Ding Jiang-Hua, Yang Shao-Lin, Zhu Shu-Lang
Department of Hematology & Oncology, Jiujiang University Clinical Medical College & Affiliated Hospital, Jiujiang 332000, Jiangxi Province, China.
Department of Pharmacy, Jiujiang University Clinical Medical College & Affiliated Hospital, Jiujiang 332000, Jiangxi Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb;31(1):170-178. doi: 10.19746/j.cnki.issn.1009-2137.2023.01.027.
To investigate the expression of pyruvate kinase M2 (PKM2) in bone marrow mesenchymal stem cells (BMSCs) in myeloma bone disease (MBD) and its effect on osteogenic and adipogenic differentiation of BMSCs.
BMSCs were isolated from bone marrow of five patients with multiple myeloma (MM) (MM group) and five with iron deficiency anemia (control group) for culture and identification. The expression of PKM2 protein were compared between the two groups. The differences between osteogenic and adipogenic differentiation of BMSCs were assessed by using alkaline phosphatase (ALP) and oil red O staining, and detecting marker genes of osteogenesis and adipogenesis. The effect of MM cell line (RPMI-8226) and BMSCs co-culture on the expression of PKM2 was explored. Functional analysis was performed to investigate the correlations of PKM2 expression of MM-derived BMSCs with osteogenic and adipogenic differentiation by employing PKM2 activator and inhibitor. The role of orlistat was explored in regulating PKM2 expression, osteogenic and adipogenic differentiation of MM-derived BMSCs.
Compared with control, MM-originated BMSCs possessed the ability of increased adipogenic and decreased osteogenic differentiation, and higher level of PKM2 protein. Co-culture of MM cells with BMSCs markedly up-regulated the expression of PKM2 of BMSCs. Up-regulation of PKM2 expression could promote adipogenic differentiation and inhibit osteogenic differentiation of MM-derived BMSCs, while down-regulation of PKM2 showed opposite effect. Orlistat significantly promoted osteogenic differentiation in MM-derived BMSCs via inhibiting the expression of PKM2.
The overexpression of PKM2 can induce the inhibition of osteogenic differentiation of BMSCs in MBD. Orlistat can promote the osteogenic differentiation of BMSCs via inhibiting the expression of PKM2, indicating a potential novel agent of anti-MBD therapy.
探讨丙酮酸激酶M2(PKM2)在骨髓瘤骨病(MBD)骨髓间充质干细胞(BMSCs)中的表达及其对BMSCs成骨和成脂分化的影响。
从5例多发性骨髓瘤(MM)患者(MM组)和5例缺铁性贫血患者(对照组)的骨髓中分离BMSCs进行培养和鉴定。比较两组PKM2蛋白的表达。采用碱性磷酸酶(ALP)和油红O染色以及检测成骨和成脂标记基因来评估BMSCs成骨和成脂分化的差异。探讨MM细胞系(RPMI-8226)与BMSCs共培养对PKM2表达的影响。通过使用PKM2激活剂和抑制剂进行功能分析,以研究MM来源的BMSCs中PKM2表达与成骨和成脂分化的相关性。探讨奥利司他在调节MM来源的BMSCs中PKM2表达、成骨和成脂分化中的作用。
与对照组相比,MM来源的BMSCs具有增强的成脂能力和降低的成骨分化能力,且PKM2蛋白水平更高。MM细胞与BMSCs共培养显著上调了BMSCs中PKM2的表达。PKM2表达上调可促进MM来源的BMSCs的成脂分化并抑制其成骨分化,而PKM2表达下调则显示相反的作用。奥利司他通过抑制PKM2的表达显著促进了MM来源的BMSCs的成骨分化。
PKM2的过表达可诱导MBD中BMSCs成骨分化的抑制。奥利司他可通过抑制PKM2的表达促进BMSCs的成骨分化,表明其可能是抗MBD治疗的新型药物。