ECE2023 Poster Presentations Adrenal and Cardiovascular Endocrinology (72 abstracts)
Non-specific uptake of magnetic iron oxide nanoparticles by Adrenocortical carcinoma and Endothelial cells
Anna Sorushanova 1 , Padraig Donlon 1 , Nathan Mullen 1 , Obdulia Covarrubias-Zambrano 2 , Jose Covarrubias 2 , Sunitha Varghese 2 , Constanza Hantel 3 , Peter Owens 4 , Martin O’Halloran 5 , Punit Prakash 6 , Stefan Bossman 2 & Michael Conall Dennedy 1
1University of Galway, Discipline of Pharmacology & Therapeutics, School of Medicine, Galway, Ireland; 2The University of Kansas Medical Center, Department of Cancer Biology, Kansas, United States; 3University Hospital Zurich, Department of Endocrinology, Zurich, Switzerland; 4University of Galway, Centre for Microscopy & Imaging, Galway, Ireland; 5University of Galway, Translational Medical Device, Galway, Ireland; 6Kansas State University, Department of Electrical and Computer Engineering, Kansas, United States
Adrenocortical carcinoma is a rare malignant tumour that starts in the adrenal gland, accounting for 4% of all adrenal tumours. Functioning ACC can cause symptoms related to the overproduction of hormones and large tumours can press on nearby organs. Primary treatment for ACC tumour is surgery to remove the tumour, and may also include Chemotherapy or Radiation to stop the cancer cells from spreading. Therefore, more affective therapies for delivering therapeutics to the tumour are highly desirable. Magnetic iron oxide nanoparticles (MIONPs) have been gaining traction over the years for treatment of various cancers as they can be produced in various shapes and sizes, with the ability to modify the surface by coating the nanoparticles. Hence, MIONP delivered chemotherapy with associated hyperthermia has become an area of great interest. In this study, MIONPs have been used at different concentrations to evaluate non-specific nanoparticle uptake and rate of uptake by adrenal cortical and endothelial cells, as well as gain understanding of the location of nanoparticles within the cell. MIONPs were provided by The University of Kansas. Adrenal Cortical cell-lines (MUC1, H295R and HAC15) and Endothelial cell-line (HUVEC) were used in this study. Cells were incubated with MIONPs were for 24 h at concentrations ranging from 0.5 to 50 µg/ml. MIONP uptake, rate of uptake and vaibility was assessed by Flow Cytometry. Confocal Microscopy was used to image cellular morphology, and live imaging was carried out to view live uptake. Cellular proliferation was assessed by Ki-67. “Seahorse” technology was used to assess the effect of MIONPs on mitochondrial acttivity. Intracellular location of MIONPs was visualused TEM. The effect on steroidogenesis in the presence of MIONPs was assessed by Mass Spectrometry. Steroidogenic enzyme expression was assessed by RT-PCR. Viability, the location of MIONPs and proliferation post 7 days were assessed by Flow Cytometry, TEM and Ki-67. Following 24 hour incubation, Flow Cytometry showed significant uptake by Adrenocortical and Endothelial cells at 10 µg/ml MIONP concentration. Confocal and TEM images revealed MIONPs reside in the cytoplasm and in the vesicles of the cells. Live Confocal imaging showed movement of HAC15 cells towards MIONPs for phagocytosis. We demonstrated that ACC cells uptake MIONPs in a time and dose dependent manner. MIONP concentration from 0.5 to 10 µg/ml showed no cytotoxic affect, while cytotoxic affect was observed at 50 µg/ml. 10 µg/ml MIONP concentration was deemed optimal for ACC and Endothelial cells.
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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