结直肠癌细胞WS480中硫化氢和生物硫醇的检测与成像研究
Detection and imaging of hydrogen sulfide and biological thiols in colorectal cancer WS480 cells
王元凡 1刘天乐 1缪林熠 2任晔潼 1陈俊凝 3张胜香 3徐洲4
作者信息
- 1. 徐州医科大学第一临床医学院,江苏 徐州 221004
- 2. 徐州医科大学口腔医学院
- 3. 徐州医科大学药学院
- 4. 徐州医科大学药学院;徐州医科大学公共实验研究中心
- 折叠
摘要
目的 旨在设计、合成一种新型荧光探针,用于结直肠癌细胞中生物硫醇和硫化氢的检测与成像,期望为结直肠癌的早期诊断提供新工具和新方法.方法 设计的探针WZL-1 通过 4 步反应从商业易得的廉价原料合成而得,并对其光学性能、细胞毒性以及活细胞荧光成像进行了详细评价.在评价中,考虑了探针的荧光特性、对生物硫醇和硫化氢的敏感性,以及其在活细胞成像中的适用性.结果 在生理条件下探针WZL-1 对生物硫醇半胱氨酸、同型半胱氨酸、谷胱甘肽和硫化氢在 630 nm处的发射强度均显著增强.探针表现出卓越的灵敏度,在仅需 30 s的极短时间内,WZL-1 能够迅速准确地检测目标分子,且荧光强度至少可以维持 50 min.WZL-1 荧光探针的线性范围广泛,其对 GSH的检测线低至 554 nm,探针在 0～30 μmol/L 浓度范围内细胞存活率超过90%.结论 荧光探针WZL-1 能与结直肠癌细胞WS480 内的生物硫醇和硫化氢发生特异性反应,并释放出红色荧光信号,而其他物质分子则不与其反应,其可用于细胞中各种生物硫醇和硫化氢的实时监测与成像.
Abstract
Objective To design and synthesize a novel fluorescent probe for the detection and imaging of biothiols and hydrogen sulfide in colorectal cancer cells,with the expectation of providing new tools and methods for the early diag-nosis of colorectal cancer.Methods The designed probe,WZL-1,was synthesized from commercially available and in-expensive raw materials through a four-step reaction.Its optical properties,cytotoxicity,and fluorescence imaging in liv-ing cells were evaluated in detail.The probe's fluorescence characteristics,sensitivity to biothiols and hydrogen sulfide,and its suitability for live cell imaging were analyzed.Results Under the physiological conditions,the probe WZL-1 ex-hibited significantly enhanced emission intensities at 630 nm for thiols cysteine,homocysteine,glutathione,and hydrogen sulfide.The probe demonstrated excellent sensitivity,and rapidly and accurately detected target molecules within a short period of time of 30 s,with fluorescence intensity maintained for at least 50 min.The WZL-1 fluorescence probe had a wide linear range,with a detection limit as low as 554 nm for GSH.The cell survival rate was more than 90%over the concentration range of 0-30 μmol/L.Conclusions The fluorescent probe WZL-1 can specifically react with bio thiols and hydrogen sulfide in colorectal cancer WS480 cells,emitting red fluorescence signals,while other substances do not react.It can be used for real-time monitoring and imaging of various biothiols and hydrogen sulfide in cells.
关键词
结直肠癌/硫化氢/生物硫醇/荧光探针/检测限/谷胱甘肽Key words
colorectal cancer/hydrogen sulfide/biothiols/fluorescent probe/detection limit引用本文复制引用
基金项目
江苏省自然科学基金面上项目(BK20211059)
江苏省大学生创新创业训练计划重点项目(202310313046Z)
出版年
2024
NETL
NSTL
维普
万方数据