Janfada Mehrnaz, Vahdat Sadaf, Kaviani Saeid
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Applied Cell Sciences Division, Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Adv Pharm Bull. 2025 Apr 24;15(2):440-452. doi: 10.34172/apb.025.42774. eCollection 2025 Jul.
The survival and progression of multiple myeloma (MM) cells rely heavily on supportive factors and cells within the MM microenvironment, notably macrophages. The PI3K signaling pathway plays a crucial role in both myeloma cells survival and macrophage polarity, making it a potential target for altering the MM microenvironment dynamics.
In this study, the impact of LY294002, a PI3K signaling pathway inhibitor, on the viability of U266 myeloma cells in mono-culture and MM patient-derived bone marrow mononuclear cells (BM-MNCs) in co-culture was investigated. Additionally, the effect of treatments on the M1/M2 macrophage ratio was assessed. Cultures were conducted in both two-dimensional (2D) matrix-free and fibrin gel-based three-dimensional (3D) environments.
The treatment significantly increased U266 cell death in 2D cultures, dose-dependently compared to control. However, this effect was not replicated in 3D cultures. In both 2D and 3D cultures, the percentages of cells in G0/G1 phase were dose-dependently increased, compared to the untreated control. However, the percentages of cells in S and G2/M phases in both 2D and 3D cultures were dose-dependently decreased, compared to control. Treatment of BM-MNCs with LY294002 showed patient- and culture-dependent patterns of CD138 myeloma cell death and M1/M2 macrophage ratio, contrasting the observed consistent responses in U266 mono-culture.
LY294002 affected U266 cell viability and cell cycle in a dose-dependent manner in 2D mono-cultures. However, its impact varied in 3D cultures. Treatment of MNCs showed varied responses based on individuals and culture conditions, underscoring the need for more similar tumor microenvironment (TME) recapitulation for drug screening.
多发性骨髓瘤（MM）细胞的存活和进展严重依赖于MM微环境中的支持因子和细胞，尤其是巨噬细胞。PI3K信号通路在骨髓瘤细胞存活和巨噬细胞极性方面均发挥关键作用，使其成为改变MM微环境动态的潜在靶点。
在本研究中，研究了PI3K信号通路抑制剂LY294002对单培养的U266骨髓瘤细胞以及共培养的MM患者来源的骨髓单个核细胞（BM-MNCs）活力的影响。此外，评估了处理对M1/M2巨噬细胞比例的影响。培养在二维（2D）无基质和基于纤维蛋白凝胶的三维（3D）环境中进行。
在2D培养中，与对照组相比，该处理显著增加了U266细胞死亡，且呈剂量依赖性。然而，这种效应在3D培养中未得到重现。在2D和3D培养中，与未处理的对照组相比，G0/G1期细胞的百分比均呈剂量依赖性增加。然而，与对照组相比，2D和3D培养中S期和G2/M期细胞的百分比均呈剂量依赖性降低。用LY294002处理BM-MNCs显示出CD138骨髓瘤细胞死亡和M1/M2巨噬细胞比例的患者和培养依赖性模式，这与在U266单培养中观察到的一致反应形成对比。
LY294002在2D单培养中以剂量依赖性方式影响U266细胞活力和细胞周期。然而，其在3D培养中的影响有所不同。对单个核细胞的处理显示出基于个体和培养条件的不同反应，强调了在药物筛选中需要更多类似的肿瘤微环境（TME）重现。
