导的自噬水平与细胞折射率变化的关系,初步探究光学表面波技术对癌细胞自噬水平检测的潜力。方法运用CCK-8技术检测Fn对结直肠癌细胞增殖活力的影响,蛋白免疫印迹法及自噬双标荧光法检测Fn对自噬水平的影响,同时采用光学表面波技术检测不同Fn感染状态下细胞折射率的变化,观察其与自噬水平的相关性。结果当感染复数为100,感染时间为4 h时,自噬起始蛋白苄氯素1(Beclin-1)、微管相关蛋白1A/1B-轻链3(microtubule-associated protein 1A/1B light chain 3,LC3)蛋白表达水平升高(均P<0.05),螯合体1(sequestome,P62/SQSTM1)蛋白表达水平降低(P<0.01),自噬水平升高;同时光学检测结果显示,HCT116、HT29细胞的折射率亦随自噬水平的增高而升高(均P<0.01)。进一步研究表明,细胞折射率的变化主要在于细胞内自噬小体水平的变化,且不受细胞增殖及胞外物质等因素影响。结论Fn介导的自噬水平与细胞折射率具有相关性,可实时反映Fn介导的自噬水平的变化,表明光学表面波技术可作为一种快速、直观且不影响细胞活性的检测自噬水平的技术。Objective To explore the relationship between autophagy levels mediated by Fusobacterium nucleatum(Fn)and changes in cellular refractive index using optical surface wave sensing technology and to preliminarily investigates the potential of optical surface wave technology for detecting autophagy levels in cancer cells.Methods CCK-8 was used to assess the impact of Fn on the proliferation activity of colorectal cancer cells.The effects of Fn on autophagy levels were examined using both Western blotting and dual-labeling fluorescence microscopy techniques.Additionally,optical surface wave technology was employed to monitor changes in cellular refractive index under different Fn infection states,and to observe their correlation with autophagy levels.Results When multiplicity of infection was 100 and the infection time was 4 hours,the protein expression levels of Beclin-1 and LC3 were increased(P<0.05),while the expression level of P62 was decreased(P<0.01),indicating elevated autophagy levels.Concurrently,optical surface wave showed that the refractive indices of HCT116 and HT29 cells also increased with the elevation of autophagy levels(P<0.01).Further investigation revealed that changes in cellular refractive index were primarily due to variations in autophagosome levels within the cells,and were not influenced by factors such as cell proliferation or extracellular substances.Conclusion The autophagy levels mediated by Fn correlate with cellular refractive index,indicating that optical surface wave sensing technology can dynamically reflect changes in Fn-mediated autophagy levels.This suggests that optical surface wave technology could serve as a rapid,intuitive,and non-invasive method to assess autophagy levels without affecting cellular viability.

关 键 词：具核梭杆菌 结直肠癌 自噬 光学表面波技术 细胞折射率 

分 类 号：Q28[生物学—细胞生物学] R735.3[医药卫生—肿瘤]