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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (3): 262-265.doi: 10.12047/j.cjap.6108.2021.004

• 研究论文 • 上一篇    下一篇

川楝素诱导人胃癌MGC-803细胞凋亡及其机制

徐兴军1,2, 李珊珊1, 刘畅1, 张伟伟1,2, 张珍珠1,2, 邵淑丽1,2Δ   

  1. 1. 齐齐哈尔大学生命科学与农林学院, 黑龙江 齐齐哈尔 161006;
    2. 抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江 齐齐哈尔 161006
  • 出版日期:2021-05-28 发布日期:2021-08-09
  • 通讯作者: Tel: 13836267458; E-mail: shshl32@163.com
  • 基金资助:
    *齐齐哈尔大学黑龙江省教育厅基本业务专项重点项目(135109104);黑龙江省高教强省优势特色学科—玉米“粮头食尾”重点专项(LTSW201737);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(YSTSXK201809)

Toosendanin induces apoptosis of human gastric cancer MGC-803 cells and its mechanism

XU Xing-jun 1,2, LI Shan-shan1, LIU Chang 1, ZHANG Wei-wei 1,2, ZHANG Zhen-zhu 1,2, SHAO Shu-li 1,2Δ   

  1. 1. College of Life Science and Agriculture and Forestry, Qiqihar University, Qiqihaer 161006;
    2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar University, Qiqihaer 161006, China
  • Online:2021-05-28 Published:2021-08-09

摘要: 目的: 本研究旨在探讨川楝素诱导人胃癌MGC-803细胞凋亡及其机制。方法: 将人胃癌MGC-803细胞分为5组,每组3个复孔,采用氟尿嘧啶(5-FU)和0 nmol/L川楝素(TSN)分别作为阳性对照和阴性对照。其余3组分别加入终浓度为30 nmol/L、50 nmol/L、70 nmol/L的川楝素。川楝素处理细胞48 h后,利用激光共聚焦显微镜观察细胞形态结构变化;流式细胞术检测线粒体膜电位变化;酶标法检测Caspase-3和Caspase-9活性;利用qRT-PCR和Western blot检测凋亡相关基因Bcl-2BaxCyt cAPAF-1 mRNA和蛋白水平。结果: 与0 nmol/L TSN组相比,30 nmol/L、50 nmol/L、70 nmol/L的川楝素作用于人胃癌MGC-803细胞48 h,可见细胞体积缩小,细胞核裂解,部分染色质凝集等形态学变化;Caspase-3和Caspase-9活性升高(P<0.05);而线粒体膜电位明显下降(P< 0.05);BaxCyt cAPAF-1 基因mRNA及蛋白表达量显著升高(P<0.05),Bcl-2 基因mRNA及蛋白表达量显著降低(P<0.05)。结论: 川楝素通过上调BaxCyt cAPAF-1的表达,下调Bcl-2基因表达,增强Caspase-3、Caspase-9活性诱导人胃癌MGC-803细胞凋亡。

关键词: 川楝素, MGC-803细胞, 细胞凋亡

Abstract: Objective: To explore the apoptosis of human gastric cancer MGC-803 cells induced by toosendanin(TSN) and its mechanism. Methods: The human gastric cancer MGC-803 cells were divided into 5 groups, each group was set with 3 replicate. Fluorouracil (5-FU) and 0 nmol/L toosendanin (TSN) were used as positive control and negative control, respectively. The other three groups were treated with toosendanin at the final concentrations of 30, 50, and 70 nmol/L, respectively. After 48 h of treatment with toosendanin, the morphology of the cells were observed under laser confocal microscopy. Flow cytometry was used to detect the mitochondrial membrane potential, and enzyme-labeled assays were used to detect the activities of Caspase-3 and Caspase-9. The mRNA and protein levels of Bcl-2, Bax, Cyt c and APAF-1 were measured by qRT-PCR and Western blot. Results: Compared with the 0 nmol/L TSN group, after the human gastric cancer MGC-803 cells were treated with toosendanin at the concentrations of 30, 50, and 70 nmol/L for 48 h, the cell volume shrinkage, nucleus cleavage and chromatin morphological changes were observed under the microscope. The activities of Caspase-3 and Caspase-9 were increased significantly (P<0.05), while the mitochondrial membrane potential was decreased significantly (P<0.05). In addition, the mRNA and protein expression levels of Bax, Cyt c and APAF-1 were increased significantly (P<0.05), while the mRNA and protein expression levels of Bcl-2 were decreased significantly (P<0.05). Conclusion: Toosendanin up-regulates the expressions of Bax, Cyt c and APAF-1, down-regulates the expression of Bcl-2 gene, enhances the activities of caspase-3 and caspase-9, and induces the apoptosis of human gastric cancer MGC-803 cells.

Key words: toosendanin, MGC-803cells, cell apoptosis

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