低浓度乙醇诱导人胰腺癌细胞凋亡的实验研究

目的:探讨低浓度乙醇对胰腺癌细胞凋亡的诱导作用。方法:用含不同浓度乙醇（0%、2%、4%和6%）培养液培养胰腺癌细胞。CCK8法检测不同浓度乙醇对胰腺癌细胞增殖的影响，流式细胞仪检测细胞的凋亡。实时荧光定量PCR法检测凋亡相关基因caspase-8 mRNA、caspase-9 mRNA的表达水平变化，Western blot检测caspase-3蛋白表达水平的变化。结果:低浓度乙醇可抑制胰腺癌细胞的增殖，随着浓度的增加，细胞存活率明显下降,与对照组（0%组）相比均有统计学差异(P<0.05)。流式细胞仪检测显示2%、4%和6%乙醇组均可诱导细胞凋亡，与对照组（0%组）相比均有统计学差异(P<0.05)。caspase-8 mRNA在2%、4%和6%乙醇组的表达与对照组相比未见明显增加（P>0.05），而caspase-9 mRNA在2%、4%和6%乙醇组中的表达均增加，且与对照组相比均有统计学差异（P<0.05）。活性caspase-3蛋白在2%、4%和6%乙醇组中均有表达，而对照组未见活性caspase-3蛋白的表达。结论:低浓度乙醇可诱导胰腺癌细胞凋亡。

Low concentrations of ethanol induce apoptosis in human pancreatic cancer cell lines

Objective:To determine the effect of low concentrations of ethanol on apoptosis of cells in a human pancreatic cancer cell lines.Methods:Pancreatic cancer cells were incubated in DMEM with ethanol concentrations （2%，4% and 6%) or without ethanol concentrations （0%，control group) for 6h.Cell proliferation rate was measured by means of CCK8.Apoptosis was assessed by means of flow cytometry.Caspase-8 and caspase-9 mRNA were analyzed by real-time quantitative PCR and caspase-3 protein was analyzed by Western blot. Results:Cell proliferation was inhibited in groups of 2%，4% and 6% ethanol concentrations compared with control group （P<0.05).As the concentration increased，the rate of cell proliferation decreased significantly.Flow cytometry showed that 2%，4% and 6% ethanol group can induce apoptosis of pancreatic cancer cells.There was statistically significance compared with to the control group （0%) （P<0.05).Expression of caspase-8 mRNA didn' t increased significantly in groups of 2%，4% and 6% ethanol concentrations compared with control group （P>0.05).caspase-9 mRNA expression increased markedly in groups of 2%，4% and 6% ethanol concentrations compared with control group （P<0.05).Caspase-3 protein expression increased significantly in groups of 2%，4% and 6% ethanol concentrations，whereas wasn' t observed in control group.Conclusion:Low ethanol concentrations can induce apoptosis of pancreatic cancer cells.