HPV Infection Significantly Accelerates Glycogen Metabolism in Cervical Cells with Large Nuclei: Raman Microscopic Study with Subcellular Resolution

• Published in: International journal of molecular sciences Vol. 21; no. 8; p. 2667
• Main Authors: Sitarz, Katarzyna, Czamara, Krzysztof, Bialecka, Joanna, Klimek, Malgorzata, Zawilinska, Barbara, Szostek, Slawa, Kaczor, Agnieszka
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 11.04.2020; MDPI
• Subjects: Carcinoma, Squamous Cell - etiology; Carcinoma, Squamous Cell - metabolism; Carcinoma, Squamous Cell - pathology; Cell Nucleus - metabolism; Cell Nucleus - pathology; Cell size; Cervical cancer; cervical dysplasia; Cervix; Cytoplasm; Depletion; Epithelial Cells - metabolism; Female; Fourier transforms; Genomes; Glycogen; Glycogen - metabolism; Glycogenolysis; Glycogens; Histocytochemistry - methods; Human papillomavirus; Humans; Infections; Intracellular Space - metabolism; Kinases; Medical screening; Menstruation; Metabolism; Metastasis; Microscopy; Mucous Membrane - metabolism; Mucous Membrane - virology; Nonlinear Optical Microscopy; Nuclei (cytology); Papillomavirus Infections - complications; Papillomavirus Infections - virology; Phosphorylation; Protein metabolism; Protein turnover; Proteins; Raman microscopy; Signal transduction; Spectrum analysis; Squamous cell carcinoma; Studies; Uterine Cervical Neoplasms - etiology; Uterine Cervical Neoplasms - metabolism; Uterine Cervical Neoplasms - pathology; Viral infections; Womens health; cervical dysplasia; Raman microscopy; glycogen; glycogenolysis; cervical cancer; human papillomavirus
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21082667

Using Raman microscopy, we investigated epithelial cervical cells collected from 96 women with squamous cell carcinoma (SCC) or belonging to groups I, IIa, IIID-1 and IIID-2 according to Munich III classification (IIID-1 and IIID-2 corresponding to Bethesda LSIL and HSIL groups, respectively). All women were tested for human papillomavirus (HPV) infection using PCR. Subcellular resolution of Raman microscopy enabled to understand phenotypic differences in a heterogeneous population of cervical cells in the following groups: I/HPV , IIa/HPV , IIa/HPV , LSIL/HPV , LSIL/HPV , HSIL/HPV , HSIL/HPV and cancer cells (SCC/HPV ). We showed for the first time that the glycogen content in the cytoplasm decreased with the nucleus size of cervical cells in all studied groups apart from the cancer group. For the subpopulation of large-nucleus cells HPV infection resulted in considerable glycogen depletion compared to HPV negative cells in IIa, LSIL (for both statistical significance, ca. 45%) and HSIL (trend, 37%) groups. We hypothesize that accelerated glycogenolysis in large-nucleus cells may be associated with the increased protein metabolism for HPV positive cells. Our work underlines unique capabilities of Raman microscopy in single cell studies and demonstrate potential of Raman-based methods in HPV diagnostics.