摘要目的 观察鞣花酸(EA)对胰腺癌细胞株PANC-1增殖、凋亡的影响,探讨鞣花酸诱导PANC-1凋亡的机制.方法 用不同浓度的鞣花酸(0、2、4、6、8μg/ml)干预PANC-1细胞不同时间(24、48、72 h)后,采用倒置显微镜观察细胞形态变化,噻唑蓝(MTT)比色法检测细胞增殖.取0、2、4、6 μg/ml的鞣花酸干预PANC-1细胞48 h后,细胞染色观察细胞凋亡的形态改变,流式细胞仪检测细胞凋亡率和周期,Western blot法检测环氧合酶-2(COX-2)和核因子-κB (NF-κB)蛋白的表达.结果 鞣花酸对PANC-1细胞有明显的生长抑制作用,呈现明显的浓度依赖性和时间依赖性,不同浓度组、不同时间组间增殖抑制率差异有统计学意义(P＜0.05),且加药组贴壁细胞出现皱缩、变圆、脱落.不同浓度的鞣花酸干预PANC-1细胞48 h后,细胞形态发生典型凋亡特征性改变,细胞凋亡率分别为(11.18 ±0.99)％、(20.70 ±2.32)％、(39.68 ±0.69)％,与对照组凋亡率[(0.62±0.23)％]比较,不同浓度差异有统计学意义(P＜0.05);G0/G1期细胞百分比分别为(48.98±2.36)％、(60.80±1.40)％、(72.82 ±0.59)％,与对照组[(42.78±O.74)％]比较,不同浓度差异有统计学意义(P＜0.05);Western blot检测提示COX-2和NF-κB的蛋白表达明显降低,且呈浓度依赖性.结论 鞣花酸可抑制胰腺癌细胞PANC-1的增殖、诱导其凋亡,其机制可能与阻滞细胞周期,下调COX-2和NF-κB的蛋白表达有关.

abstractsObjective To investigate the effect of ellagic acid on the proliferation and apoptosis of pancreatic cell line PANC-1 in vitro and explore possible mechanism of ellagic acid' s effect on PANC-1 cells.Methods PANC-1 cells were treated with ellagic acid at concentrations of 0,2,4,6,8 μg/ml for 24,48 and 72 h respectively.Inverted Microscope showed cells' morphological changes.The cell proliferation was determined by methyl thiazolyl tetrazolium (MTT) assay.When the cells were treated with ellagic acid at concentrations of 0,2,4,6 μg/ml for 48 h,the morphological changes of apoptotic cells were observed by cell staining,apoptotic rate and cell cycle were detected by flow cytometry,the protein expression of cyclo-oxgenase 2 (COX-2) and nuclear factor-κB (NF-κB) were measured by Western blotting.Results Ellagic acid could significantly inhibited the growth of PANC-1 cells in a concentration-dependent and time-dependent manner.The difference among these groups were statistically significant(P ＜0.05).After treatment with ellagic acid,apoptosis of PANC-1 cells was seen.Apoptosis rates in the different concentration groups were respectively (11.18 ± 0.99) ％,(20.70 ± 2.32) ％,(39.68 ± 0.69) ％,the apoptosis rate of control group [(0.62 ± 0.23) ％] compared with statistically significant difference among the different concentration groups (P ＜ 0.05).The percentages of G0/G1 phase cells were respectively(48.98±2.36)％,(60.80 ± 1.40)％,(72.82 ±0.59)％,the percentage of control group [(42.78 ±0.74)％] compared with statistically significant difference among the different concentration groups(P ＜0.05).The protein expression of COX-2 and NF-κB measured by Western blotting were significantly down-regulated dose-dependently.Conclusion Ellagic acid can inhibit the proliferation of pancreatic cancer cell line PANC-1 and induce apoptosis.The mechanism may depend on arresting cell cycle and decreasing protein expression of COX-2 and NF-κB.