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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (5): 514-519.doi: 10.12047/j.cjap.6122.2021.069

• 研究论文 • 上一篇    下一篇

大蒜阿霍烯抗胃癌的作用及分子机制*

朱明了+, 郑珍+, 楼恩哲+, 赵凯婷, 何施燕, 陈佳玉   

  1. 绍兴文理学院医学院, 浙江 绍兴 312000
  • 收稿日期:2020-07-10 修回日期:2021-01-22 出版日期:2021-09-28 发布日期:2021-11-24
  • 通讯作者: Tel: 0575-88345826; E-mail: chenyujia10@163.com
  • 作者简介:+: 为共同第一作者
  • 基金资助:
    *国家大学生创新创业训练计划项目(201910349030,202010349038);浙江省大学生科技创新活动计划项目(2020R444013)

Anti-gastric cancer effects of Z Ajoene and its molecular mechanisms

ZHU Ming-liao+, ZHENG Zhen+, LOU En-zhe+, ZHAO Kai-ting, HE Shi-yan, CHEN Jia-yu   

  1. Medical school, Shaoxing University, Shaoxing 312000, China
  • Received:2020-07-10 Revised:2021-01-22 Online:2021-09-28 Published:2021-11-24

摘要: 目的: 探讨大蒜阿霍烯对胃癌细胞MGC-803的作用及相关分子机制。方法: 终浓度分别为0、1、5、25和125 μmol/L大蒜阿霍烯作用于细胞MGC-803 24 h、48 h和72 h,各设3复孔,MTS法检测细胞增殖活性,JC-1和Hoechst染色法观察线粒体膜电位和核型改变,LDH释放法检测细胞毒性,流式法分析细胞的凋亡和周期改变,RT-qPCR和Western blot检测P53、Caspase-3、RAS、ERK、BCL-2、AKT、mTOR、PI3K基因的表达,同时,取4周龄雄性BALB/C鼠随机分成5组,20只/组,腹股沟皮下接种胃癌细胞MGC-803,2 d后各组分别经皮下注射浓度为0 μmol/L、1 μmol/L、5 μmol/L、25 μmol/L、125 μmol/L的大蒜阿霍烯,0.1 ml/次,隔天注射一次,并于首次注射肿瘤细胞的第20日每组杀10只,取瘤组织,称重。记录剩余小鼠的存活期,观察大蒜阿霍烯对荷瘤小鼠胃癌生长及生存期的影响。结果: 大蒜阿霍烯可明显抑制MGC-803细胞增殖活性,诱导细胞凋亡(P<0.01),显著上调P53、Caspase-3、BAX基因的转录和表达水平,抑制基因RAS、ERK1、BCL-2、AKT、mTOR和PI3K的表达(P<0.01),显著抑制小鼠胃癌移植瘤生长,并延长荷瘤小鼠存活期(P<0.01)。结论: 大蒜阿霍烯对胃癌有治疗作用,可通过调节PI3K-AKT-mTOR、RAS-RAF-MEK-ERK信号通路而抑制胃癌细胞增殖,诱导细胞凋亡。

关键词: 大蒜阿霍烯, 胃癌细胞, 凋亡, 信号通路, 小鼠

Abstract: Objective: To investigate the effects of Z Ajoene on gastric cancer cell MGC-803 and its molecular mechanisms. Methods: The gastric cancer cells MGC-803 were treated with 0, 1, 5, 25 and 125 μmol/L Z Ajoene for 24 h, 48 h and 72 h, each with 3 replicate wells. The proliferation activity of MGC-803 cells was analyzed by MTS method, mitochondrial membrane potential was analyzed after JC-1 staining, nuclear type was observed after Hoechst 33342 staining, cytotoxicity was detected by LDH release method, and the apoptosis level and cell cycle were analyzed with flow cytometry. RT-qPCR and Western blot methods were used to evaluate the expression levels of P53, Caspase-3, RAS, ERK, BCL-2, AKT, mTOR and PI3K genes. At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, 20 per group, and were subcutaneously inoculated with gastric cancer cell MGC-803 in the groin. Two days later, each group was injected with Z Ajoene at the doses of 0, 1, 5, 25 and 125 μmol/L, 0.1 ml/time, and was injected every other day. On the 20th day of the first injection of tumor cells, 10 mice in each group were killed, the tumor tissues were taken out and weighed. The survival period of the remaining mice was recorded and the effects of Z Ajoene on the growth and survival period of gastric cancer in tumor-bearing mice were observed. Results: After Z Ajoene treatment, the proliferation activity of MGC-803 cells was significantly inhibited and the apoptosis rate was significantly increased(P<0.01). The transcription and expression levels of p53, Caspase-3 and BAX genes were significantly increased, while the transcription and expression levels of RAS, ERK1, BCL-2, AKT, mTOR and PI3K genes were decreased markedly(P<0.01). The tumor inhibition experiments showed that the growth of the tumor could be inhibited and the survival time of the tumor-bearing animals could be greatly prolonged after Z Ajoene treatment(P<0.01). Conclusion: Z Ajoene has therapeutic effects on gastric cancer, can inhibit the proliferation of gastric cancer cells and induce them apoptosis by regulating the expression of PI3K-AKT-mTOR and RAS-RAF-MEK-ERK signal pathways.

Key words: Z Ajoene, gastric cancer cells, apoptosis, signal pathway, mouse

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