Marcus H, Attar-Schneider O, Dabbah M, Zismanov V, Tartakover-Matalon S, Lishner M, Drucker L
Oncogenetic Laboratory, Tel Aviv University, Tel Aviv, Israel; Sackler faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Oncogenetic Laboratory, Tel Aviv University, Tel Aviv, Israel; Internal Medicine Department, Meir Medical Center, Kfar Saba, Tel Aviv University, Tel Aviv, Israel; Sackler faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Cell Signal. 2016 Jun;28(6):620-30. doi: 10.1016/j.cellsig.2016.03.003. Epub 2016 Mar 11.
Bone marrow mesenchymal stem cells' (BM-MSCs) role in multiple myeloma (MM) pathogenesis is recognized. Recently, we have published that co-culture of MM cell lines with BM-MSCs results in mutual modulation of phenotype and proteome (via translation initiation (TI) factors eIF4E/eIF4GI) and that there are differences between normal donor BM-MSCs (ND-MSCs) and MM BM-MSCs (MM-MSCs) in this crosstalk. Here, we aimed to assess the involvement of soluble BM-MSCs' (ND, MM) components, more easily targeted, in manipulation of MM cell lines phenotype and TI with specific focus on microvesicles (MVs) capable of transferring critical biological material. We applied ND and MM-MSCs 72h secretomes to MM cell lines (U266 and ARP-1) for 12-72h and then assayed the cells' (viability, cell count, cell death, proliferation, cell cycle, autophagy) and TI (factors: eIF4E, teIF4GI; regulators: mTOR, MNK1/2, 4EBP; targets: cyclin D1, NFκB, SMAD5, cMyc, HIF1α). Furthermore, we dissected the secretome into >100kDa and <100kDa fractions and repeated the experiments. Finally, MVs were isolated from the ND and MM-MSCs secretomes and applied to MM cell lines. Phenotype and TI were assessed. Secretomes of BM-MSCs (ND, MM) significantly stimulated MM cell lines' TI, autophagy and proliferation. The dissected secretome yielded different effects on MM cell lines phenotype and TI according to fraction (>100kDa- repressed; <100kDa- stimulated) but with no association to source (ND, MM). Finally, in analyses of MVs extracted from BM-MSCs (ND, MM) we witnessed differences in accordance with source: ND-MSCs MVs inhibited proliferation, autophagy and TI whereas MM-MSCs MVs stimulated them. These observations highlight the very complex communication between MM and BM-MSCs and underscore its significance to major processes in the malignant cells. Studies into the influential MVs cargo are underway and expected to uncover targetable signals in the regulation of the TI/proliferation/autophagy cascade.
骨髓间充质干细胞（BM-MSCs）在多发性骨髓瘤（MM）发病机制中的作用已得到认可。最近，我们发表了关于MM细胞系与BM-MSCs共培养导致表型和蛋白质组相互调节（通过翻译起始（TI）因子eIF4E/eIF4GI）的研究，并且在这种相互作用中，正常供体BM-MSCs（ND-MSCs）和MM BM-MSCs（MM-MSCs）之间存在差异。在此，我们旨在评估更易于靶向的可溶性BM-MSCs（ND、MM）成分在操纵MM细胞系表型和TI中的作用，特别关注能够传递关键生物物质的微泡（MVs）。我们将ND和MM-MSCs 72小时的分泌产物应用于MM细胞系（U266和ARP-1）12至72小时，然后检测细胞的（活力、细胞计数、细胞死亡、增殖、细胞周期、自噬）和TI（因子：eIF4E、teIF4GI；调节因子：mTOR、MNK1/2、4EBP；靶点：细胞周期蛋白D1、NFκB、SMAD5、cMyc、HIF1α）。此外，我们将分泌产物分离为大于100kDa和小于100kDa的组分，并重复实验。最后，从ND和MM-MSCs分泌产物中分离出MVs并应用于MM细胞系。评估表型和TI。BM-MSCs（ND、MM）的分泌产物显著刺激MM细胞系的TI、自噬和增殖。根据组分（大于100kDa - 抑制；小于100kDa - 刺激），分离后的分泌产物对MM细胞系表型和TI产生不同影响，但与来源（ND、MM）无关。最后，在对从BM-MSCs（ND、MM）中提取的MVs的分析中，我们发现根据来源存在差异：ND-MSCs的MVs抑制增殖、自噬和TI，而MM-MSCs的MVs则刺激它们。这些观察结果突出了MM与BM-MSCs之间非常复杂的通讯，并强调了其对恶性细胞主要过程的重要性。对有影响的MVs货物的研究正在进行中，预计将揭示TI/增殖/自噬级联调节中的可靶向信号。
