摘要
目的探讨丙戊酸(VPA)对神经母细胞瘤细胞自噬性死亡的调控。方法用含不同浓度的VPA培养基培养SH-SY5Y细胞,应用MTT法检测细胞的存活率,乳酸脱氢酶(LDH)检测细胞损伤,单丹磺酰尸胺(MDC)染色评价细胞质内酸性自噬泡的形成,Western blotting法检测巨自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ的比值及Beclin-1的表达。结果与对照组比较,VPA孵育48 h后,SH-SY5Y细胞存活率明显降低,且具有剂量依赖性(P<0.05);SH-SY5Y细胞培养基中LDH明显增高,且具有剂量依赖性(P<0.05);VPA诱导细胞内LC3-Ⅱ/LC3-Ⅰ的比值及Beclin-1的表达水平明显上调(P<0.05)。结论 VPA诱导神经母细胞瘤细胞细胞自噬性死亡,为神经母细胞瘤的治疗提供新的手段。
Objective To investigate the effect and mechanism of valproic acid（ VPA）on autophagic cell death in neuroblastoma SH-SY5Y cells. Methods We used different concentration of VPA to treat SH-SY5Y cells. MTT was used to detect the viability of SH-SY5Y cells,layered double hydroxides（LDH）was used to evaluate the damage of cells,monodansylcadaverine（ MDC）staining was used to mark autophagic vacuoles. The autophagy-related protein expressions of LC3-Ⅱ/LC3-Ⅰ and Beclin-1 were detected by Western blotting analysis. Results After treatment of VPA for 48 hours,the viability of SH-SY5Y cells were significantly decreased in dose-dependent manner（P〈0. 05）;and VPA increased LDH level in culture media in dose- dependent manner（P〈0. 05）. Compared with the control group,the levels of LC3-Ⅱ/LC3-Ⅰand Beclin-1 were significantly increased in the VPA treatment groups（P〈0. 05）. Conclusion VPA induces the autophagic cell death of neuroblastoma SH-SY5Y cells,VPA thus may be a new target for treatment of neuroblastoma.
出处
《肿瘤基础与临床》
2015年第2期93-95,共3页
journal of basic and clinical oncology
基金
国家自然科学基金资助项目(编号:81241046)
