熊猫豆提取物抑制肿瘤活性及诱导结肠癌细胞Caco-2凋亡机制研究

Anti-Cancer Activity of Panda Bean Extract and Apoptosis-Inducing Mechanism Against Caco-2 Cells

为了探究熊猫豆的抑制肿瘤活性及其诱导Caco-2凋亡的机制，采用80％丙酮浸提法制备熊猫豆粗提物，通过MrIYr法测定其对人结肠癌细胞Caco-2增殖的抑制效果，琼脂糖凝胶电泳、流式细胞术、酶联免疫分析等技术与方法检测细胞周期、DNA及凋亡相关蛋白相对表达量的变化。结果发现，与空白对照组相比，0．1～1．0mg／mL熊猫豆丙酮提取物对Caco-2细胞增殖有明显的抑制作用，且呈现剂量效应关系，IC50为0．72mg／mL。0．4、0．6、0．8mg／mL熊猫豆丙酮提取物处理72h后的Caco-2细胞，可以观察到典型的凋亡细胞的梯状DNA条带，细胞周期G1期延长，CytC、caspase-9、caspase-3蛋白的相对表达量均有显著上升。熊猫豆丙酮能够抑制结肠癌细胞Caco-2增殖，这种抑制作用是通过阻滞细胞周期和诱导线粒体通路介导的细胞凋亡实现的。

In order to research the anti - cancer activity of panda bean extract and its mechanism of apoptosis - inducing against Caco -2 cells, active compounds existed in panda bean have been extracted by 80% acetone aqueous solution. The inhibitory effect of panda bean extract against colon cancer cell Caco - 2 with various concentrations was determined by MTT method. PI staining has been utilized to detect by cell cycle. Agarose gel electrophoresis has been utilized to detect cell apoptosis, and apoptosis -inducing mechanism was detected by ELISA. The results showed that compared with control group, 0.1 - 1.0 mg/mL panda bean extracts expressed a significant inhibitory effect against Caco - 2 with a dose - effect relationship and ICs0 was 0.72 mg/mL. After treated by panda bean extracts （0.4, 0.6 and 0.8 mg/mL） for 72 h, typical apoptotic cell DNA ladder could be observed. Flow cytometry analysis showed that Caco - 2 cell cycle could be arrested at G1 phase and sub - G1, which symbolized that the apoptosis cell was appeared after being exposed to panda bean extract. In addition, CytC, caspase -9 and caspase -3 protein levels were significantly increased. The results suggested that panda bean extract could induce apoptosis via mitoehondrial pathway.