摘要
目的观察β-榄香烯对人胰腺癌Panc-1细胞凋亡的影响,探讨其作用机制。方法β-榄香烯浓度设置为10、20、40、80、160μg/m L,作用于体外培养的Panc-1细胞24、48、72 h。台盼蓝拒染法检测Panc-1细胞抑制率;TUNEL法检测Panc-1细胞凋亡;Hoechst33258荧光染色观察Panc-1细胞核变化;ELISA检测Panc-1细胞Caspase-3、8、9活性;Western blot检测Panc-1细胞Fas、Fas L、细胞色素C(Cyt c)、凋亡诱导因子(AIF)蛋白表达。结果与对照组比较,β-榄香烯作用Panc-1细胞24、48、72 h,Panc-1细胞抑制率明显增加(P<0.05,P<0.01),细胞凋亡率明显增加(P<0.01,P<0.001),并呈时间/浓度依赖性;β-榄香烯作用Panc-1细胞72 h,Panc-1细胞核可见明显碎裂,染色质浓缩,呈强蓝色荧光,形成凋亡小体;β-榄香烯作用Panc-1细胞48 h,Caspase-3、8、9活性明显增加(P<0.05,P<0.01),Fas、Fas L、Cyt c及AIF蛋白表达明显增强(P<0.05,P<0.01,P<0.001)。结论β-榄香烯能够抑制Panc-1细胞增殖、诱导细胞凋亡,其可能激活细胞内死亡受体途径及线粒体凋亡途径发挥抗肿瘤作用。
Objective To investigate the effects of β-Elemene on the apoptosis of human pancreatic cancer Panc-1 cells; To discuss its mechanism of action. Methods β-Elemene（10, 20, 40, 80, 160 μg/m L） were incubated to the Panc-1 cells in vitro cultured for 24 h, 48 h and 72 h, and trypan blue refusal method was used to detect cell inhibition rate; Apoptosis rate was measured by TUNEL; Hoechst33258 fluorescent staining was used to observe the changes of the nucleus. The activity of Caspase-3, 8 and 9 were detected by ELISA. Western blot was used to detect the expressions of Fas, Fas L and Cyt c and AIF. Results The activity of Panc-1 cells was obviously inhibited time/concentration dependent inhibition（P〈0.05, P〈0.01）, and the apoptosis rate increased after incubated with β-Elemene（P〈0.01, P0.001） after incubated with β-Elemene for 24 h, 48 h and 72 h; After giving β-Elemene 72 h, Panc-1 cells nucleus were broken obviously, and chromatin condensed and showed strong blue fluorescence, along with of apoptotic bodies; After incubated with β-Elemene for 48 h, Caspase-3, 8 and 9 activity significantly increased（P〈0.05, P〈0.01）; protein expressions of Fas, Fas L, Cyt c and AIF were significantly enhanced（P〈0.05, P〈0.01, P〈0.001）. Conclusion β-Elemene can inhibit Panc-1 cell proliferation, induce apoptosis, and the mechanism may be related to activating cell death receptor pathway and mitochondrial apoptosis pathway to play anti-tumor effects.
出处
《中国中医药信息杂志》
CAS
CSCD
2016年第10期78-81,共4页
Chinese Journal of Information on Traditional Chinese Medicine
基金
甘肃省自然科学基金(1506RJZA251)
兰州市科技计划项目(2012-1-32)
