Stromal cells prevent apoptosis of AML cells by up-regulation of anti-apoptotic proteins
Stromal cells prevent apoptosis of AML cells by up-regulation of anti-apoptotic proteins
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DOI：
10.1038/sj.leu.2402608
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发表时间：
2002-09-01
期刊：
影响因子：
11.4
通讯作者：
Andreeff, M
中科院分区：
文献类型：
--
作者：
Konopleva, M;Konoplev, S;Andreeff, M
本研究的目的是研究骨髓基质微环境与白血病细胞之间的相互作用。我们验证了基质细胞通过上调白血病原始细胞中的抗凋亡蛋白来防止AML细胞凋亡的假设。在HL-60和NB-4细胞中，与MS-5基质细胞共培养后，血清剥夺和阿糖胞苷诱导的细胞凋亡明显减少（P < 0.02）。用MS-5细胞的条件培养基再现了这种效果。与基质细胞共培养诱导Bcl-2表达水平，通过PCR分析和流式细胞术。在原发性AML（n = 14）中，与MS-5细胞共培养的细胞中阿糖胞苷诱导的细胞凋亡显著低于对照组（P < 0.001）。当白血病细胞与基质细胞通过微孔插入物分离时，这种效果部分保留（在5/9个样品中，P = 0.04）。间质支持的AML细胞Bcl-2表达水平明显高于对照组（P < 0.01）。在基质层上生长的5/7个样品中Bcl-X-L水平较高。值得注意的是，在诱导化疗耐药的AML患者（n = 6）中，与基质细胞培养后Bcl-2显著增加，但在化疗敏感患者的细胞中没有观察到这种增加。总之，MS-5基质细胞通过调节Bcl-2家族蛋白阻止HL-60细胞和原代AML母细胞的凋亡。在体外基质支持的AML原始细胞中观察到的Bcl-2高表达与体内化疗耐药性的相关性表明，相同的机制可能在体内起作用。
The aim of this study was to study interactions between stromal bone marrow microenvironment and leukemic cells. We tested the hypothesis that stromal cells prevent apoptosis of AML cells by up-regulating anti-apoptotic proteins in leukemic blasts. In HL-60 and NB-4 cells, serum deprivation- and ara-C-induced apoptosis was diminished when cells were cocultured with murine MS-5 stromal cells (P < 0.02). This effect was reproduced with conditioned medium from MS-5 cells. Cocultivation with stromal cells induced Bcl-2 expression levels, both by PCR analysis and flow cytometry. In primary AML (n = 14), ara-C-induced apoptosis was significantly lower in cells cocultured with MS-5 cells than in controls (P < 0.001). This effect was partially preserved when leukemic cells were separated from stromal cells by a microporous insert (in 5/9 samples, P = 0.04). In addition, Bcl-2 levels were significantly higher in stroma-supported than in control CD34(+) AML cells (P < 0.01). Bcl-X-L levels were higher in 5/7 samples grown on stromal layers. Of note, in AML patients resistant to induction chemotherapy (n = 6), Bcl-2 increased significantly after cultivation with stromal cells, but no such increase was noted in cells from chemotherapy-sensitive patients. In conclusion, MS-5 stromal cells prevented apoptosis in HL-60 cells and in primary AML blasts via modulation of Bcl-2 family proteins. The observed association of high Bcl-2 expression in stroma-supported AML blasts in vitro with resistance to chemotherapy in vivo suggests that the same mechanisms may be operational in vivo.