β-细辛醚对肠癌细胞HT-29生物行为学的影响及机制

Effects of β-asarone on biological behavior and its mechanism of Conlon cancer cells HT-29 in vitro

目的观察β-细辛醚对肠癌细胞株体外增殖抑制作用及其机制。方法①MTT法观察β-细辛醚对多种肿瘤细胞株的增殖的影响。②Transwell小室法,观察β-细辛醚对肠癌细胞株HT-29侵袭的影响;③粘附实验观察β-细辛醚对HT-29粘附能力的影响;④AnnexinV/PI双染法研究β-细辛醚对肠癌细胞株HT-29凋亡的诱导作用;⑤流式细胞仪检测β-细辛醚对肠癌细胞株HT-29细胞周期的影响;⑥JC-1荧光探针法检测线粒体膜电位水平的改变;⑦流式细胞仪检测β-细辛醚对肠癌细胞株HT-29活性氧水平的改变。结果①β-细辛醚对多种肿瘤细胞株有不同程度的抑制作用(P<0.05)。②β-细辛醚作用肠癌细胞HT-29后,细胞的侵袭和粘附能力明显降低(P<0.05)。③β-细辛醚作用肠癌细胞株HT-29后出现明显的早期凋亡及细胞周期明显改变。④β-细辛醚作用肠癌细胞株HT-29后线粒体膜电位降低及内活性氧水平升高。结论①β-细辛醚对肠癌细胞有明显的增殖抑制作用,能降低肠癌细胞HT-29的粘附和侵袭能力。②β-细辛醚可诱导肠癌细胞凋亡,改变细胞周期,其机制可能与线粒体凋亡途径有关。

Objective This study was designed to explore whether β asarone can inhibit the proliferation of human conlon cells HT -29 in vitro and its mechanism. Methods （1）M3T method was used to observe the effects of β-asarone on human conlon cells;（2）Transwell invasion assay was carried out to quantify the invasion capability after treatment; （3）Adhesion capability of HT -29 cells was evaluated by MTT method;（4）Flow eytometry was applied to verify the early apoptosis and the cell cycles induced by different concentrations in human conlon cells HT- 29 ;（5）Fluorescent probe JC - 1 was used to observe the changes in mito- chondrial membrane potential ; （6）JC - 1 fluorescent probe method to detect changes in the level of mitochondria membrane poten- tial ;（7）Flow cytometry detect changes in the level of reactive oxygen species. Results （1）β-asarone inhibited the proliferation of various human conlon cancer cells among which HT -29 was the most sensitive. （2）The adhesion and invasion capability of HT - 29 cells decreased after β - asarone treatment （ P 〈 0.05 ）. （3）β - asarone can induce the cells apoptosis and change cell cycle. （4） - asarone can decrease the cell mitoehondrial membrane potential and active oxygen levels. Conclusion β - asarone can signifi- cantly change the HT - 29 biological behavior, including adhesive and invasive ability. β - asarone can induce apoptosis through the mitochondrial apoptotic pathway and change the cell cycle.