食品科学

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山楂原花青素调节环氧合酶-2表达诱导Eca109食管癌细胞凋亡

宓 伟,练 武,尹淑英,衣卫杰,石塔拉,韩文婷   

  1. 滨州医学院公共卫生与管理学院,山东 烟台 264003
  • 出版日期:2016-01-15 发布日期:2016-01-15

Hawthorn Procyanidins Regulate the Expression of COX-2 and Induce the Apoptosis of Eca109 Esophageal Cancer Cells

MI Wei, LIAN Wu, YIN Shuying, YI Weijie, SHI Tala, HAN Wenting   

  1. School of Public Health and Management, Binzhou Medical University, Yantai 264003, China
  • Online:2016-01-15 Published:2016-01-15

摘要:

探讨山楂原花青素(hawthorn procyanidins,HPC)通过磷脂酰肌醇3-激酶/丝苏氨酸蛋白激酶(phosphatidylinositol 3-kinase/serine threonine protein kinase,PI3K/Akt)信号通路调节环氧合酶-2(cyclooxygenase-2,COX-2)表达,诱导Eca109食管癌细胞凋亡的分子机制。体外常规培养食管癌细胞株,配制25、50、100、200、300、400 μg/mL不同质量浓度的HPC,CCK-8(Cell Counting Kit-8)法检测并计算不同质量浓度HPC在72 h内对Eca109食管癌细胞的抑制率,选取IC50=250 μg/mL作为HPC处理组剂量,并设对照组。Western blotting法检测Eca109细胞PI3K和Akt磷酸化程度、Caspase-3和COX-2蛋白表达水平,反转录聚合酶链式反应(reverse transcript polymerase chain reaction,RT-PCR)检测Eca109细胞COX-2 mRNA表达水平,Annexin V-FITC/PI法检测0、12、24、36、48、72 h时HPC对Eca109食管癌细胞凋亡率的影响。结果表明:HPC处理组Eca109食管癌细胞PI3K、Akt磷酸化程度随着处理时间延长逐渐降低(P<0.01),在48 h时活性最低,HPC处理组Eca109食管癌细胞COX-2 mRNA的表达量与对照组比较明显降低(P<0.01)。在HPC的作用下,Eca109食管癌细胞Caspase-3蛋白表达量在48、72 h时最强(P<0.01),COX-2蛋白表达量在72 h时最低(P<0.01)。Eca109食管癌细胞凋亡率随着时间的延长越来越高,在72 h时达到63.54%(P<0.01)。HPC可通过PI3K/Akt途径下调COX-2表达,从而诱导Eca109食管癌细胞凋亡。

关键词: 山楂原花青素, 环氧合酶-2, 食管癌, 细胞凋亡

Abstract:

The present work was undertaken to explore the molecular mechanisms for the apoptosis of esophageal
cancer cells induced by HPC through regulating the expression of cyclooxygenase-2 (COX-2) via the PI3K/Akt signal
pathway. Esophageal cancer cells were nurtured routinely at HPC concentrations of 25, 50, 100, 200, 300 and 400 μg/mL.
The cell counting kit-8 (CCK-8) assay was used to detect and calculate the inhibitory rate of esophageal cancer cells after
72 hours of culture. The IC50 of 250 μg/mL was chosen as the experimental group. Western blotting was adopted to detect
the degree of PI3K/Akt phosphorylation and the protein expression levels of Caspase-3 and COX-2. RT-PCR was used to
evaluate the expression level of COX-2 mRNA. Annexin V-FITC/PI was used to test inhibitory rate of esophageal cancer
cells after 0, 12, 24, 36, 48 and 72 h. The degree of PI3K and Akt phosphorylation declined gradually as the culture time
elapsed and reached the lowest point (P < 0.01) after 48 hours in the experimental group, while the opposite trend was
observed for the control group (P < 0.01) . The expression of COX-2 mRNA in the experimental group was significantly
lower (P < 0.01) compared to that in the control group. Under the action of HPC, the expression of Caspase-3 protein
was the strongest (P < 0.01) after 48 and 72 hours while the expression of COX-2 protein was the lowest (P < 0.01) after
72 hours. The apoptosis rate of esophageal cancer cells increased as the culture period was prolonged, which reached
63.54% (P < 0.01) after 72 hours of culture. HPC can exert a pro-apoptotic effect on esophageal cancer cells by reducing the
expression level of COX-2 through the PI3K/Akt signal pathway.

Key words: hawthorn procyanidins, cyclooxygenase-2, esophageal cancer, cell apoptosis

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