Subcellular localization of the chemotherapeutic agent doxorubicin in renal epithelial cells and in tumor cells using correlative light and electron microscopy

Issue title: Selected papers of the 38th Conference of the German Society for Clinical Microcirculation and Hemorheology, 21-23 November 2019, Braunschweig, Germany

Guest editors: P. Wiggermann, A. Krüger-Genge and F. Jung

Article type: Research Article

Authors: Asam, Claudia^a | Buerger, Korbinian^a | Felthaus, Oliver^c | Brébant, Vanessa^c | Rachel, Reinhard^b | Prantl, Lukas^c | Witzgall, Ralph^a | Haerteis, Silke^{a; *} | Aung, Thiha^c

Affiliations: [a] Institute for Molecular and Cellular Anatomy, University of Regensburg, Germany | [b] Centre for Electron Microscopy, Faculty of Biology and Preclinical Medicine, University of Regensburg, Germany | [c] Centre of Plastic, Aesthetic, Hand and Reconstructive Surgery, University of Regensburg, Regensburg, Germany

Correspondence: [*] Corresponding author: Silke Haerteis, Institute for Molecular and Cellular Anatomy, University of Regensburg, Universitätsstr. 31, 93053 Regensburg, Germany. E-mail: [email protected].

Abstract: BACKGROUND:Doxorubicin is a cytostatic drug from the group of anthracycline antibiotics that is widely used as a chemotherapeutic agent. Side effects of the active substance include cardiotoxicity and nephrotoxicity. Doxorubicin-treated renal epithelial cells and (sarcoma) tumors are examined by correlative light and electron microscopy (CLEM) to investigate the subcellular localization of doxorubicin. METHODS:The kidney epithelial cell line MDCK II (Madin-Darby Canine Kidney) grown on culture dishes were treated with doxorubicin. Subsequently, the cells are analyzed by means of fluorescence and transmission electron microscopy (TEM). In vivo, alveolar rhabdomyosarcoma (RH 30) tumor cells are transferred to the chorioallantoic membrane (CAM) of the chicken embryo. Doxorubicin is injected into a vein of the chicken embryo. After 24 hours, the tumor is removed and examined using CLEM. RESULTS:The kidney epithelial cells and the doxorubicin-injected tumors show a clear staining of the cell nucleus, which correlates with electron-dense regions (heterochromatin). High-resolution TEM shows that doxorubicin treatment leads to an enormous stress situation with an increased formation of membrane blebbings. CONCLUSIONS:CLEM is a promising new method to visualize the pattern of fluorescing drugs (e.g. doxorubicin) in renal epithelial cells and tumors, and to localize the drug in its subcellular context combined with high resolution.

Keywords: Correlative light and electron microscopy (CLEM), chorioallantoic membrane (CAM), doxorubicin, chemotherapy, tumor, sarcoma, Epon

DOI: 10.3233/CH-199212

Journal: Clinical Hemorheology and Microcirculation, vol. 73, no. 1, pp. 157-167, 2019