Label-Free Monitoring of Drug-Induced Cytotoxicity and Its Molecular Fingerprint by Live-Cell Raman and Autofluorescence Imaging

Li, Menglu; Liao, Hao Xiang; Bando, Kazuki; Nawa, Yasunori; Fujita, Satoshi; Fujita, Katsumasa

doi:https://doi.org/10.1021/acs.analchem.2c00293

Label-Free Monitoring of Drug-Induced Cytotoxicity and Its Molecular Fingerprint by Live-Cell Raman and Autofluorescence Imaging

Li, Menglu; Liao, Hao Xiang; Bando, Kazuki et al.

Analytical Chemistry, 2022, 94(28), 10019-10026

Number of Access:139（2026-02-18 23:54 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/103305

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Title	Label-Free Monitoring of Drug-Induced Cytotoxicity and Its Molecular Fingerprint by Live-Cell Raman and Autofluorescence Imaging
Creator	CreatorName Li, Menglu
	CreatorName Liao, Hao Xiang
	NRID 1000040795952 1000040795952 researchmap permalink kazukibando kazukibando Scopus Author ID 11540437400 11540437400 CreatorName Bando, Kazuki
	CreatorName Nawa, Yasunori
	NRID 1000000392655 1000000392655 researchmap permalink read0120077 read0120077 Scopus Author ID 57193804976 57193804976 CreatorName Fujita, Satoshi
	NRID 1000080362664 1000080362664 ORCID 0000-0002-2284-375X 0000-0002-2284-375X researchmap permalink read0089911 read0089911 Scopus Author ID 7404058829 7404058829 CreatorName Fujita, Katsumasa
Description	This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry 94, 10019 (2022), © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.2c00293.
Abstract	Simultaneous observation of drug distribution at the effector site and subsequent cell response are essential in the drug development process. However, few studies have visualized the drug itself and biomolecular interactions in living cells. Here, we used label-free Raman microscopy to investigate drug-induced cytotoxicity and visualize drug uptake and subcellular localization by its specific molecular fingerprint. A redox-sensitive Raman microscope detected the decrease of reduced cytochrome c (cyt c) after Actinomycin D (ActD) treatment in a time-dependent and dose-dependent format. Immunofluorescence staining of cyt c suggested that the release of cyt c was not the major cause. Combining Raman microscopy with conventional biological methods, we reported that the oxidization of cyt c is an early cytotoxicity marker prior to the release of cyt c. Moreover, as the spectral properties of ActD are sensitive to the surrounding environment, subcellular localization of ActD was visualized sensitively by the weak autofluorescence, and the intercalation of ActD into DNA was detected by shifted Raman peaks, allowing for parallel observation of drug uptake and the mechanism of action. In this research, we achieved simultaneous observation of cytotoxicity and cellular drug uptake by Raman microscopy, which could facilitate a precise understanding of pharmacological effects and predict potential drug toxicity in the future.
Publisher	American Chemical Society
Source Title	Analytical Chemistry
Volume (Issue)	94(28)
Page	10019-10026
Date of Issued	2022-07-05
Language	English
Handle URL	https://hdl.handle.net/11094/103305
PISSN	00032700
eISSN	15206882
Relation.isVersionOf	DOI (Publisher Version) https://doi.org/10.1021/acs.analchem.2c00293
Access Rights	open access
oaire:version	Accepted Manuscript
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Analytical Chemistry. 2022, 94(28), p. 10019-10026
DC.title	Label-Free Monitoring of Drug-Induced Cytotoxicity and Its Molecular Fingerprint by Live-Cell Raman and Autofluorescence Imaging
DC.creator	Li, Menglu
	Liao, Hao Xiang
	Bando, Kazuki
	Nawa, Yasunori
	Fujita, Satoshi
	Fujita, Katsumasa
DC.publisher	American Chemical Society
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2022-07-05
DC.identifier	https://doi.org/10.1021/acs.analchem.2c00293
DC.description	This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry 94, 10019 (2022), © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.2c00293.
DCTERMS.abstract	Simultaneous observation of drug distribution at the effector site and subsequent cell response are essential in the drug development process. However, few studies have visualized the drug itself and biomolecular interactions in living cells. Here, we used label-free Raman microscopy to investigate drug-induced cytotoxicity and visualize drug uptake and subcellular localization by its specific molecular fingerprint. A redox-sensitive Raman microscope detected the decrease of reduced cytochrome c (cyt c) after Actinomycin D (ActD) treatment in a time-dependent and dose-dependent format. Immunofluorescence staining of cyt c suggested that the release of cyt c was not the major cause. Combining Raman microscopy with conventional biological methods, we reported that the oxidization of cyt c is an early cytotoxicity marker prior to the release of cyt c. Moreover, as the spectral properties of ActD are sensitive to the surrounding environment, subcellular localization of ActD was visualized sensitively by the weak autofluorescence, and the intercalation of ActD into DNA was detected by shifted Raman peaks, allowing for parallel observation of drug uptake and the mechanism of action. In this research, we achieved simultaneous observation of cytotoxicity and cellular drug uptake by Raman microscopy, which could facilitate a precise understanding of pharmacological effects and predict potential drug toxicity in the future.
DC.format	103305
citation_title	Label-Free Monitoring of Drug-Induced Cytotoxicity and Its Molecular Fingerprint by Live-Cell Raman and Autofluorescence Imaging
citation_author	Li, Menglu
	Liao, Hao Xiang
	Bando, Kazuki
	Nawa, Yasunori
	Fujita, Satoshi
	Fujita, Katsumasa
citation_publisher	American Chemical Society
citation_language	English
citation_date	2022-07-05
citation_journal_title	Analytical Chemistry
citation_volume	94
citation_issue	28
citation_firstpage	10019
citation_lastpage	10026
citation_doi	https://doi.org/10.1021/acs.analchem.2c00293
citation_public_url	https://hdl.handle.net/11094/103305

Label-Free Monitoring of Drug-Induced Cytotoxicity and Its Molecular Fingerprint by Live-Cell Raman and Autofluorescence Imaging

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