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J Am Soc Mass Spectrom ; 22(12): 2145-59, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22002228

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.


Subject(s)
Adenocarcinoma/metabolism , Adenocarcinoma/pathology , Brain Neoplasms/metabolism , Brain Neoplasms/secondary , Gangliosides/metabolism , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Microchip Analytical Procedures/methods , Spectrometry, Mass, Electrospray Ionization/methods , Adenocarcinoma of Lung , Aged , Brain Chemistry , Case-Control Studies , Chromatography, Thin Layer , Densitometry , Gangliosides/analysis , Gangliosides/biosynthesis , Gangliosides/chemistry , Humans , Male , Tandem Mass Spectrometry
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