Background

The osteoblast differentiation of bone marrow-derived stem cells (BMSCs) is impaired in multiple myeloma (MM). We investigated the effects of sodium copper chlorophyllin (SCC) on osteoblast differentiation ability of BMSCs from MM.

Methods

Clinical bone marrow samples were collected. Fluorescence Activated Cell Sorter (FACS) was used to identify surface markers of BMSCs. BMSCs were treated with different concentrations of SCC and cell viability was detected by MTT assay. Relative mRNA and protein expressions of transforming growth factor-β1 (TGF-β1), SMAD2/3, osteogenic differentiation indicators (RUNX2 and OCN) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Alkaline phosphatase (ALP) was stained for activity detection. Formation of calcium nodus of BMSCs was examined by Alizarin Red S staining.

Results

CD90 and CD105 were high-expressed, but CD34 and CD45 were not expressed in BMSCs. BMSCs in MM group showed a lower expression of TGF-β1 and a lower degree of osteogenic differentiation. SCC enhanced activities of BMSCs, ALP activity, and formation of calcium nodus, activated TGF-β1, SMAD2/3 pathway and increased RUNX2 and OCN expressions in BMSCs. Silencing TGF-β1 reversed the effects of SCC on BMSCs in MM.

Conclusion

SCC could effectively improve the proliferation and osteogenic differentiation of BMSCs in MM through regulating TGF-β1.

中文翻译：

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叶绿素铜钠通过调节TGF-β1对多发性骨髓瘤骨髓间充质干细胞功能障碍的有效作用。

背景

骨髓源性干细胞 (BMSC) 的成骨细胞分化在多发性骨髓瘤 (MM) 中受损。我们研究了叶绿素铜钠 (SCC) 对 MM 的 BMSCs 成骨细胞分化能力的影响。

方法

收集临床骨髓样本。荧光激活细胞分选仪 (FACS) 用于鉴定 BMSCs 的表面标志物。用不同浓度的SCC处理BMSCs，MTT法检测细胞活力。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质印迹法测量转化生长因子-β1（TGF-β1）、SMAD2/3、成骨分化指标（RUNX2和OCN）的相对mRNA和蛋白表达。碱性磷酸酶 (ALP) 染色用于活性检测。茜素红 S 染色检查 BMSCs 钙结节的形成。

结果

CD90 和 CD105 高表达，但 CD34 和 CD45 在 BMSC 中不表达。MM组骨髓间充质干细胞TGF-β1表达较低，成骨分化程度较低。SCC 增强 BMSCs 的活性、ALP 活性和钙结节的形成，激活 TGF-β1、SMAD2/3 通路并增加 BMSCs 中 RUNX2 和 OCN 的表达。沉默 TGF-β1 逆转了 SCC 对 MM 中 BMSC 的影响。

结论

SCC可通过调节TGF-β1有效促进MM中BMSCs的增殖和成骨分化。