Method for Lipidomic Analysis:  p53 Expression Modulation of Sulfatide, Ganglioside, and Phospholipid Composition of U87 MG Glioblastoma Cells

Abstract

Glioblastoma multiforme (GBM) is the most common and malignant form of primary brain tumors. It is highly invasive and current treatment options have not improved the survival rate over the past twenty years. Novel approaches and technologies from systems biology have the potential to identify biomarkers that could serve as new therapeutic targets for GBM. This study employed

Targeted analysis of ganglioside and sulfatide molecular species by LC/ESI-MS/MS with theoretically expanded multiple reaction monitoring

“…Electrospray is considered a particularly useful ionization method in MS because of soft ionization with the capability of easily connecting with HPLC (15)(16)(17)(18)(19)(20)(21)(22). Individual molecular species of gangliosides can be separately eluted in reverse-phase HPLC by their different sugar chain lengths, especially the number of sialic acid residues; they can also be separately eluted by their fatty alcohol or N-acyl carbon chain lengths.…”

Overexpression of ST6GalNAcV, a ganglioside-specific α2,6-sialyltransferase, inhibits glioma growth in vivo

“…We compared polar lipid changes in control U373MG cells and the two highest expressing ST6GalNAc transfectants (S8 and S25) by use of a detailed, sensitive, semiquantitative method using lipid extraction, one-step nano-liquid chromatography (nLC) separation, and high-resolution mass analysis (18,19). We observed significant modulation of specific gangliosides in the α-series pathway (Fig.…”

“…Polar lipids were extracted from cells and analyzed by nLC-MS as described (18,19) and detailed in SI Methods. Datadependent MS/MS was performed in the linear quadrupole ion trap (CID) during collection of the ICR time-domain data.…”

Role of sulfatide in normal and pathological cells and tissues

“…Although CST and CGT appear to recognize to some extent a wide spectrum of hydrophobic ceramide moieties, sulfatide and GalCer appear to be preferentially synthesized from ceramides with C22 and C24 FAs in rat organs and human serous papillary ovarian carcinoma. In addition, for the human glioma cell line U87 MG, overexpression of p53 results in a dramatically increased level of sulfatide possessing C18, despite no signifi cant change in the level of sulfatide possessing C24 ( 9 ). Genes other than CST and CGT can also have an effect on ceramide composition of sulfatide.…”