Huwaikem Muneerah A H, Kalamegam Gauthaman, Alrefaei Ghadeer, Ahmed Farid, Kadam Roaa, Qadah Talal, Sait Khalid H W, Pushparaj Peter N
Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, University of Tabuk, Tabuk, Saudi Arabia.
Front Cell Dev Biol. 2021 Mar 18;9:614988. doi: 10.3389/fcell.2021.614988. eCollection 2021.
Emerging resistance to the tyrosine kinase inhibitors that target the BCR-ABL1 oncoprotein has prompted research for novel therapeutics against chronic myeloid leukemia (CML). Herein, we evaluated the tumor inhibitory properties of the human Wharton's jelly stem cells (hWJSCs) co-culture (hWJSC-CC) and their extracts, namely, the hWJSC-conditioned medium (hWJSC-CM; 100%) and hWJSC-lysate (hWJSC-L; 15 μg/ml), on a CML cell line K562 . The hWJSCs expressed mesenchymal stem cell (MSC)-related cluster of differentiation (CD) markers and demonstrated mesodermal tissue differentiation potential. The cell metabolic activity showed a mean maximal decrease in the K562 cells by 49.12, 41.98, and 68.80% following treatment with the hWJSC-CC, hWJSC-CM, and hWJSC-L, respectively, at 72 h. The sub-G1 population in the cell cycle was decreased by 3.2, 4.5, and 3.8% following treatment with the hWJSC-CC, hWJSC-CM, and hWJSC-L, whereas the G2/M cell population was increased by 13.7 and 12.5% with the hWJSC-CM and hWJSC-L, respectively, at 48 h. Annexin V-allophycocyanin (APC) assay showed an increase in the apoptotic cells by 4.0, 3.9, and 4.5% at 48 h. The expression of pro-apoptotic and genes were increased, whereas BIRC5 () was decreased compared with the control. The pro-inflammation-related genes, namely, , , , , , and , were decreased, whereas the anti-inflammatory genes, namely, and , were increased following treatment with the hWJSC-CC, hWJSC-CM, and hWJSC-L at 48 h. Multiplex bead-based cytokine assay also demonstrated decreases in the pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β, IL-6, and IL-12) and an increase in the anti-inflammatory cytokine (IL-10) compared with the control. The pro-inflammatory cytokine IL-8 showed an increase with the hWJSC-CC and decreases with both the hWJSC-CM and the hWJSC-L. The hWJSCs and their extracts inhibited the K562 cells by causing cell cycle arrest and inducing apoptosis the soluble cellular factors. However, an evaluation is necessary to unravel the true potential of the hWJSCs and their extracts before its use in CML inhibition.
对靶向BCR-ABL1癌蛋白的酪氨酸激酶抑制剂出现的耐药性促使人们开展针对慢性粒细胞白血病（CML）的新型疗法研究。在此，我们评估了人脐带华通氏胶间充质干细胞（hWJSCs）共培养物（hWJSC-CC）及其提取物，即hWJSC条件培养基（hWJSC-CM；100%）和hWJSC裂解物（hWJSC-L；15μg/ml）对CML细胞系K562的肿瘤抑制特性。hWJSCs表达间充质干细胞（MSC）相关的分化簇（CD）标志物，并表现出中胚层组织分化潜能。细胞代谢活性显示，在72小时时，用hWJSC-CC、hWJSC-CM和hWJSC-L处理后，K562细胞的平均最大降幅分别为49.12%、41.98%和68.80%。在48小时时，用hWJSC-CC、hWJSC-CM和hWJSC-L处理后，细胞周期中的亚G1期细胞群分别减少了3.2%、4.5%和3.8%，而G2/M期细胞群在用hWJSC-CM和hWJSC-L处理后分别增加了13.7%和12.5%。膜联蛋白V-别藻蓝蛋白（APC）检测显示，在48小时时凋亡细胞增加了4.0%、3.9%和4.5%。与对照组相比，促凋亡基因和的表达增加，而凋亡抑制蛋白5（BIRC5）的表达减少。在用hWJSC-CC.hWJSC-CM和hWJSC-L处理48小时后，促炎相关基因，即、、、、、和的表达减少，而抗炎基因，即和的表达增加。基于多重微珠的细胞因子检测也显示，与对照组相比，促炎细胞因子（IFN-γ、TNF-α、IL-1β、IL-6和IL-12）减少，抗炎细胞因子（IL-10）增加。促炎细胞因子IL-8在用hWJSC-CC处理后增加，在用hWJSC-CM和hWJSC-L处理后均减少。hWJSCs及其提取物通过导致细胞周期停滞和诱导凋亡以及可溶性细胞因子来抑制K562细胞。然而，在将hWJSCs及其提取物用于抑制CML之前，有必要进行进一步评估以揭示其真正潜力。
