Farahzadi Raheleh, Fathi Ezzatollah, Vandghanooni Somayeh, Valipour Behnaz
Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.
Regen Ther. 2024 Aug 28;26:646-653. doi: 10.1016/j.reth.2024.08.008. eCollection 2024 Jun.
The effect of mesenchymal stem cells (MSCs) on the immortal characteristics of malignant cells, particularly hematologic cancer cells, remains a topic of debate, with the underlying mechanisms still requiring further elucidation. We explored the effect of the bone marrow-derived MSCs (BM-MSCs) on CD34 leukemic stem cells (LSCs) enriched from the KG1-a cell line by assessing apoptosis, measuring cytokine levels, and examining TERT protein expression. Additionally, the potential signaling pathways implicated in this process, such as P53, PTEN, NF-κB, ERK1/2, Raf-1, and H-RAS, were also investigated.
CD34 LSCs were enriched from the KG1-a cell line with the magnetic activated cell sorting (MACS) method. Two cell populations (BM-MSCs and CD34 LSCs) were co-cultured on trans well plates for seven days. Next, CD34 LSCs were collected and subjected to Annexin V/PI assay, cytokine measurement, and western blotting.
BM-MSCs caused a significant increase in early and late apoptosis in the CD34LSCs. The significant presence of interleukin (IL)-2 and IL-4 was evident in the co-cultured media. In addition, BM-MSCs significantly increased the protein expression of P53, PTEN, NF-κB, and significantly decreased p-ERK1/2, Raf-1, H-RAS, and TERT.
The mentioned effects of IL-2 and IL-4 cytokines released from BM-MSCs on CD34 LSCs as therapeutic agents were applied by the components of P53, PTEN, NF-κB, p-ERK1/2, Raf-1, and H-RAS signaling pathways.
间充质干细胞（MSCs）对恶性细胞，尤其是血液系统癌细胞的永生特性的影响仍是一个有争议的话题，其潜在机制仍需进一步阐明。我们通过评估细胞凋亡、测量细胞因子水平和检测端粒酶逆转录酶（TERT）蛋白表达，探讨了骨髓来源的间充质干细胞（BM-MSCs）对从KG1-a细胞系富集的CD34白血病干细胞（LSCs）的影响。此外，还研究了参与这一过程的潜在信号通路，如P53、PTEN、核因子κB（NF-κB）、细胞外信号调节激酶1/2（ERK1/2）、 Raf-1和H-RAS。
采用磁珠分选法从KG1-a细胞系中富集CD34 LSCs。将两个细胞群体（BM-MSCs和CD34 LSCs）在Transwell板上共培养7天。接下来，收集CD34 LSCs并进行膜联蛋白V/碘化丙啶（Annexin V/PI）检测、细胞因子测定和蛋白质免疫印迹法检测。
BM-MSCs导致CD34 LSCs早期和晚期凋亡显著增加。共培养培养基中白细胞介素（IL）-2和IL-4显著存在。此外，BM-MSCs显著增加了P53、PTEN、NF-κB的蛋白表达，并显著降低了磷酸化ERK1/2、Raf-1、H-RAS和TERT的表达。
BM-MSCs释放的IL-2和IL-4细胞因子对CD34 LSCs的上述作用是通过P53、PTEN、NF-κB、磷酸化ERK1/2、Raf-1和H-RAS信号通路的成分发挥治疗作用的。
