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骨髓基质细胞是骨髓微环境的重要组成部分,在促进造血干细胞生成血细胞的各个环节中起重要作用。作为细胞结构重要组成部分的糖链(如N-糖链),广泛参与了细胞之间的黏附、分化、增殖及信号转导等过程。为了探究骨髓微环境对血液细胞表面N-糖链变化的影响,该研究使用人骨髓来源的基质细胞HS27a与急性髓系白血病细胞KG1a共培养的体外模型,利用基质辅助激光解析电离飞行时间质谱(matrix-assistted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)技术从糖组学角度比较了与骨髓基质细胞共培养前后的KG1a细胞中N-糖链变化,并结合实时荧光定量PCR(Real-time PCR)和细胞凝集素染色对质谱结果加以验证。结果显示,与HS27a共培养后的KG1a中高甘露糖型Man10Glc NAc2Asn结构N-糖链相对强度降低,平分型、四天线复杂型和岩藻糖化的N-糖链相对强度增加,合成相应糖链的MGAT5[mannosyl(α-1,6)-glycoproteinβ-1,6-N-acetyl-glucosaminyltransferase]、MGAT3(mannosyl-glycoproteinβ-1,4-N-acetylglucosaminyltransferase)和FUT8(fucosyltransferase 8)基因水平表达上调。细胞染色结果证明,共培养后的KG1a对凝集素PHA-E+L(Phaseolus vulgaris Agglutinin)和LCA(Lens culinaris)的亲和作用增强。该研究从糖生物学角度揭示了骨髓来源的基质细胞对急性髓系白血病细胞在N-糖链变化上的影响。未来的研究重点将放在共培养前后急性髓系白血病细胞中显著差异糖链的生物学功能上,以了解N-糖链在骨髓微环境中所发挥的重要作用。
Bone marrow stromal cells are an important part of the bone marrow microenvironment and play an important role in promoting hematopoietic stem cells to produce blood cells in all aspects. As an important part of the cellular structure of the sugar chain (such as N-sugar chain), widely involved in cell adhesion, differentiation, proliferation and signal transduction process. In order to explore the effects of bone marrow microenvironment on the changes of N-glycan on blood cells, the in vitro model of human bone marrow-derived stromal cells HS27a co-cultured with acute myeloid leukemia cells KG1a was established by matrix-assisted laser desorption / ionization time of flight mass spectrometry The changes of N-glycans in KG1a cells before and after co-culture with bone marrow stromal cells were compared by using the matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS) (Real-time PCR) and cytokinin staining mass spectrometry results to be verified. The results showed that the relative intensity of N-glycan of Man10Glc NAc2Asn with high Mannose type in KG1a after co-culture with HS27a decreased, and the relative intensities of bisected, four-antenna complex and fucosylated N-glycan increased, Of MGAT5 [mannosyl (α-1,6) -glycoproteinβ-1,6-N-acetyl-glucosaminyltransferase] and MGAT3 (mannosyl-glycoproteinβ-1,4-N-acetylglucosaminyltransferase) and FUT8 (fucosyltransferase 8) The results of cell staining showed that the co-cultured KG1a had an enhanced affinity for PHA-E + L (Phaseolus vulgaris Agglutinin) and LCA (Lens culinaris). This study revealed the effect of bone marrow-derived stromal cells on N-glycan change in acute myeloid leukemia cells from the perspective of glycogen biology. Future research will focus on the biological function of significantly different glycan chains in acute myeloid leukemia cells before and after co-culture in order to understand the important role that N-glycans play in the bone marrow microenvironment.