Analytical and bioanalytical chemistry, 2010-12, Vol.398 (7-8), p.3051-3061
2010
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- Autor(en) / Beteiligte
- Titel
- Raman chemical mapping reveals site of action of HIV protease inhibitors in HPV16 E6 expressing cervical carcinoma cells
- Ist Teil von
- Analytical and bioanalytical chemistry, 2010-12, Vol.398 (7-8), p.3051-3061
- Ort / Verlag
- Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag
- Erscheinungsjahr
- 2010
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- It has been shown that the HIV protease inhibitors indinavir and lopinavir may have activity against the human papilloma virus (HPV) type 16 inhibiting HPV E6-mediated proteasomal degradation of p53 in cultured cervical carcinoma cells. However, their mode and site of action is unknown. HPV-negative C33A cervical carcinoma cells and the same cells stably transfected with E6 (C33AE6) were exposed to indinavir and lopinavir at concentrations of 1 mM and 30 μM, respectively. The intracellular distribution of metabolites and metabolic changes induced by these treatments were investigated by Raman microspectroscopic imaging combined with the analysis of cell fractionation products by liquid chromatography-mass spectrometry (LC-MS). A uniform cellular distribution of proteins was found in drug-treated cells irrespective of cell type. Indinavir was observed to co-localise with nucleic acid in the nucleus, but only in E6 expressing cells. Principal components analysis (PCA) score maps generated on the full Raman hypercube and the corresponding PCA loadings plots revealed that the majority of metabolic variations influenced by the drug exposure within the cells were associated with changes in nucleic acids. Analysis of cell fractionation products by LC-MS confirmed that the level of indinavir in nuclear extracts was approximately eight-fold greater than in the cytoplasm. These data demonstrate that indinavir undergoes enhanced nuclear accumulation in E6-expressing cells, which suggests that this is the most likely site of action for this compound against HPV.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1618-2642eISSN: 1618-2650DOI: 10.1007/s00216-010-4283-6Titel-ID: cdi_proquest_miscellaneous_812127689
- Format
- –
- Schlagworte
- Analytical Chemistry, Biochemistry, Cell Fractionation, Cell Line, Tumor, Cellular, Characterization and Evaluation of Materials, Chemistry, Chemistry and Materials Science, Chromatographic methods and physical methods associated with chromatography, Chromatography, Liquid - methods, Exact sciences and technology, Female, Food Science, Fractionation, HIV, HIV Protease Inhibitors - pharmacology, HPV, Human immunodeficiency virus, Human papillomavirus 16, Human papillomavirus 16 - isolation & purification, Humans, Hypercubes, Imaging, Indinavir, Indinavir - pharmacology, Laboratory Medicine, LC-MS, Lopinavir, Monitoring/Environmental Analysis, Nucleic acids, Oncogene Proteins, Viral - metabolism, Original Paper, Other chromatographic methods, Papillomavirus Infections - drug therapy, Papillomavirus Infections - metabolism, Papillomavirus Infections - virology, Principal Component Analysis, Protease inhibitors, Pyrimidinones - pharmacology, Raman chemical mapping, Repressor Proteins - metabolism, Spectrometric and optical methods, Spectrometry, Mass, Electrospray Ionization - methods, Spectrum Analysis, Raman - methods, Transfection, Tumor Suppressor Protein p53 - metabolism, Uterine Cervical Neoplasms - drug therapy, Uterine Cervical Neoplasms - metabolism, Uterine Cervical Neoplasms - virology