体外观察中华眼镜蛇毒组分对KG1a细胞增殖抑制作用及对其细胞周期的影响

Naja Naja Actra Venom Component Inhibits Proliferation and Affects the Cell Cycle in vitro of KGla Cells

目的:探讨中华眼镜蛇毒组分(NaiaNaiaActravenomcomponent,NNAVC)对KG1a细胞的增殖抑制作用及对其细胞周期的影响。方法:以人急性髓系白血病细胞株KG1a细胞为研究对象进行实验。应用不同剂量的中华眼镜蛇毒组分作用于KG1a细胞后,采用MTT法测定其对KG1a细胞的生长抑制作用,倒置显微镜镜下观察药物作用前后细胞形态学变化,流式细胞仪检测中华眼镜蛇毒组分对KG1a细胞周期的影响。结果:中华眼镜蛇毒组分可以明显的抑制KG1a细胞的增殖。在0.7μg/mL至1.2μg/mL的浓度范围内,中华眼镜蛇毒组分作用具有时间依赖性及剂量依赖性。显微镜镜下观察KG1a细胞经中华眼镜蛇毒组分作用6h后状态开始发生变化,随着药物作用时间的延长,细胞逐渐出现体积变小、变形、胞内空泡、细胞固缩等形态学变化。流式细胞仪检测证实中华眼镜蛇毒组分作用后,能够将KG1a细胞阻滞于细胞周期的G_0/G_1期,而处于G_2/M期的细胞含量减少。结论:中华眼镜蛇毒组分对KG1a细胞的增殖具有明显抑制作用,调节细胞周期停滞于G_0/G_1期,减少G_2/M期的细胞含量,提示影响细胞周期进程可能是中华眼镜蛇毒组分对KG1a细胞的发挥增殖抑制作用的机制之一。

Objective： To investigate the effect of Naja Naja Actra Venom Component （NNAVC） on the proliferation and cell cycle of KG1a cells. Methods： The human acute myeloid leukemia （AML） cell KGla was chosen as the experimental subject. After exposure to different concentrations of NNAVC, MTT assay was used to detect the inhibitory action of NNAVC on KGla cells. Morphological changes of KGla cells were observed by inverted microscope before and after administration of NNAVC. Cell cycles were measured via flow cytometry. Results： NNAVC obviously inhibited the proliferation of KGla cells. The results suggest NNAVC inhibited the KGla ceils in a time- and dose-dependent manner within the range of （0.7-1.2） μg/mL. Smaller volume, deformation, vacuole within the cell, cell pyknosis and other morphological changes were gradually observed by inverted microscope after 6 hours. After treatment with NNAVC, flow cytometry analysis showed KGla cells were arrested in G0/G1 stage and significantly decreased in G2/M stage. Conclusion： NNAVC can obviously inhibit the proliferation of KGla cells. KG1a cells were arrested in G0/G1 stage and significantly decreased in G2/M stage. This data suggests the change of cell cycle progression may be one of the underlying mechanisms of NNAVC inhibiting the growth of KGla cells.