Abstract 1195: Ethanolamine kinase-1 and phosphoethanolamine are potential diagnostic markers and therapeutic targets in breast cancer

Increased phosphoethanolamine (PE) has been observed in tumors almost as consistently as increased phosphocholine (PC), but the role of PE in cancer is relatively unexplored. Our ongoing studies have shown that choline kinase (ChK)-α, the enzyme that forms PC from choline, has a dual choline/ethanolamine kinase activity but ChK-β has no significant role in maintaining PE levels in vivo. We observed that ethanolamine kinase (EtnK)-1siRNA significantly reduces PE levels in triple negative MDA-MB-231 human breast cancer cells. We further investigated the role of ethanolamine kinase-2 (EtnK-2) in contributing to the increased PE observed in cancers, and the effect of various small interfering RNA (siRNA) combinations downregulating ChK and EtnK on cell viability, as potential therapeutic strategies in these triple negative breast cancer cells.Cells were cultured in RPMI-1640 medium supplemented with 20μM choline and 50μM ethanolamine. While 50nM siRNA was used in all individual siRNA treatment, for combination siRNA treatment 50nM of each specific siRNA was used. Silencing of 80-90% message was confirmed using qRT-PCR for the siRNAs used. Approximately 40 million cells were harvested 48h post transfection and cell extracts were prepared using a dual-phase extraction method based on methanol/chloroform/water (1/1/1; v/v/v). 31P MR spectra were acquired on water soluble fraction and metabolites were quantitated. For assaying cell viability, 4000 cells per well were plated in a ...
Source: Cancer Research - Category: Cancer & Oncology Authors: Tags: Molecular and Cellular Biology Source Type: research