Objective: To investigate the effect of bone marrow stromal cell glycosyltransferase B4GALT1 expression on hematopoietic cell proliferation and its upstream regulation mechanism.
Methods: B4GALT1 was overexpressed in human bone marrow stromal cell line HS5, which was then co-cultured with acute myeloid leukemia cell line KG1a. And its effect on hematopoietic cell proliferation was detected by flow cytometry. Dual luciferase reporter assay, real-time PCR and Western blot were used to predict and validate upstream transcription factors that regulate stromal cell B4GALT1 expression.
Results: Overexpression of B4GALT1 in HS5 significantly promoted the proliferation of KG1a in the co-culture system. B4GALT1 expression in stromal cells positively correlated with upstream c-Jun expression, which was verified by JNK/c-Jun inhibitors.
Conclusion: The differential expression of glycosyltransferases and their corresponding glycosylation in the hematopoietic microenvironment play an important role.
题目: 骨髓基质细胞中B4GALT1对与其共培养的人急性髓系白血病细胞株增殖的影响.
目的: 探讨骨髓基质细胞糖基转移酶B4GALT1的表达对造血细胞增殖的影响及其上游调控机制.
方法: 在人骨髓来源基质细胞HS5中过表达B4GALT1，并与急性髓系白血病细胞KG1a共培养，采用流式细胞术检测其对造血细胞增殖的影响；应用双荧光素酶报告基因检测、实时荧光定量PCR和蛋白免疫印迹法对调控基质细胞B4GALT1表达的上游转录因子进行预测和验证.
结果: HS5中过表达B4GALT1能显著促进共培养体系中KG1a的增殖。基质细胞中B4GALT1表达与上游c-Jun表达呈正相关，并通过JNK/c-Jun抑制剂进行了验证.
结论: 糖基转移酶及其引起的相应糖基化差异在造血微环境中起到的调控作用，其作用机制有待更深入的研究.