复方苦参注射液对多发性骨髓瘤干细胞特性的抑制作用

Compound Kushen injection inhibiting the characteristics of multiple myeloma stem cells

目的探讨复方苦参注射液(compound Kushen injection,CKI)对人多发性骨髓瘤(multiple myeloma,MM)细胞系RPMI-8226中的CD138^(-)细胞及MM干细胞特性的抑制作用。方法采用流式细胞术检测MM细胞中CD138^(-)细胞的比例及不同浓度CKI(0、1、2和4 mg/ml)对CD138^(-)细胞的抑制作用,利用软琼脂克隆形成试验检测不同浓度CKI对MM细胞克隆形成能力的作用,用WB法检测CKI对MM干细胞转录因子Nanog、Oct4表达的影响。结果MM细胞系中存在(3.94±0.71)%的CD138^(-)细胞,CKI对其有抑制作用,不同浓度药物处理组的CD138^(-)细胞比例分别为(3.96±0.76)%、(3.87±0.84)%、(1.72±0.54)%、(0.23±0.11)%,并且可以抑制MM的克隆形成能力,各组的克隆形成数为(69.00±6.56)、(64.33±8.15)、(37.00±8.19)、(16.00±4.58),可降低干细胞转录因子Nanog、Oct4蛋白表达水平。结论CKI能抑制MM中的CD138^(-)细胞及MM干细胞特性,可作为传统抗肿瘤治疗的补充,使MM治愈成为可能,为后期的机制研究及临床研究奠定基础。

Objection To investigate the inhibitory effect of compound Kushen injection(CKI)on CD138^(-) cells in cell line RPMI-8226 and multiple myeloma(MM)stem cells in MM.Methods Flow cytometry was used to detect the proportion of CD138^(-) cells in MM cells and the inhibitory effect of different concentration of CKI(0,1,2 and 4 mg/ml)on CD138^(-) cells.Soft agar assay was performed to examine the influence of colony-formation capacity in MM cells.Western blotting was used to detect the effect of CKI on the expression of stem cell transcription factors Nanog and Oct4 in MM stem cells.Results There was(3.94±0.71)%CD138^(-) cells in MM cell lines.CKI had inhibitive effect on CD138^(-)cells,and the proportion of CD138^(-) cells treated by different concentration of CKI were(3.96±0.76)%,(3.87±0.84)%,(1.72±0.54)%and(0.23±0.11)%,respectively.CKI could restrain the colony formation ability of MM cells,the colony formation number in different groups were(69.00±6.56),(64.33±8.15),(37.00±8.19)and(16.00±4.58).CKI could reduce the expression of stem cell transcription factors Nanog and Oct4.Conclusions CKI can inhibit CD138^(-) cells and MM stem cell properties in MM,which can be used as a supplement to traditional anti-tumor therapies,making it possible to cure MM,and laying a foundation for later mechanism research and clinical study.