去甲斑蝥素对人胰腺癌PANC-1细胞凋亡作用研究.PDF

江医学2015 年第37 卷第13 期 去甲斑蝥素对人胰腺癌PANC-1 细胞凋亡作用的研究 张日沅 林 璋 陈翀 王盈盈 徐贤绸 【摘要 】 目的 探讨去甲斑蝥素对人胰腺癌PANC-1 细胞凋亡的作用及其机制。方法 将PANC-1 细胞株随机分为处理组 和对照组，处理组加入不同浓度的去甲斑蝥素培养24h 后，CCK-8 法检测细胞增殖情况；流式细胞术分析细胞的凋亡情况；免疫印迹法 检测细胞内质网应激和凋亡相关蛋白的表达；荧光定量PCR 术检测细胞内质网应激和凋亡相关蛋白mRNA 表达。 结果 不同浓 度去甲斑蝥素处理PANC-1 细胞24h 后，与对照组相比，能降低细胞的存活率并诱导其凋亡。与对照组相比，处理组中内质网应激和凋 亡相关蛋白的表达水平均上调（均P 0.05 ），同时其mRNA 的表达水平亦均上调（均P 0.05 ）。结论 去甲斑蝥素能明显抑制人胰腺 癌PANC-1 细胞的生长并诱导其凋亡，并且具有浓度依赖性，这一作用可能是通过内质网应激介导的凋亡途径实现的。 【关键词】 胰腺癌 去甲斑蝥素 凋亡 内质网应激 Effect of norcantharidin on human pancreatic cancer PANC-1 cells , China 【Abstract 】 Objective To investigate the effect of norcantharidin on human pancreatic cancer PANC-1 cells. Methods PANC-1 cells were cultured with 0.1% DMSO and norcantharidin (0, 10, 50, 75, 100, 150, 200 mol/L) for 24h. The growth inhibitory effect was observed by using CCK-8, cell apoptosis was determined by flow cytometry (FCM). The expressions of endoplasmic reticulum stress (ERS) markers were detected by Western blot, expressions of GRP78 and GADD153 mRNA were detected by re- al-time PCR after exposure to norcantharidin. Results CCK-8 assay showed that norcantharidin significant inhibited the growth of PANC-1 cells. Apoptosis rates of PANC-1 cells treated with norcantharidin were significantly higher than those of control group. Western blot analysis showed that norcantharidin up-regulated ERS markers (P 0.05). Real-time PCR showed that the expression of GRP78 and GADD153 mRNA in PANC-1 cells treated with norcantharidin was higher than that of control group (P 0.05). Conclusion Norcantharidin can inhibit the growth of human pancreatic cancer PANC-1 cells and induce cell apoptosis. ERS-mediated apoptosis pathway may be involved in norcantharidin-induced apoptosis in pancreatic cancer PANC-1 cells. 【Key words 】 Pancreatic canc