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目的观察人脐血基质细胞及骨髓瘤患者骨髓基质细胞对骨髓瘤KM3细胞生物学特性的作用。方法体外分离培养骨髓瘤骨髓基质细胞和人脐血源基质细胞,与骨髓瘤KM3细胞共培养。采用扫描电镜观察共培养后基质细胞和KM3细胞的位相关系;应用CCK-8、PI染色流式细胞仪和AnnexinV/PI双标法检测不同培养条件下KM3细胞的增殖、细胞周期和凋亡率。结果人脐血基质细胞比骨髓瘤患者骨髓基质细胞有更强的抑制KM细胞增殖的作用;共培养后骨髓瘤患者骨髓基质细胞使KM3细胞阻滞于G0/G1期[(59.70±1.28)%],人脐血基质细胞使S期的KM3细胞比例增高[(46.07±2.46)%];KM3/MM-BMSCs组KM3细胞凋亡率为(3.26±0.12)%明显低于KM3/hUCBDSCs组KM3细胞凋亡率(4.76±0.12)%(P<0.01);KM3/MM-BMSCs组KM3细胞BaxmRNA的表达较KM3/hUCBDSCs组明显下降(P<0.01);KM3/hUCBDSCs组KM3细胞Bcl-2mRNA的表达较KM3细胞悬浮培养组下降(P<0.05),KM3/MM-BMSCs组KM3细胞Bcl-2mRNA的表达则明显升高,为悬浮培养组的1.982倍(P<0.01)。结论骨髓瘤基质细胞微环境抑制骨髓瘤细胞凋亡和死亡;而人脐血源基质细胞有更强的抑制骨髓瘤细胞增殖的作用,并促进其凋亡。
Objective To observe the effects of bone marrow stromal cells (BMSCs) on the biological characteristics of myeloma KM3 cells in human umbilical cord blood stromal cells and myeloma patients. Methods Myeloma bone marrow stromal cells and human umbilical cord blood-derived stromal cells were isolated and cultured in vitro and co-cultured with myeloma KM3 cells. The relationship between the stromal cells and KM3 cells was observed by scanning electron microscopy. The proliferation, cell cycle and apoptosis of KM3 cells were detected by CCK-8, PI staining and Annexin V / PI double labeling. . Results Human umbilical cord blood stromal cells inhibited the proliferation of KM cells more than those of myeloma patients. KM3 cells were arrested in G0 / G1 phase after co-cultured with myeloma [(59.70 ± 1.28)%] (46.07 ± 2.46)%]. The apoptosis rate of KM3 cells in KM3 / MM-BMSCs group (3.26 ± 0.12)% was significantly lower than that in KM3 / hUCBDSCs group KM3 (4.76 ± 0.12)% (P <0.01). The expression of Bax mRNA of KM3 cells in KM3 / MM-BMSCs group was significantly lower than that of KM3 / hUCBDSCs group (P <0.01). The expression of Bcl-2 mRNA in KM3 / hUCBDSCs group was significantly higher than that in KM3 / The expression of Bcl-2 mRNA in KM3 cells was significantly higher than that in KM3 cells (P <0.05), which was 1.982 times higher than that in KM3 cells (P <0.01). Conclusion Myeloma stromal microenvironment inhibits the apoptosis and death of myeloma cells. Human umbilical cord blood derived stromal cells inhibit the proliferation of myeloma cells and promote their apoptosis.