Medical College of Chinese People's Armed Police Forces, Tianjin 300162, China.
Eur J Pharmacol. 2010 Feb 10;627(1-3):69-74. doi: 10.1016/j.ejphar.2009.10.056. Epub 2009 Oct 30.
Our group synthesized a new potent anti-tumor podophyllotoxin derivative, YB-1EPN. In our study, we found that it was more potent than etoposide (VP-16). Interestingly, we found that the KBV200 cell line and K562/A02 cell line were rendered resistant towards VP-16 but not towards YB-1EPN. In vitro, both the cytotoxicity of YB-1EPN and its ability to inhibit KBV200 and K562/A02 cells were determined by MTT assay and growth curve. The IC(50) value of YB-1EPN on KBV200 cell was (2.52+/-0.28)microM in contrast to VP-16 (10.1+/-0.220)microM. And YB-1EPN showed a dose-dependent and broad-spectrum antiproliferative activity. Inducing apoptosis by YB-1EPN in KBV200 was assessed by various morphological and biochemical characteristics, including cell shrinkage, chromatin condensation, membrane blebbing, formation of apoptotic bodies, and DNA ladder formation. Rates of apoptosis and cell cycle were also checked through flow cytometry. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect mdr-1,p53,bcl-2,and bax gene expression. Western-blot assay was used to assess P-glycoprotein (P-gp) expression. We found that YB-1EPN could down-regulate expression level of the mdr-1, bcl-2, and up-regulate expression level of p53, and bax mRNA, as compared to the control. These results suggest that YB-1EPN has the potentiality to overcome P-glycoprotein-mediated multidrug resistance in the KBV200 cell line.
我们小组合成了一种新型有效的抗肿瘤鬼臼毒素衍生物 YB-1EPN。在我们的研究中，发现它比依托泊苷（VP-16）更有效。有趣的是，我们发现 KBV200 细胞系和 K562/A02 细胞系对 VP-16 产生耐药性，但对 YB-1EPN 没有耐药性。在体外，通过 MTT 检测和生长曲线来确定 YB-1EPN 的细胞毒性及其抑制 KBV200 和 K562/A02 细胞的能力。YB-1EPN 对 KBV200 细胞的 IC50 值为（2.52+/-0.28）μM，而 VP-16 为（10.1+/-0.220）μM。YB-1EPN 表现出剂量依赖性和广谱的抗增殖活性。通过各种形态学和生化特征评估 YB-1EPN 在 KBV200 中诱导凋亡的情况，包括细胞收缩、染色质浓缩、细胞膜起泡、凋亡小体形成和 DNA 梯形成。通过流式细胞术检查凋亡和细胞周期的比率。逆转录-聚合酶链反应（RT-PCR）用于检测 mdr-1、p53、bcl-2 和 bax 基因表达。Western-blot 检测用于评估 P-糖蛋白（P-gp）表达。我们发现，与对照组相比，YB-1EPN 可以下调 mdr-1、bcl-2 的表达水平，并上调 p53 和 bax mRNA 的表达水平。这些结果表明，YB-1EPN 有可能克服 KBV200 细胞系中 P-糖蛋白介导的多药耐药性。
