β―榄香烯对人胰腺癌Panc―1细胞凋亡影响.docVIP

β―榄香烯对人胰腺癌Panc―1细胞凋亡影响 　　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、160 μg/mL，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、FasL、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（P0.05，P0.01），细胞凋亡率明显增加（P0.01，P0.001），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（P0.05，P0.01），Fas、FasL、Cyt c及AIF蛋白表达明显增强（P0.05，P0.01，P0.001）。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。 　　关键词：β-榄香烯；胰腺癌；Panc-1细胞；细胞凋亡 　　DOI：10.3969/j.issn.1005-5304.2016.10.018 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）10-0078-04 　　Abstract： 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/mL） were incubated to the Panc-1 cells in vitro cultured for 24 h， 48 h and 72h， 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， FasL and Cyt c and AIF. Results The activity of Panc-1 cells was obviously inhibited time/concentration dependent inhibition （P0.05， P0.01）， and the apoptosis rate increased after incubated with β-Elemene （P0.01， 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 （P0.05， P0.01）； protein expressions of Fas， FasL， Cyt c and AIF were significantly enhanced （P0.05， P0.01， P0.001）. Conclusion β-Elemene can