a Department of Proteomics , Institute of Hematology and Blood Transfusion , U Nemocnice 1, Prague , Czech Republic.
Cell Adh Migr. 2018 May 4;12(3):286-298. doi: 10.1080/19336918.2017.1344796. Epub 2017 Aug 18.
Interaction of leukemia blasts with the bone marrow extracellular matrix often results in protection of leukemia cells from chemotherapy and in persistence of the residual disease which is on the basis of subsequent relapses. The adhesion signaling pathways have been extensively studied in adherent cells as well as in mature haematopoietic cells, but the adhesion structures and signaling in haematopoietic stem and progenitor cells, either normal or malignant, are much less explored. We analyzed the interaction of leukemia cells with fibronectin (FN) using interference reflection microscopy, immunofluorescence, measurement of adherent cell fraction, real-time microimpedance measurement and live cell imaging. We found that leukemia cells form very dynamic adhesion structures similar to early stages of focal adhesions. In contrast to adherent cells, where Src family kinases (SFK) belong to important regulators of focal adhesion dynamics, we observed only minor effects of SFK inhibitor dasatinib on leukemia cell binding to FN. The relatively weak involvement of SFK in adhesion structure regulation might be associated with the lack of cytoskeletal mechanical tension in leukemia cells. On the other hand, active Lyn kinase was found to specifically localize to leukemia cell adhesion structures and a less firm cell attachment to FN was often associated with higher Lyn activity (this unexpectedly occurred also after cell treatment with the inhibitor SKI-1). Lyn thus may be important for signaling from integrin-associated complexes to other processes in leukemia cells.
白血病细胞与骨髓细胞外基质的相互作用常常导致白血病细胞免受化疗的影响,并导致残留疾病的持续存在,这是随后复发的基础。粘附信号通路在粘附细胞以及成熟造血细胞中得到了广泛研究,但在正常或恶性造血干细胞和祖细胞中的粘附结构和信号通路的研究要少得多。我们使用干涉反射显微镜、免疫荧光、粘附细胞分数测量、实时微阻抗测量和活细胞成像分析了白血病细胞与纤维连接蛋白(FN)的相互作用。我们发现白血病细胞形成非常动态的粘附结构,类似于早期的粘着斑。与粘附细胞不同,粘着斑动力学的重要调节因子属于 Src 家族激酶(SFK),我们观察到 SFK 抑制剂 dasatinib 对白血病细胞与 FN 的结合只有很小的影响。SFK 在粘附结构调节中的相对较弱作用可能与白血病细胞中缺乏细胞骨架机械张力有关。另一方面,我们发现活性 Lyn 激酶特异性定位于白血病细胞粘附结构,而 FN 上更不牢固的细胞附着通常与更高的 Lyn 活性相关(即使在细胞用抑制剂 SKI-1 处理后,这种情况也出人意料地发生)。因此,Lyn 可能对整合素相关复合物向白血病细胞中的其他过程传递信号很重要。