HFCL细胞对U937细胞的诱导分化作用

The Differentiation of U937 Cells Induced by Normal Human Bone Marrow Fibroblastoid Stromal Cell Line

为了观察正常人骨髓成纤维样细胞系HFCL对急性单核细胞白血病U937细胞促分化作用,及其对经典诱导分化剂TPA诱导分化作用的影响,先建立U937细胞和HFCL细胞共培养体系,以细胞形态学改变、硝基四氮唑蓝(NBT)、流式细胞仪检测细胞周期和CD11b、CD13、CD14、CD33细胞表面抗原作为诱导分化指标Western印迹检测P38蛋白的表达变化。结果发现,与HFCL细胞共培养后,U937细胞出现分化成熟的形态学改变,且与HFCL细胞直接接触组的诱导分化作用大于用transwell组。同时发现U937细胞与HFCL细胞共培养后,G1期细胞增高,S期细胞减少;CD11b、CD13、CD14和CD33表达增高;且NBT阳性细胞增高至46.3％。Western印迹检测结果显示,直接接触组总P38蛋白表达增加。而且HFCL细胞还能增强TPA对U937的诱导分化作用。

To investigated the effects of normal human bone marrow fibroblastoid stromal cell line (HFCL) on the differentiation of acute myeloid leukemia cell line U937, the coculture system of leukemia cell line U937 and HFCL cells was established at first. Growth curves were detected by cell counting. Cell differentiation was determined by morphologic observation, ability of NBT cells and flow cytometric detection of the expression of CD11b, CD14, CD13 and CD33. Flow cytometry and Western blot were performed to test the changes of cell cycle and expression of P38, respectively. Compared with U937 cells without HFCL cells, the proliferation of U937 cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited, and NBT positive cells increased to 46.3%. The percentage of G1 phase cells of U937 cells with HFCL cells was higher than that without HFCL cells, and the percentage of S phase cells was lower. Meanwhile the expression of CD11b and CD14 increased, however, the expression of CD13 and CD33 didn't change. The expression of P38 in U937 cells with HFCL cells was higher than that in U937 cells without HFCL cells. Meanwhile, HFCL cells could enhance the differentiation of U937 cells induced by TPA. In a word, the normal bone marrow fibroblastoid stromal cells HFCL could induce the differentiation of part of U937 cells into monocyte with the increasing expression of P38 and also enhance the differentiation of U937 cells induced by TPA.