脂滴特异性荧光探针的构建及可视化肝肿瘤细胞

doi:10.6023/cjoc202309008

摘要/Abstract

脂滴(LDs)参与了众多活细胞的代谢过程, 其异常与多种疾病如脂肪肝、糖尿病和癌症等高度相关, 但是准确阐明各种疾病与脂滴之间的相互关系仍然是一个巨大的挑战, 尤其是借助异常分布的LDs实现对肝肿瘤的可视化研究报道较少. 因此, 本论文以三苯胺为供电子基团, 多氰基杂环结构为电子受体, 通过简单的两步反应合成了一种具有LDs特异性的荧光探针TCN-Tph. 制备的探针分子TCN-Tph与脂滴的共定位成像实验表明, 探针TCN-Tph对肝肿瘤细胞具有良好的LDs靶向能力. 采用不同细胞株对探针分子TCN-Tph的响应实验结果表明, 肝肿瘤细胞对其有很好的响应和摄取能力. 不同状态的肝组织切片成像结果进一步表明, 制备的探针分子TCN-Tph对正常肝组织和肝肿瘤组织具有良好的区分功能, 为实现临床上肝肿瘤的早期预诊提供了实验基础.

关键词: 脂滴, 荧光探针, 肝肿瘤细胞, 可视化

Lipid droplets (LDs) are involved in the metabolic processes of many living cells, and their anomalies are highly associated with a variety of diseases, such as fatty liver, diabetes and cancers. But it is still a great challenge to elucidate the intrinsic relationship between various diseases and LDs. In particular, there has been only a few studies on the visualization of abnormal LDs in liver tumor cells. Therefore, in this paper, a lipid droplets targeted fluorescent probe was easily prepared through a two-step procedure by using triphenylamine as the electron donor and polycyanide heterocyclic compound as the electron acceptor. The fluorescent probe TCN-Tph possesses good ability to target LDs in living cells through their co- localization imaging experiments. In comparison with other carcinoma cells, the liver tumor cells have good response and uptake of TCN-Tph. The slices images of liver tissues in different states further show that the probe TCN-Tph can also distinguish liver tissues and normal liver tissues effectively, which provide an experimental basis for the early diagnosis of liver tumor cells in clinic.

Key words: lipid droplets, fluorescent probe, liver tumor cell, visualization

引用此文

徐冬青, 童海姗, 沈杰, 邱万伟, 钱立生. 脂滴特异性荧光探针的构建及可视化肝肿瘤细胞[J]. 有机化学, 2024, 44(4): 1240-1246.

Dongqing Xu, Haishan Tong, Jie Shen, Wanwei Qiu, Lisheng Qian. Construction of a Lipid Droplets Targeted Fluorescent Probe for Visualization of Liver Tumor Cells[J]. Chinese Journal of Organic Chemistry, 2024, 44(4): 1240-1246.

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图/表 8

图式1. 荧光探针TCN-Tph的合成路线.

Scheme 1. Synthesis route of fluorescent probe TCN-Tph

图1. (a) 5.0 μmol/L的探针TCN-Tph在PBS溶液中的归一化紫外和荧光光谱图探针和(b) 5.0 μmol/L的探针TCN-Tph在不同介质溶液中的荧光光谱图

Figure 1. (a) Normalization intensity of UV-Vis absorption and fluorescence spectra for TCN-Tph in PBS solution with the concentration of TCN-Tph at 5.0 μmol/L, and (b) fluorescence spectra of TCN-Tph in different mediums with the concentration of TCN-Tph at 5.0 μmol/L λex=440 nm and λem=560~850 nm. OA: oleic acid, CH: cholesterol, CE: cholesteryl ester, MAG: monoglycerol, DAG: diacylglycerol, TAG: Triacylglycerol, Hcys: homocysteine, GSH: glutathione, Cys: cystein.

图2. 5.0 μmol/L探针TCN-Tph在不同溶剂中的荧光光谱图

Figure 2. Fluorescence spectra of probe TCN-Tph at 5.0 μmol/ L in different solvents λex=440 nm and λem= 650~760 nm.

图3. 荧光探针TCN-Tph对不同细胞株存活率的影响

Figure 3. Effect of fluorescent probe TCN-Tph on livability of living cells (a) HeLa, MAD-MB-231 and MCF-7; (b) Hep G2

图4. 5.0 μmol/L探针TCN-Tph与肝肿瘤细胞中的脂滴共定位荧光成像图

Figure 4. Co-localization fluorescence images of 5.0 μmol/L TCN-Tph in liver tumor cell with LDs TCN-Tph: λex=405 nm and λem=700~800 nm. HCS Lipid TOXTM: λex=637 nm and λem=640~670 nm.

图5. 5.0 μmol/L的探针TCN-Tph在不同细胞中与脂滴的共定位荧光成像图

Figure 5. Co-localization fluorescence images of TCN-Tph at 5.0 μmol/L in different cells with LDs

图6. 探针TCN-Tph在不同细胞中所对应的荧光强度值

Figure 6. Fluorescence intensities of TCN-Tph in different cells corresponding to the fluorescence images TCN-Tph: λex=405 nm and λem=700~800 nm

图7. 3.0 μmol/L探针TCN-Tph在正常肝组织、脂肪肝组织和肝肿瘤组织中的荧光成像图像

Figure 7. Fluorescence images of TCN-Tph at 3.0 μmol/L in normal liver, fat liver and liver tumor tissue TCN-Tph: λex=405 nm and λem=700~800 nm; HCS Lipid TOXTM: λex=637 nm and λem=640~670 nm; Hoechst 33342: λex=405 nm and λem=445~475 nm.

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