胡椒碱诱导人胰腺癌PANC-1细胞凋亡的Caspase 3/Bax/Bcl-2信号通路机制研究

Study on the Caspase 3/Bax/Bcl-2 Signal Pathway Mechanism of Induction Apoptosis Effect of Piperine in Human Pancreatic Cancer PANC-1 Cell

关键词:  胡椒碱  人胰腺癌  PANC-1细胞  凋亡  Caspase 3/Bax/Bcl-2

Key words:  piperine  human pancreatic cancer  PANC-1 cell  apoptosis  Caspase 3/Bax/Bcl-2

摘要:

目的 观察胡椒碱对人胰腺癌PANC-1细胞增殖、凋亡的影响，探讨其诱导凋亡的Caspase 3/Bax/Bcl-2信号通路机制。方法 将人胰腺癌PANC-1细胞分为阴性对照组和40，20，10 μmol·L^-1胡椒碱组，采用MTT、台盼蓝染色计数及平板克隆形成试验法检测胡椒碱对PANC-1细胞增殖、生长曲线及克隆形成的影响；采用Hoechst 33258染色法观察胡椒碱对PANC-1细胞凋亡形态学的影响；采用RT-PCR和Western blotting法检测胡椒碱对PANC-1细胞Caspase 3、cleaved-Caspase 3、Bax及Bcl-2 mRNA及蛋白表达的影响。结果 MTT、台盼蓝染色计数和平板克隆形成实验结果显示40，20 μmol·L^-1胡椒碱可明显抑制PANC-1增殖、细胞生长曲线和克隆形成；Hoechst 33258染色实验显示40，20，10 μmol·L^-1胡椒碱有诱导PANC-1细胞凋亡作用；RT-PCR和Western blotting实验结果显示40，20 μmol·L^-1胡椒碱可上调PANC-1细胞Caspase 3、Bax mRNA表达水平，上调cleaved-Caspase 3和Bax蛋白表达水平，下调Bcl-2 mRNA和蛋白表达水平。结论 胡椒碱可抑制人胰腺癌PANC-1细胞生长、增殖，并诱导其凋亡，其机制可能与调控Caspase 3/Bax/Bcl-2凋亡信号通路有关。

Abstract:

OBJECTIVE To observe the effect of piperine on proliferation and apoptosis of human pancreatic cancer PANC-1 cell, and to explore its mechanism of Caspase 3/Bax/Bcl-2 signaling pathway. METHODS The human pancreatic cancer PANC-1 cells were divided into negative control group and 40, 20 and 10 μmol·L^-1 piperine groups. MTT assay, Trypan blue staining counting method and plate colony formation assays were used to test the effects of piperine on proliferation, growth curve of cells and colony formation. Hoechst 33258 staining was used to detect the effects of piperine on apoptosis of PANC-1 cells. Reverse transcription polymerase chain reaction(RT-PCR) and Western blotting were used to detect the effects of piperine on the expression levels of Caspase 3, cleaved-Caspase 3, Bax and Bcl-2 mRNA and protein in PANC-1 cells. RESULTS MTT, Trypan blue staining and plate colony formation assays showed that 40 and 20 μmol·L^-1 piperine could inhibit the proliferation, growth curve of cells and colony formation of PANC-1 cells. Hoechst 33258 staining showed that piperine 40, 20, 10 μmol·L^-1 could induce the apoptosis of PANC-1 cells. The results of RT-PCR and Western blotting showed that piperine 40, 10 μmol·L^-1 could up-regulate the expression of Caspase 3, Bax mRNA, up-regulate the expression of cleaved-Caspase 3 and Bax protein, and down-regulate the expression of Bcl-2 mRNA and protein in PANC-1 cells. CONCLUSION Piperine can inhibit the growth, proliferation and induce apoptosis in human pancreatic cancer PANC-1 cells, and its mechanism may be related to the regulation of the Caspase 3/Bax/Bcl-2 signaling pathway.