Intracellular dynamics of topoisomerase I inhibitor, CPT-11, by slit-scanning confocal Raman microscopy

Most molecular imaging technologies require exogenous probes and may have some influence on the intracellular dynamics of target molecules. In contrast, Raman scattering light measurement can identify biomolecules in their innate state without application of staining methods. Our aim was to analyze intracellular dynamics of topoisomerase I inhibitor, CPT-11, by using slit-scanning confocal Raman microscopy, which can take Raman images with high temporal and spatial resolution. We could acquire images of the intracellular distribution of CPT-11 and its metabolite SN-38 within several minutes without use of any exogenous tags. Change of subcellular drug localization after treatment could be assessed by Raman imaging. We also showed intracellular conversion from CPT-11 to SN-38 using Raman spectra. The study shows the feasibility of using slit-scanning confocal Raman microscopy for the non-labeling evaluation of the intracellular dynamics of CPT-11 with high temporal and spatial resolution. We conclude that Raman spectromicroscopic imaging is useful for pharmacokinetic studies of anticancer drugs in living cells.

[1]  E. Perrier,et al.  Quantitative Analysis of Vitamin A Degradation by Raman Spectroscopy , 2003, Applied spectroscopy.

[2]  Shufeng Zhou,et al.  Simultaneous determination of irinotecan (CPT-11) and SN-38 in tissue culture media and cancer cells by high performance liquid chromatography: application to cellular metabolism and accumulation studies. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[3]  H. Barr,et al.  Raman spectroscopy: elucidation of biochemical changes in carcinogenesis of oesophagus , 2006, British Journal of Cancer.

[4]  V. Davidson,et al.  Resonance Raman spectroscopy of amicyanin, a blue copper protein from Paracoccus denitrificans. , 1988, The Journal of biological chemistry.

[5]  Yves Pommier,et al.  ABCG2 Mediates Differential Resistance to SN-38 (7-Ethyl-10-hydroxycamptothecin) and Homocamptothecins , 2004, Journal of Pharmacology and Experimental Therapeutics.

[6]  Guojin Zhang,et al.  Imaging the prodrug-to-drug transformation of a 5-fluorouracil derivative in skin by confocal Raman microscopy. , 2007, The Journal of investigative dermatology.

[7]  Hidetoshi Sato,et al.  Development of a direct Raman imaging system for rapid diagnosis of malignant tumor , 2008, SPIE BiOS.

[8]  Paul R. Carey,et al.  Biochemical Applications of Raman and Resonance Raman Spectroscopies , 1982 .

[9]  Satoshi Kawata,et al.  Three-dimensional optical-transfer-function analysis for a laser-scan fluorescence microscope with an extended detector , 1991 .

[10]  K Nomura,et al.  Intracellular distribution of CPT-11 in CPT-11-resistant cells with confocal laser scanning microscopy. , 1992, Japanese journal of clinical oncology.

[11]  I. Jolliffe Principal Component Analysis , 2002 .

[12]  Teizo Kitagawa,et al.  Primary protein response after ligand photodissociation in carbonmonoxy myoglobin , 2007, Proceedings of the National Academy of Sciences.

[13]  L. Doyle,et al.  A multidrug resistance transporter from human MCF-7 breast cancer cells. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Pelton,et al.  Spectroscopic methods for analysis of protein secondary structure. , 2000, Analytical biochemistry.

[15]  Dai Ping,et al.  Imaging of anticancer agent distribution by a slit-scanning Raman microscope , 2008, SPIE BiOS.

[16]  Brian C Wilson,et al.  Diagnostic potential of near-infrared Raman spectroscopy in the colon: differentiating adenomatous from hyperplastic polyps. , 2003, Gastrointestinal endoscopy.

[17]  Mortazavi,et al.  Supporting Online Material Materials and Methods Figs. S1 to S13 Tables S1 to S3 References Label-free Biomedical Imaging with High Sensitivity by Stimulated Raman Scattering Microscopy , 2022 .

[18]  N Ramanujam,et al.  Development of a multivariate statistical algorithm to analyze human cervical tissue fluorescence spectra acquired in vivo , 1996, Lasers in surgery and medicine.

[19]  R. Weissleder,et al.  Molecular imaging in drug discovery and development , 2003, Nature Reviews Drug Discovery.

[20]  Satoshi Kawata,et al.  Raman microscopy for dynamic molecular imaging of living cells. , 2008, Journal of biomedical optics.

[21]  P. Carey CHAPTER 4 – Protein Conformation from Raman and Resonance Raman Spectra , 1982 .

[22]  Katsumasa Fujita,et al.  Label-free biochemical imaging of heart tissue with high-speed spontaneous Raman microscopy. , 2009, Biochemical and biophysical research communications.

[23]  Romuald Pawluczyk,et al.  High-performance dispersive Raman and absorption spectroscopy as tools for drug identification , 2009, BiOS.

[24]  M. Manfait,et al.  Confocal Scanning Microspectrofluorometry Reveals Specific Anthracyline Accumulation in Cytoplasmic Organelles of Multidrug-resistant Cancer Cells , 1998, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[25]  M. Manfait,et al.  Structure-activity relation in camptothecin antitumor drugs: why a detailed molecular characterisation of their lactone and carboxylate forms by Raman and SERS spectroscopies? , 1997, Biochimica et biophysica acta.

[26]  Jian Ling,et al.  Direct Raman imaging techniques for study of the subcellular distribution of a drug. , 2002, Applied optics.