巴豆生物碱上调TRAIL配体、caspase-8的表达诱导HeLa细胞凋亡的体外研究

Apoptosis Induced by Croton Alkaloid (CA) on Cervical Cancer HeLa Cells through Up-regulation of TRAIL and Caspase-8 in vitro

目的研究巴豆生物碱对人宫颈HeLa细胞诱导凋亡作用,并探讨其作用机制。方法 MTT法观察巴豆生物碱对HeLa细胞的抑制率;流式细胞仪检测HeLa细胞TRAIL及其受体DR5的表达及其巴豆生物碱对HeLa细胞TRAIL配体表达的影响;Caspase-8活性检测;RT-PCR检测TRAIL配体及Caspase-8、Caspase-3 mRNA的表达。结果巴豆生物碱对HeLa细胞生长有抑制作用,且呈剂量依赖关系。流式细胞仪检测HeLa细胞TRAIL及其受体DR5的表达阳性,巴豆生物碱处理HeLa细胞后发现TRAIL配体的表达明显上调,Caspase-8活性明显增加,TRAIL配体、Caspase-3及Caspase-8 mRNA高表达。结论巴豆生物碱对人宫颈癌HeLa细胞有增殖抑制和诱导细胞凋亡的作用,其诱导机制可能与上调TRAIL配体和Caspase-8mRNA的表达有关。

OBJECTIVE To investigate the effect of the Croton Alkaloid on proliferation and apoptosis of HeLa human cervical cancer cells in vitro and explore its probable molecular mechanisms.METHODS Inhibition of the cell proliferation was measured by MTT assay.The expressions of TRAIL、death receptors 5 and TRAIL ligand before and after CA treatment were detected by FCM.The Caspase-8 activity was detected by Caspase-8 Activity Assay Kit and the expressions of TRAIL ligand and caspase-8,caspase-3 mRNA was tested by RT-PCR.RESULTS CA inhibited the proliferation of HeLa cells in a dose-dependent manner.Positive expressions of TRAIL and death receptor 5 were detected by FCM.The expressions of TRAIL ligand、 Caspase-8 activity was up regulated obviously and the level of TRAIL、 Caspase-3、Caspase-8 mRNA was increased after CA treatment.CONCLUSION CA displays a proliferation inhibitory apoptosis-inducingeffect in HeLa cells.The molecular mechanisms of apoptosis HeLa induced by CA has possibly related to up regulating the expresstion of TRAIL ligand and Caspase-8 activity