小白菊内酯诱导人神经母细胞瘤SK—N—SH细胞凋亡的研究

The effects of parthenolide on proliferation and apoptosis of human neuroblastoma cell line SK-N-SH

目的 探讨小白菊内酯对人神经母细胞瘤细胞系SK～N—SH细胞凋亡的影响及机制。方法MTT法检测不同浓度小白菊内酯对SK—N—SH细胞增殖抑制的影响，同时计算IC50，根据IC50选取实验组小白菊内酯浓度（10、15μmol／L）；流式细胞仪分析对照组及实验组肿瘤细胞凋亡率；EMSA检测NF-kB的活性；WesternBlot法检测细胞凋亡相关蛋白表达。结果小白菊内酯抑制神经母细胞瘤SK—N—SH细胞生长，并呈浓度依赖关系。用10、15μmol／L小白菊内酯处理细胞48h后凋亡率分别为（35．63±0．98）％，（53．32±1．08）％，与对照组（6．42±1．26）％比较，差异显著（P〈0．01）。EMSA结果表明，小白菊内酯处理组细胞NF_KB活性降低。WesternBlot结果显示，小白菊内酯使细胞中Bax、Caspase-3蛋白表达增强，Bel-2蛋白表达降低。结论小白菊内酯促进神经母细胞瘤SK-N-SH细胞凋亡，可能与抑制NF-xB活性，下调Bcl-2蛋白表达，上调Bax、Caspase-3蛋白表达有关。

Objective To explore the effects of parthenolide on proliferation and apoptosis of human neuroblastoma cell line SK-N-SH. Methods SK-N-SH, a human neuroblastoma cell line, was treated with different concentrations of parthenolide for 48h, the cellular proliferative activities were examined by MTT assay. The parthenolide concentration of experimental group （10 mol/L, 15 mol/L） was determined by I（350. Apoptosis rate of SK-N-SH cells was analyzed by flow cytometry （FCM）. The activity of NF-kB of SK-N-SH cells after parthenolide treatment was assayed by EMSA. Western Blot was used to detect protein expression of Bcl-2, Bax, Caspase-3. Results The proliferative activities of SK-N-SH treated with parthenolide were obviously suppressed, the proliferative rate decreased in a dose-dependent manner. Treated with 10 mol/L and 15 mol/L parthenolide after 48h, the apoptosis rate was 35.63 ±0. 98% and 53.32 ± 1.08%, respectively. Compared with the control group （6. 42 ± 1,26 %）, the differences were significant （P〈0. 01）. EMSA results showed that the activity of NF-kB in SK-N-SH cells after parthenolide treatment for 48h was decreased. Western Blot showed that parthenolide increased protein levels of Bax and Caspase-3 and decreased protein level of Bcl-2 in SK-N- SH cells. Conclusions Parthenolide can suppress the proliferative activity of neuroblastoma cell line SK-N-SH and induce cell apoptosis. The anti-tumor mechanism of parthenolide may be related to the inhibited NF-kB, reduced Bcl-2 protein expression and increased Bax, Caspase-3 proteins.