Department of Hematology, The First Affiliated Hospital of Xi'an Medical University, No. 48, Fenghao West Road, Lianhu District, Xi'an, 710077, China.
Biotechnol Lett. 2021 Jul;43(7):1311-1322. doi: 10.1007/s10529-021-03125-x. Epub 2021 Apr 23.
Bone marrow mesenchymal stem cells (BMMSCs) were proved to play a vital role in multiple myeloma (MM). Polygonatum sibiricum polysaccharide (PSP) was found to have anti-tumor pharmacological effects, yet its interaction with BMMSCs remained poorly understood. Therefore, we explore the effect of PSP on osteogenic differentiation of BMMSCs.
BMMSCs were isolated by density gradient centrifugation. CD90 and CD34 were detected by flow cytometry (FCM). Osteogenic marks were detected by quantitative real-time PCR (qRT-PCR) and Western blotting (WB). The vitality of cells treated with different concentrations of PSP was observed by Cell Counting Kit-8 (CCK-8). ALP staining kit was used to detect the activity of alkaline phosphatase (ALP). Alizarin red staining detected the formation of mineralized nodules. Osteoblast-associated genes were evaluated by qRT-PCR and WB. The phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) signaling pathways were tested by WB.
The BMMSCs showed good growth under an inverted microscope. FCM showed that CD34 and CD45 was low-expressed, whereas CD44, CD90 and CD105 was highly expressed. Compared with the Control group, the expressions of Runx2 and ALP in cells were significantly increased. CCK-8 showed that different concentrations of PSP had no significant effect on the viability of BMMSCs. BMMSCs treated with 25 mg/l PSP were stained the most deeply by ALP. Mineralized nodules in PSP groups dramatically increased, and hit a peak under the action of 25 mg/l PSP. PSP up-regulated p-PI3K, p-AKT, and p-mTOR, but had no significant effect on PI3K, AKT, and mTOR.
PSP induced osteogenic differentiation of BMMSCs from MM patients.
骨髓间充质干细胞（BMMSCs）在多发性骨髓瘤（MM）中起着至关重要的作用。已发现玉竹多糖（PSP）具有抗肿瘤药理作用，但它与 BMMSCs 的相互作用尚不清楚。因此，我们探讨了 PSP 对 BMMSCs 成骨分化的影响。
通过密度梯度离心法分离 BMMSCs。采用流式细胞术（FCM）检测 CD90 和 CD34。通过实时定量 PCR（qRT-PCR）和 Western blot（WB）检测成骨标志物。用细胞计数试剂盒-8（CCK-8）观察不同浓度 PSP 处理的细胞活力。碱性磷酸酶（ALP）染色试剂盒检测 ALP 活性。茜素红染色检测矿化结节的形成。通过 qRT-PCR 和 WB 评估成骨相关基因。通过 WB 检测磷酸肌醇 3-激酶（PI3K）、蛋白激酶 B（AKT）和哺乳动物雷帕霉素靶蛋白（mTOR）信号通路。
倒置显微镜下观察到 BMMSCs 生长良好。FCM 显示 CD34 和 CD45 低表达，而 CD44、CD90 和 CD105 高表达。与对照组相比，细胞中 Runx2 和 ALP 的表达明显增加。CCK-8 显示不同浓度的 PSP 对 BMMSCs 的活力没有显著影响。用 25mg/l PSP 处理的 BMMSCs 的 ALP 染色最深。PSP 组的矿化结节显著增加，在 25mg/l PSP 的作用下达到峰值。PSP 上调了 p-PI3K、p-AKT 和 p-mTOR，但对 PI3K、AKT 和 mTOR 没有显著影响。
PSP 诱导 MM 患者 BMMSCs 成骨分化。
