title: Assessment of the molecular expression and structure of gangliosides in brain metastasis of lung adenocarcinoma by an advanced approach based on fully automated chip-nanoelectrospray mass spectrometry.
abstract: Gangliosides (GGs), sialic acid-containing glycosphingolipids, are known to be involved in the invasive/metastatic behavior of brain tumor cells. Development of modern methods for determination of the variations in GG expression and structure during neoplastic cell transformation is a priority in the field of biomedical analysis. In this context, we report here on the first optimization and application of chip-based nanoelectrospray (NanoMate robot) mass spectrometry (MS) for the investigation of gangliosides in a secondary brain tumor. In our work a native GG mixture extracted and purified from brain metastasis of lung adenocarcinoma was screened by NanoMate robot coupled to a quadrupole time-of-flight MS. A native GG mixture from an age-matched healthy brain tissue, sampled and analyzed under identical conditions, served as a control. Comparative MS analysis demonstrated an evident dissimilarity in GG expression in the two tissue types. Brain metastasis is characterized by many species having a reduced N-acetylneuraminic acid (Neu5Ac) content, however, modified by fucosylation or O-acetylation such as Fuc-GM4, Fuc-GM3, di-O-Ac-GM1, O-Ac-GM3. In contrast, healthy brain tissue is dominated by longer structures exhibiting from mono- to hexasialylated sugar chains. Also, significant differences in ceramide composition were discovered. By tandem MS using collision-induced dissociation at low energies, brain metastasis-associated GD3 (d18:1/18:0) species as well as an uncommon Fuc-GM1 (d18:1/18:0) detected in the normal brain tissue could be structurally characterized. The novel protocol was able to provide a reliable compositional and structural characterization with high analysis pace and at a sensitivity situated in the fmol range.
authors: Department of Chemical and Biological Sciences, Aurel Vlaicu University of Arad, Revolutiei Blvd. 77, RO-310130, Arad, Romania. alina.zamfir@uav.ro
authors: Alina D
authors: AD
authors: Zamfir
authors: Alina
authors: A
authors: Serb
authors: Željka
authors: Ž
authors: Vukeli
authors: Corina
authors: C
authors: Flangea
authors: Catalin
authors: C
authors: Schiopu
authors: Dragana
authors: D
authors: Fabris
authors: Svjetlana
authors: S
authors: Kalanj-Bognar
authors: Florina
authors: F
authors: Capitan
authors: Eugen
authors: E
authors: Sisu
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology*; D000077192:Adenocarcinoma of Lung; D000368:Aged; D001923:Brain Chemistry; D001932:Brain Neoplasms / Q000378:metabolism* / Q000556:secondary*; D016022:Case-Control Studies; D002855:Chromatography, Thin Layer; D003720:Densitometry; D005732:Gangliosides / Q000032:analysis / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D046208:Microchip Analytical Procedures / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D005732:Gangliosides