title: Enhanced detection of metastatic prostate cancer cells in human plasma with lipid bodies staining.
abstract: BACKGROUND
  Reprogramming of energy metabolism of malignant cancer cells confers competitive advantage in growth environments with limited resources. However, not every process of cancer development is associated with competition for resources. During hematogenous transport, cancer cells are exposed to high levels of oxygen and nutrients. Does energy metabolism of cancer cells change as a function of exposure to the bloodstream? Could such changes be exploited to improve the detection of circulating tumor cells (CTC)? These questions have clinical significance, but have not yet been sufficiently examined.
  
  
  METHODS
  The energy metabolism was examined as a function of incubation in nutrient-rich plasma in prostate metastatic cancer cells LNCaP and non-transformed prostate epithelial cells RWPE1. Uptake kinetics of a fluorescent glucose analog (2-NBD) and lipophilic dyes (DiD & Bodipy) were measured in both cell lines, as well as in peripheral blood mononuclear cells (PBMC).
  
  
  RESULTS
  LNCaP cells exhibited hyper-acetylation of low molecular weight proteins compared to RWPE1 cells. Following plasma incubation, protein lysine acetylation profile was unchanged for LNCaP cells while significantly altered for RWPE1 cells. O-linked glycosylated protein profiles were different between LNCaP and RWPE1 cells and varied in both cell lines with plasma incubation. Maximal respiration or glycolytic capacities was unchanged in LNCaP cells and impaired in RWPE1 cells following plasma incubation. However, the uptake rates of 2-NBD and DiD were insufficient for discrimination of LNCaP, or RWPE1 cells from PBMC. On the other hand, both RWPE1 and LNCaP cells exhibited intracellular lipid bodies following plasma incubation; whereas, PBMC did not. The presence of lipid bodies in LNCaP cells permitted retention of Bodipy dye and allowed discrimination of LNCaP cells from PBMC with flow cytometry.
  
  
  CONCLUSIONS
  Despite clear differences in energy metabolism, metastatic prostate cancer cells could not be efficiently distinguished from non-transformed prostate epithelial cells using fluorescent glucose or lipid uptake kinetics. However, metastatic prostate cancer cells in plasma could be clearly distinguished from blood nucleated cells due to the presence of intracellular lipid bodies. Fluorescent labeling of lipid bodies permitted a simple and sensitive means for high throughput detection of metastatic prostate cancer cells in human plasma.
authors: Ranjana
authors: R
authors: Mitra
authors: Roseman University of Health Sciences, 11 Sunset Way, Henderson, NV 89014, USA. ogoodman@roseman.edu.
authors: Oscar B
authors: OB
authors: Goodman
authors: Thuc T
authors: TT
authors: Le
mesh_terms: D002471:Cell Transformation, Neoplastic / Q000737:chemistry / Q000473:pathology*; D005456:Fluorescent Dyes; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry / Q000473:pathology; D008297:Male; D008563:Membrane Lipids / Q000097:blood* / Q000737:chemistry; D009360:Neoplastic Cells, Circulating / Q000737:chemistry / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000097:blood / Q000737:chemistry / Q000473:pathology*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D008563:Membrane Lipids