β-榄香烯诱导神经母细胞瘤细胞凋亡及其机制

β-elemene induces neuroblastoma cell apoptosis

目的探讨β-榄香烯对儿童神经母细胞瘤生长的抑制作用及其可能的机制。方法不同质量浓度β-榄香烯体外作用于神经母细胞瘤细胞株SH-SY5Y、IMR-32、SK-N-DZ及胚肾细胞株293T 24 h,用CCK-8法检测细胞存活率,流式细胞术检测SH-SY5Y及SK-N-DZ细胞的凋亡率,Hoechst 33342染色检测凋亡小体,流式细胞术检测细胞周期和Brd U掺入率,Western blot检测caspase-3、caspase-8及caspase-9蛋白表达。结果β-榄香烯对3株神经母细胞瘤细胞的生长均有抑制作用,且呈剂量依赖性,而对正常细胞抑制作用轻微;β-榄香烯作用于SH-SY5Y及SK-N-DZ细胞24 h后,与对照组相比,药物组细胞早期凋亡率上升(P<0.05),凋亡小体明显增加(P<0.05);细胞各周期比例无明显差异(P>0 05);Brd U掺入率无明显差异(P>0.05);与对照组相比,药物组caspase-3及caspase-9蛋白表达明显增多(P<0.05),而caspase-8无明显差异(P>0.05)。结论β-榄香烯对神经母细胞瘤细胞的生长具有抑制作用,其作用机制是通过激活内源性凋亡通路诱导细胞凋亡,而与细胞周期及细胞增殖无明显关系。

This study was performed to investigate the therapeutic effect of β-elemene on the neuroblastoma in vitro. Neuroblastoma cell lines including SH-SY5 Y, IMR-32, SK-N-DZ and embryo kidney cell line 293 T weretreated with β-elemene in different dose and duration, then cell viability was measured by cell counting kit-8（CCK8） assay. Cell apoptosis was measured by 7 AAD/annexin V staining and Hoechst 33342 staining. Data showedthat the cell viability of neuroblastoma were decreased when treated with β-elemene. Compared with the PBStreated control group, the rate of early apoptosis of SH-SY5 Y and SK-N-DZ cells was increased, and apoptoticbodies were increased obviously, however, the proportion of cell cycles and incorporation of Brd U in theneuroblastoma were unaltered after treatment with β-elemene for 24 h. Western blotting showed that the expressionof caspase-3 and caspase-9 were increased significantly in neuroblastoma after treatment with β-elemene in adose-dependent manner, but there was no significant difference in the expression of caspase-8. Together, these datasuggest that β-elemene inhibits the growth of neuroblastoma due to the induction of mitochondrialpathway-associated apoptosis, but not arresting cell cycle and proliferation.