Objective: To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism.
Methods: MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry.
Results: The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G2/M phase increased (r=0.88), and cells were blocked in G2/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS.
Conclusion: DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.
题目: 黄花败酱茎秆二氯甲烷萃取相对人白血病细胞K562增殖及分化的影响.
目的: 探讨黄花败酱茎秆乙醇提取物的二氯甲烷萃取相（DPSS）对K562细胞增殖抑制和诱导分化作用及其相关作用机制.
方法: 不同浓度DPSS（0、25、50、100和200 μg/ml）分别作用于细胞24、48和72 h后，采用MTT法检测K562细胞增殖情况；应用流式细胞术检测不同浓度DPSS对K562细胞凋亡、周期阻滞的影响；应用瑞氏-吉姆萨染色观察DPSS作用后K562细胞形态学的变化并应用流式细胞术验证分化相关抗原CD33、CD11b的表达.
结果: 与对照组比较，加药组各浓度DPSS均对K562细胞的增殖有明显抑制作用，并且存在时间剂量依赖性（r=-0.96）；细胞周期分析结果表明，随着DPSS浓度升高，G2/M期细胞增多（r=0.88），细胞被阻滞在G2/M期；流式细胞术结果显示，当200 μg/ml DPSS作用48 hK562细胞凋亡率最高；瑞氏-吉姆萨染色结果显示，DPSS作用后K562细胞出现胞体增大，包浆量增多，核浆比减小，核染色质粗糙等分化表现；细胞分化抗原检测发现，DPSS作用后CD33、CD11b呈阳性表达.
结论: DPSS可以通过周期阻滞诱导K562细胞凋亡、抑制其增殖，并且能够诱导K562细胞向单核细胞分化，具有潜在抗白血病细胞的作用.
Keywords: K562 cell line; Patrinia scabiosaefolia Fisch. Stem; cell apoptosis; dichloromethane extraction phase; induced differentiation.