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中国癌症防治杂志

中国癌症防治杂志 ›› 2016, Vol. 8 ›› Issue (2): 73-77.doi: DOI：10.3969/j.issn.1674-5671.2016.02.01

• 基础研究 •    下一篇

吗啡对人食管癌细胞Ec109生长及p53、caspase-3 基因表达的影响

  

  1. 广西医科大学附属肿瘤医院麻醉科
  • 出版日期:2016-04-25 发布日期:2016-05-13
  • 通讯作者: 潘灵辉 plinghui@hotmail.com
  • 基金资助:

    广西科学研究与技术开发计划资助项目（桂科攻1355005-3-16）

Effect of Morphine on growth of human esophageal cancer cell Ec109 and expression of p53 and caspase-3

  • Online:2016-04-25 Published:2016-05-13

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摘要：

目的 研究吗啡对人食管癌Ec109细胞生长及其可能的作用机制。方法 取对数生长期食管癌Ec109细胞，随机分为实验组和对照组，实验组按照吗啡不同作用浓度分为3个亚组：M1组（0.1 μmol/L）、M2组（10 μmol/L）、M3组（1 000 μmol/L）；对照组加入等容量的RPMI-l640培养基。各组分别作用24 h、48 h和72 h后，采用CCK-8法检测细胞增殖情况；作用48 h后，用流式细胞仪检测细胞周期和细胞凋亡率，RT-PCR测定p53和 caspase-3基因的表达。结果 与对照组比较，实验组M1组、M2组、M3组细胞增殖抑制率均显著升高（P＜0.01），且随吗啡浓度增加和时间延长，各实验组细胞增殖抑制率增加，呈时间和浓度依赖性（P＜0.01）；各实验组细胞凋亡率和G1期细胞比例均明显升高（P＜0.05），且随吗啡作用浓度增加而升高明显，呈浓度依赖性（P＜0.05），各实验组细胞p53 mRNA和caspase-3 mRNA的表达量均显著升高（P＜0.05），且随吗啡作用浓度的增加而升高，呈浓度依赖性（P＜0.05）。结论 吗啡可抑制食管癌Ec109细胞生长并促进其凋亡，其作用机制可能与促进p53和caspase-3基因表达有关。

关键词: 食管肿瘤, 吗啡, Ec109细胞, 增殖, 凋亡, p53；caspase-3

Abstract:

Objective To explore the effects of morphine on growth of human esophageal cancer cell Ec109 and elucidate its possible molecular mechanism. Methods Ec109 cells were randomly divided into four groups: three groups of cells were treated with different concentrations of morphine （0.1，10 and 1 000 umol/L），and a control group was treated with normovolemic RPMI-l640 nutrient solution. Cells were incubated for 24，48 and 72 h，and rates of proliferation inhibition were determined by CCK-8 assay. Flow cytometry was performed after 48-h incubation to assess apoptosis and cell cycle distribution in the four groups. We also quantified the levels of p53 and caspase-3 mRNA using real-time PCR. Results Rates of proliferation inhibition were significantly higher in the three morphine-treated groups than in the control group（P<0.01）；they were higher with higher morphine concentration and longer incubation（P<0.05）. Proportions of apoptotic cells and cells in G1 phase were also higher in the morphine-treated cells at 48 h（P<0.05），and these proportions were higher with increasing morphine concentration（P<0.05）. Levels of p53 and caspase-3 mRNAs were also up-regulated by morphine in a concentration-dependent manner（P<0.05）. Conclusion These results suggest that morphine can inhibit proliferation of Ec109 cells by inducing apoptosis. Morphine may exert these effects，in part，by up-regulating  p53 and caspase-3 expression.

Key words: Esophageal neoplasms, Morphine, Ec109 cell, Proliferation, Apoptosis, p53, Caspase-3