Induction of programmed cell death in human hematopoietic cell lines by fibronectin via its interaction with very late antigen 5.
Induction of programmed cell death in human hematopoietic cell lines by fibronectin via its interaction with very late antigen 5.
复制标题
纤连蛋白与非常晚的抗原5通过纤连蛋白的相互作用诱导人造血细胞系中的程序性细胞死亡。
DOI：
10.1084/jem.179.6.1757
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发表时间：
1994-06-01
影响因子：
15.3
通讯作者：
Kanayama, Y
中科院分区：
文献类型：
--
作者：
Sugahara, H;Kanakura, Y;Furitsu, T;Ishihara, K;Oritani, K;Ikeda, H;Kitayama, H;Ishikawa, J;Hashimoto, K;Kanayama, Y
纤连蛋白 (FN)、胶原蛋白和层粘连蛋白等细胞外基质 (ECM) 分子在造血过程中发挥重要作用。然而，人们对ECM分子调节人类造血祖细胞增殖的精确机制知之甚少。在这项研究中，我们研究了 ECM 分子，特别是 FN，对髓系白血病细胞系 M07E 增殖的影响，该细胞系响应人粒细胞/巨噬细胞集落刺激因子 (GM-CSF) 或干细胞因子 (SCF) 进行增殖。 [3H]胸苷掺入和细胞计数测定表明，FN 以剂量依赖性方式显着抑制 GM-CSF 或 SCF 诱导的 M07E 细胞增殖，而 I 型和 IV 型胶原蛋白几乎没有或没有诱导抑制。 M07E 细胞的生长抑制并非由于 FN 对配体结合的抑制作用或来自 GM-CSF 或 SCF 受体的信号转导途径中的非常早期的事件所致。碘化丙啶染色后使用流式细胞术进行DNA含量分析表明，用FN处理M07E细胞并没有阻止细胞在用GM-CSF或SCF刺激后进入细胞周期，而该处理导致亚二倍体峰的出现。此外，即使在 GM-CSF 或 SCF 存在的情况下，FN 也能诱导细胞内寡核小体 DNA 断裂和染色质浓缩，这表明程序性细胞死亡（细胞凋亡）参与了 FN 诱导的生长抑制。 FN 诱导的生长抑制或凋亡可通过添加抗 FN 抗体、抗极晚期抗原 5 单克隆抗体（抗 VLA5 mAb）或 GRGDSP 肽来挽救，但不能通过添加抗 VLA4 mAb 或 GRGESP 肽来挽救，这表明 FN 对 M07E 细胞的作用是通过 VLA5 介导的。此外，在除M07E细胞之外的VLA5阳性人造血细胞系中可检测到FN诱导的细胞凋亡，但在任何VLA5阴性细胞系中均未检测到。这些结果表明，FN 能够通过其与 VLA5 的相互作用诱导细胞凋亡，并且还提出了 FN-VLA5 相互作用可能至少部分地促进造血负向调节的可能性。
Extracellular matrix (ECM) molecules such as fibronectin (FN), collagens, and laminin have important roles in hematopoiesis. However, little is known about the precise mechanisms by which ECM molecules regulate proliferation of human hematopoietic progenitor cells. In this study, we have investigated the effects of ECM molecules, particularly of FN, on the proliferation of a myeloid leukemia cell line, M07E, which proliferates in response to either human granulocyte/macrophage colony-stimulating factor (GM-CSF) or stem cell factor (SCF). The [3H]thymidine incorporation and cell enumeration assays showed that FN strikingly inhibited GM-CSF- or SCF-induced proliferation of M07E cells in a dose-dependent manner, whereas little or no inhibition was induced by collagen types I and IV. The growth suppression of M07E cells was not due to the inhibitory effect of FN on ligand binding or very early events in the signal transduction pathways from the GM-CSF or SCF receptors. DNA content analysis using flow cytometry after staining with propidium iodide revealed that the treatment of M07E cells with FN did not block the entry of the cells into the cell cycle after stimulation with GM-CSF or SCF, whereas the treatment resulted in the appearance of subdiploid peak. Furthermore, FN was found to induce oligonucleosomal DNA fragmentation and chromatin condensation in the cells even in the presence of GM-CSF or SCF, suggesting the involvement of programmed cell death (apoptosis) in the FN-induced growth suppression. The growth suppression or apoptosis induced by FN was rescued by the addition of either anti-FN antibody, anti-very late antigen 5 monoclonal antibody (anti-VLA5 mAb), or GRGDSP peptide, but not by that of anti-VLA4 mAb or GRGESP peptide, suggesting that the FN effects on M07E cells were mediated through VLA5. In addition, the FN- induced apoptosis was detectable in VLA5-positive human hematopoietic cell lines other than M07E cells, but not in any of the VLA5-negative cell lines. These results suggest that FN is capable of inducing apoptosis via its interaction with VLA5, and also raise the possibility that the FN-VLA5 interaction may contribute, at least in part, to negative regulation of hematopoiesis.