THE EFFECTS OF MASS SPECTROMETRY ANALYSIS OF LIPIDOMICS IN CIRCULATING TUMOR CELLS
Victoria Pinilla Escobar1, Richard Cote2.
1University of Miami, Coral Springs, FL, 2University of Miami Miller School of Medicine, Miami, FL.
Circulating tumor cells (CTCs) are cells that have shed into the vasculature from a primary tumor and circulate through the bloodstream, and they are thought to cause metastatic disease. Even though CTCs are present in low numbers in the blood, their characterization, specifically finding a unique signature of phytosterols (PS) and cholesterol lipid profiles as compared with the lipid profiles of primary tumor and metastasis, will potentially provide information to understand if lipids in CTCs influence the metastatic process. In a proof-of-concept study, we began creating a protocol to extract the lipids from the primary tumor, CTCs, and metastasis. Through the use of density gradient centrifugation of the blood, the CTCs and peripheral blood mononucleated cells (PBMCs) were isolated from cancer and healthy patients’ blood. The density gradient centrifugation gives a combination of the CTCs and PBMCs sampled. Therefore, a modified Blight and Dyer method for lipid extraction is currently being tested to see if only the lipids from CTCs can be extracted and analyzed using a triple stage quadruple mass spectroscopy (TSQ-MS) machine. Results presented in this work will serve as a starting point to define possible PS and cholesterol biomarkers that can reveal metabolic pathways with potential for novel drug cancer therapy.