与白血病骨髓间充质干细胞共培养后K562细胞增殖与凋亡的变化

Changes of proliferation and apoptosis of K562 cells after cultivation with leukemia bone marrow mesenchymal stem cells

目的比较K562细胞与人白血病骨髓间充质干细胞（LMSC）共培养前后增殖和凋亡的变化。方法采用细胞计数试剂盒-8（CCK-8）分析法检测SCG和CCG组K562细胞光密度（OD）值,比较不同培养条件下K562细胞的增殖情况。采用流式细胞仪检测与LMSC共培养后K562细胞周期变化,Annexin V/聚酰亚胺（PI）荧光标记法检测血清饥饿后与LMSC共培养后K562细胞凋亡的变化。结果与LMSC共培养后,K562细胞的增殖受到明显抑制,CCG组OD值明显低于SCG组。流式细胞仪检测细胞周期发现,与LMSC共培养后,G0~G1期细胞比例明显增高,S期细胞比例明显下降,且G2~M期细胞比例也呈上升趋势;CCG＋0%FBS组K562细胞凋亡较SCG＋10%FBS组明显增多,较SCG＋0%FBS明显降低,LMSC有抵抗白血病细胞凋亡的作用。结论 LMSC通过阻滞G0~G1期K562细胞周期而发挥抑制增殖的作用,并能抑制血清饥饿诱导的K562细胞凋亡。

Objective To compare the changes of proliferation and apoptosis of K562 cells after cultivation with human leukemia bone marrow mesenchymal stem cells（LMSC）.Methods Cell counting kit-8（CCK-8） analytic approach was adopted to detect the optical density（OD） value of K562 cells in SCG and CCG groups,and the conditions of K562 cell proliferation under different cultured conditions were compared.Flow cytometer（FCM） was used to detect K562 cell cycle changes after cultivation with LMSC,Annexin V/polyimide（PI） fluorescence labeling method to detect K562 cell apoptosis changes after cultivation with LMSC and serum starvation.Results After cultivation with LMSC,the proliferation of K562 cells was markedly inhibited,and OD value in CCG group was conspicuously lower than that in SCG group.FCM detection to cell cycles demonstrated that after cultivation with LMSC,the proportion of cells at G0~G1 and G2~M phases notably increased,whereas that at S phase obviously decreased.The increase of K562 cell apoptosis in CCG＋0%FBS group was more significant than that in SCG＋10%FBS group,while its decrease was more remarkable compared with that in SCG＋0%FBS group.LMSC had the function of resisting leukemia cell apoptosis.Conclusion LMSC exerts the effect of inhibiting the proliferation by blocking K562 cell cycles at G0~G1 phase,and can inhibit K562 cell apoptosis induced by serum starvation.