Dataset Information

Tandem fluorescence and Raman (fluoRaman) characterisation of a novel photosensitiser in colorectal cancer cell line SW480.

ABSTRACT: The development of new imaging tools, molecules and modalities is crucial to understanding biological processes and the localised cellular impact of bioactive compounds. A small molecule photosensitiser, DC473, has been designed to be both highly fluorescent and to exhibit a strong Raman signal in the cell-silent region of the Raman spectrum due to a diphenylacetylene structure. DC473 has been utilised to perform a range of novel tandem fluorescence and Raman (fluoRaman) imaging experiments, enabling a thorough examination of the compound's cellular localisation, exemplified in colorectal cancer cells (SW480). This multifunctional fluoRaman imaging modality revealed the presence of the compound in lipid droplets and only a weak signal in the cytosol, by both Raman and fluorescence imaging. In addition, Raman microscopy detected the compound in a cell compartment we labelled as the nucleolus, whereas fluorescence microscopy did not detect the fluoRaman probe due to solvatochromatic effects in a local polar environment. This last finding was only possible with the use of tandem confocal Raman and fluorescence methods. By following the approach detailed herein, incorporation of strong Raman functional groups into fluorophores can enable a plethora of fluoRaman experiments, shedding further light on potential drug compound's cellular behaviour and biological activity.

SUBMITTER: Gala de Pablo J

PROVIDER: S-EPMC6336151 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

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Tandem fluorescence and Raman (fluoRaman) characterisation of a novel photosensitiser in colorectal cancer cell line SW480.

Gala de Pablo Julia J Chisholm David R DR Steffen Andreas A Nelson Amanda K AK Mahler Christoph C Marder Todd B TB Peyman Sally A SA Girkin John M JM Ambler Carrie A CA Whiting Andrew A Evans Stephen D SD

The Analyst 20181201 24

The development of new imaging tools, molecules and modalities is crucial to understanding biological processes and the localised cellular impact of bioactive compounds. A small molecule photosensitiser, DC473, has been designed to be both highly fluorescent and to exhibit a strong Raman signal in the cell-silent region of the Raman spectrum due to a diphenylacetylene structure. DC473 has been utilised to perform a range of novel tandem fluorescence and Raman (fluoRaman) imaging experiments, ena ...[more]

PMID: 30468234

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Project description:Omentin-1, a 34-kDa protein, has been demonstrated to be associated with colorectal cancer (CRC). Epidemiological and clinical studies have indicated that the levels of circulating omentin-1 are significantly increased in patients with CRC, but the cause of the high omentin-1 levels and whether CRC cells express this adipokine have not been determined. In the present study, human colorectal carcinoma and para-carcinoma tissues were collected from 24 patients with CRC. In addition, SW480 and HCT116 colon cancer cells were cultured in vitro. The expression and localization of omentin-1 protein in human CRC and para-carcinoma tissues were determined by immunohistochemistry. The mRNA and protein expression levels of omentin-1 in human CRC and para-carcinoma tissues were detected by reverse transcription-quantitative PCR (RT-qPCR) and western blotting, respectively. In addition, omentin-1 mRNA expression levels in SW480 and HCT116 cell lines were detected by RT-qPCR. Since SW480 cells exhibited higher omentin-1 mRNA levels compared with those of HCT116 cells, SW480 cells were selected for further experiments. The expression of omentin-1 protein in the supernatant and lysate of SW480 cells obtained at 6, 12, 24 and 48 h was determined by ELISA. The immunohistochemistry results demonstrated that the positive expression of omentin-1 protein was mainly located in the cytoplasm of cancer cells in human CRC tissues. The mRNA and protein expression levels of omentin-1 in the CRC tissues were higher compared with those in para-carcinoma tissues. The expression levels of omentin-1 were detected in the cell lysate and supernatant of SW480 cells; the expression level of omentin-1 protein in the cell lysate was higher compared with that in the supernatant. These results indicated that SW480 cells secret and express the adipokine omentin-1 endogenously. Omentin-1 may serve its potential carcinogenetic role in CRC through endocrine, autocrine and paracrine pathways.

Dataset Information

Tandem fluorescence and Raman (fluoRaman) characterisation of a novel photosensitiser in colorectal cancer cell line SW480.

Publications

Tandem fluorescence and Raman (fluoRaman) characterisation of a novel photosensitiser in colorectal cancer cell line SW480.

Similar Datasets

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