A highly selective fluorescent probe for real-time imaging of UDP-glucuronosyltransferase 1A8 in living cells and tissues

doi:10.1007/s11705-021-2064-8

Front. Chem. Sci. Eng.

2022, Vol. 16

Issue (1) : 103-111 https://doi.org/10.1007/s11705-021-2064-8

RESEARCH ARTICLE

A highly selective fluorescent probe for real-time imaging of UDP-glucuronosyltransferase 1A8 in living cells and tissues

Mingyue Zhu^1,², Zhenhao Tian³, Lingling Jin², Xiaokui Huo², Chao Wang², Jingnan Cui⁴, Yan Tian²(

), Xiangge Tian²(

), Lei Feng^1,²(

)

¹. College of Pharmacy, School of Medicine, Key Laboratory of Elemene Class Anti-Cancer Chinese Medicines, Engineering Laboratory of Development and Application of Traditional Chinese Medicines, Hangzhou Normal University, Hangzhou 311121, China
². Dalian Key Laboratory of Metabolic Target Characterization and Traditional Chinese Medicine Intervention, College of Pharmacy, College of Integrative Medicine, Dalian Medical University, Dalian 116044, China
³. School of Life Sciences, Northwestern Polytechnical University, Xi’an 710072, China
⁴. State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China

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Abstract

Uridine diphosphate (UDP)-glucuronosyltransferases (UGTs) are enzymes involved in the biotransformation of important endogenous compounds such as steroids, bile acids, and hormones as well as exogenous substances including drugs, environmental toxicants, and carcinogens. Here, a novel fluorescent probe BDMP was developed based on boron-dipyrromethene (BODIPY) with high sensitivity for the detection of UGT1A8. The glucuronidation of BDMP not only exhibited a red-emission wavelength (λ_ex/λ_em = 500/580 nm), but also displayed an excellent UGT1A8-dependent fluorescence signal with a good linear relationship with UGT1A8 concentration. Based on this perfect biocompatibility and cell permeability, BDMP was successfully used to image endogenous UGT1A8 in human cancer cell lines (LoVo and HCT15) in real time. In addition, BDMP could also be used to visualize UGT1A8 in tumor tissues. These results suggested that BDMP is a promising molecular tool for the investigation of UGT1A8-mediated physiological function in humans.

Keywords UDP-glucuronosyltransferase 1A8 fluorescent probe subtype selectivity fluorescence imaging

Corresponding Author(s): Yan Tian,Xiangge Tian,Lei Feng

Online First Date: 13 July 2021 Issue Date: 27 December 2021

Cite this article:

Mingyue Zhu,Zhenhao Tian,Lingling Jin, et al. A highly selective fluorescent probe for real-time imaging of UDP-glucuronosyltransferase 1A8 in living cells and tissues[J]. Front. Chem. Sci. Eng., 2022, 16(1): 103-111.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-021-2064-8
https://academic.hep.com.cn/fcse/EN/Y2022/V16/I1/103

Fig.1 Scheme 1 The glucuronidation reaction of BDMP mediated by UGT1A8.

Fig.2 (a) Absorption and (b) fluorescence emission spectra of BDMP (10 μmol·L^–1) before and after incubating with UGT1A8; (c) the HPLC analysis of BDMP and the incubation sample with UGT1A8.

Fig.3 (a) The fluorescence emission spectrum of BDMP (10 μmol·L^–1) after incubating with different concentrations of UGT1A8; (b) the linear regression of fluorescence intensity with UGT1A8 concentration.

Fig.4 (a) Fluorescence intensity of BDMP (10 μmol·L^–1) incubating with 13 isoforms of UGTs; (b) the stability of BDMP among metal ions and amino acids; (c) chemical inhibition of different inhibitors on UGT1A8 mediating the glucuronidation reaction; (d) the inhibition curve of Nilotinib toward the BDMP glucuronidation.

Fig.5 The kinetic analysis of BDMP in the presence of UGT1A8.

Fig.6 (a, e, i) The fluorescence of HCT-15 cells; (b, f, j) staining of nuclei by Hoechst 33342; (c, g, k) bright field images of HCT15 cells; (d, h, l) merge of the confocal fluorescence images (The scale bar is 50 μm. BDMP: λ_ex 514 nm; λ_em 550–610 nm).

Fig.7 (a, d, g) Fluorescence microscope imaging of UGT1A8 for the tissue slices of normal LoVo tumor tissues; (b, e, h) nuclear staining with DAPI; (c, f, i) merged confocal fluorescence images (The scale bars are 50 and 25 μm. λ_ex 514 nm; λ_em 550–610 nm).

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