尿多酸肽联合亚砷酸对K562细胞凋亡及细胞增殖的影响

Effect of CDA-2 Combined with Arsenous Acid on the Apoptosis and Proliferation of K562 Cells

【目的】观察尿多酸肽（cDA-2）、亚砷酸、CDA_2与亚砷酸联合体外诱导慢性髓细胞白血病细胞（K562）株凋亡、抑制增殖的作用；探讨CDA-2与亚砷酸联合对诱导K562细胞凋亡及抑制增殖的协同作用。【方法】不同浓度的CDA-2、亚砷酸及二者联合处理K562细胞株，通过MTT比色法检测K562细胞细胞生长率，计算IC50值；应用倒置相差显微镜、A0荧光染色观察细胞凋亡及形态学变化；应用流式细胞术检测细胞凋亡。【结果】①MTT结果显示，在一定的范围内，K562细胞的生长率存在剂量及时间依赖性降低，CDA-2作用K562细胞24h、48h、72h的IC50值分别为75mg／L、59mg／L和37mg／L，亚砷酸作用K562细胞24h、48h、72h的IC50值分别为11．55μmol／L、6．14gmol／L及5．18μmol／L；②荧光染色结果显示，与未加药的对照组相比，二者单药均可促进细胞凋亡，二者联合后凋亡作用更加显著；③流式细胞术显示，cDA-262．5mg／L处理K562细胞24h早期凋亡细胞为（1．77±0．49）％，亚砷酸4umol／L处理K562细胞24h早期凋亡细胞为（1．33±0．58）％，而相同浓度的二者联合用药作用于K562细胞24h早期凋亡细胞为（11．03±0．7）％（P〈0．05），按照预计效应公式得出，两药联合存在协同作用，且协同作用于24h，两药最低浓度联合时已开始，随着两药浓度的增加，协同作用逐渐增大。【结论】CDA-2、亚砷酸单药及二者联合均可抑制K562细胞株增殖并促进凋亡，二者具有协同作用。

[Objective]To observe the role of uric polyaeid peptide（CDA-2）, arsenous acid and their com- bination in inducing apoptosis and inhibiting proliferation of chronic myelogenous leukemia cells （K562） in vitro, and explore the synergetic effect of two drugs in inducing K562 cell apoptosis and proliferation. [Meth- ods] Different concentrations of CDA-2, arsenous acid and their combination were used to treat K562 cells. K562 cell growth rate was detected by using MTT colorimetric assay, and IC50 was calculated. Inverted phase contrast microscope and AO fluorescent staining were used to observe cell apoptosis and morphological chan- ges. Cell apoptosis was measured by flow cytometry. [Results]MTT results showed that the growth rate of K562 cells decreased in a dose- and time-dependent manner in a certain range. ICS0 of CDA-2-treated K562 cells at 24h, 48h and 72h were 75mg/L, 59mg/L and 37mg/L, respectively. IC50 of arsenous acid-treated K562 cells at 24h, 48h and 72h were 11.55μmol/L, 6.14μmol/L and 5.18μmol/L, respectively. Fluorescent staining results showed that both CDA-2 and arsenous acid could promote cell apoptosis, and CDA-2 combined with arsenous acid could significantly increase cell apoptosis in comparison with control group without drug treatment. Flow cytometry showed that early cell apoptosis rate of K562 cells treated with 62.5mg/L CDA-2 at 24h was （1. 77±0. 49）%, while that of K562 cells treated with 4μmol/L arsenous acid was （1. 33±0.58） %. The early cell apoptosis rate of K562 cells treated with CDA-2 combined with arsenous acid at 24h was （11.03±0.70） %（ P 〈0.05）. According to the expected effect formula, the combined drugs had syner-gistic effect. The synergistic effect began at 24h while the two drugs were minimum concentration. With the increasing of the concentration of two drugs, the synergistic effect increased gradually. [Conclusion]CDA-2, arsenous acid or their combination can inhibit cell proliferation and promote cell apoptosis of K562 cells. The combination of two drugs has synergistic effect.