ZM-66, a New Podophyllotoxin Derivative Inhibits Proliferation and Induces Apoptosis in K562/ADM Cells

Ling Li; Hong-jie Li; Jian-sheng zhi; Hong Chen; Wen-li Xie

摘要

Objective To investigate the anti-tumor effect of ZM-66 on multidrug-resistant leukemic cell line K562/ADM. Methods The K562/ADM cells were treated with varying concentrations (0, 1, 2, 4×10-3 mmol/L) of ZM-66 or etoposide for 24 hours. The proliferation was detected by Sulforhodamine B Sodium Salt (SRB) assay and apoptosis was detected by flow cytometry analysis and fluorescent staining. In addition, the expression levels of p53 and bax genes in K562/ADM cells were detected by RT-PCR analysis. The level of P-glycoprotein (P-gp), P53 and Bax protein in K562/ADM cells were detected by Western blot assay. Results SRB assay demonstrated that etoposide had little inhibitory effect on K562/ADM cells, whereas ZM-66 (1, 2, 4×10-3 mmol/L) had significantly inhibitory effect on K562/ADM cells (all P<0.01). The acridine orange/propidium iodide dual staining showed that there were typical condensation of chromatin and nuclear fragmentation nuclei with red color in ZM-66 treated cells. Flow cytometric analysis showed that there was a significantly increase of apoptotic cells in K562/ADM cells after treated with ZM-66. RT-PCR showed that the p53 and bax mRNA expression levels in K562/ADM cells treated with ZM-66 at 1, 2, 4×10-3 mmol/L were higher than those in the cell without treatment. Western blot showed that the P53 and Bax protein expression levels in K562/ADM cells treated with ZM-66 at 2, 4×10-3 mmol/L were higher than those in the cell without treatment. But the P-gp protein expression level in K562/ADM cells treated with ZM-66 at 2, 4×10-3 mmol/L was gradually lower than those in the cell without treatment. Conclusion ZM-66 is able to induce cell death by apoptosis in vitro, as a result of the reverse of the apoptosis resistance in drug-resistant K562/ADM cells by modulating expression of key factors associated with apoptosis induction.

机译：目的研究ZM-66对多药耐药白血病细胞株K562 / ADM的抗肿瘤作用。方法用不同浓度（0、1、2、4×10-3 mmol / L）的ZM-66或依托泊苷处理K562 / ADM细胞24小时。磺胺丁丹B钠盐（SRB）检测增殖，流式细胞仪分析和荧光染色检测凋亡。另外，通过RT-PCR分析检测K562 / ADM细胞中p53和bax基因的表达水平。用Western blot法检测K562 / ADM细胞中P-糖蛋白（P-gp），P53和Bax蛋白的水平。结果SRB分析表明，依托泊苷对K562 / ADM细胞几乎没有抑制作用，而ZM-66（1、2、4×10-3 mmol / L）对K562 / ADM细胞具有明显的抑制作用（所有P < 0.01）。 orange啶橙/碘化丙啶双重染色表明，在ZM-66处理的细胞中，染色质和核碎裂核具有典型的缩合，呈红色。流式细胞仪分析显示，ZM-66处理后，K562 / ADM细胞中凋亡细胞明显增加。 RT-PCR显示，分别用1、2、4×10-3 mmol / L的ZM-66处理的K562 / ADM细胞中的p53和bax mRNA表达水平高于未处理的细胞。 Western blot检测显示，在ZM-66处理2、4×10-3 mmol / L的K562 / ADM细胞中，P53和Bax蛋白表达水平高于未处理的细胞。但是在ZM-66处理2、4×10-3 mmol / L的K562 / ADM细胞中，P-gp蛋白表达水平逐渐低于未处理的细胞。结论结论ZM-66能够通过体外凋亡诱导细胞死亡，这是由于通过调节与凋亡诱导相关的关键因子的表达逆转了耐药性K562 / ADM细胞的凋亡抗性。

ZM-66, a New Podophyllotoxin Derivative Inhibits Proliferation and Induces Apoptosis in K562/ADM Cells

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