Lipidomic approach to identify patterns in phospholipid profiles and define class differences in mammary epithelial and breast cancer cells

Abstract

Breast cancer is the leading cause of cancer-related deaths in women. Altered cellular functions of cancer cells lead to uncontrolled cellular growth and morphological changes. Cellular biomembranes are intimately involved in the regulation of cell signaling; however, they remain largely understudied. Phospholipids (PLs) are the main constituents of biological membranes and play important functional, structural and metabolic roles. The aim of this study was to establish if patterns in the PL profiles of mammary epithelial cells and breast cancer cells differ in relation to degree of differentiation and metastatic potential. For this purpose, PLs were analyzed using a lipidomic approach. In brief, PLs were extracted using Bligh and Dyer method, followed by a separation of PL classes by thin layer chromatography, and subsequent analysis by mass spectrometry (MS). Differences and similarities were found in the relative levels of PL content between mammary epithelial and breast cancer cells and between breast cancer cells with different levels of aggressiveness. When compared to the total PL content, phosphatidylcholine levels were reduced and lysophosphatydilcholines increased in the more aggressive cancer cells; while phosphatidylserine levels remained unchanged. MS analysis showed alterations in the classes of phosphatidylcholine, lysophosphatidylcholine, sphingomyelin, and phosphatidylinositides. In particular, the phosphatidylinositides, which are signaling molecules that affect proliferation, survival, and migration, showed dramatic alterations in their profile, where an increase of phosphatdylinositides saturated fatty acids chains and a decrease in C20 fatty acids in cancer cells compared with mammary epithelial cells was observed. At present, information about PL changes in cancer progression is lacking. Therefore, these data will be useful as a starting point to define possible PLs with prospective as biomarkers and disclose metabolic pathways with potential for therapy.

Identification of Biomarkers of Necrosis in Xenografts Using Imaging Mass Spectrometry

“…Importantly, previous publications demonstrated that the analysis of lipid extracts obtained from tumor human cells lines or human cancer biopsies showed increased levels of PC and/or PE ether lipids. In fact, these changes are correlated with malignancy and metastatic potential in various cancers [37][38][39][40][41][42]. Some studies demonstrated that plasmalogens are natural antioxidants, as the ether link is more prone to be oxidized than the double bonds usually found in many phospholipids in their fatty acid moiety [9,43].…”

LXR activation by GW3965 alters fat tissue distribution and adipose tissue inflammation in ob/ob female mice

“…TG and Chol contents were quantifi ed using the colorimetric kits Liquick Cor-TG and Liquick Cor-Chol (Cormay). PL amounts were estimated from total phosphorous content ( 32 ). TG, Chol, and PL amounts were related to tissue weight used for each extract.…”

“…TG, Chol, and PL amounts were related to tissue weight used for each extract. PL classes were separated by thin layer chromatography (TLC) using silica gel plates with concentrating zone 2.5 × 20 cm (Merck) ( 32 ) and identifi ed using standards (Avantis). PLs were extracted with chloroform/methanol (2:1, v/v) for mass spectrometry (MS) analysis.…”

Odróżnienie brodawkowatego raka tarczycy od tkanki nienowotworowej w oparciu o profilowanie lipidów metodą MALDI-MSI

“…Phospholipids, which are the major constituents of cell membranes, play an important role in signal transduction, inflammation, cell proliferation and differentiation [8][9][10], and influence key aspects of cancer phenotype [11,12]. A few papers have reported differences in the lipid profiles of thyroid neoplasms [13][14][15][16], hence this class of compounds could be potentially implemented in classification of thyroid lesions.…”