Profiling of N-glycan Alterations in Acute Myeloid Leukemia Cells Before and After Co-cultured with Bone Marrow Derived Stromal Cells

Abstract: Bone marrow derived stromal cells are important components of bone marrow microenvironment (so called niche) where they support hematopoiesis via direct cell-cell interactions with hematopoietic stem/progenitor cells by releasing soluble factors. Glycans， such as N-glycan， are involved in numerous biological processes， including inflammation， cell-cell interactions， morphogenesis， cancer development and progression. In this study， acute myeloid leukemia (AML) cells KG1a co-cultured with bone marrow derived stromal cells HS27a were utilized as the in vitro cell model to study the profiling of N-glycan in KG1a cells before and after stromal contact by using MALDI-TOF-MS (matrix-assistted laser desorption/ionization time of flight mass spectrometry) analysis. Our study showed that the levels of core-fucosylated N-glycans encoded by FUT8 (fucosyltransferase 8)， tri-antennary and tetra-antennary encoded by MGAT5 [mannosyl (α-1，6)-glycoprotein β-1，6-N-acetylglucosaminyltransferase] and MGAT3 (mannosyl-glycoprotein β-1，4-N-acetyl-glucosaminyltransferase) in cocultured KG1a cells were enhanced and their responded genes were also elevated. Meantime， the relative intensity of Man10GlcNAc2Asn N-glycan was down regulated in co-cultured KG1a cells. Consistent with MS results， lectin staining study showed that binding affinity to lectin PHA-E+L (Phaseolus vulgaris Agglutinin) and LCA (Lens culinaris) was enhanced in co-cultured KG1a cells. Profiling the alterations of N-glycan in KG1a cells in present or absent of HS27a cells will further to characterize these significantly differentially expressed N-glycan and their biological functions in bone marrow microenvironment.

CSTR: 32200.14.cjcb.2016.11.0004