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川楝素通过死亡受体途径诱导人胃癌MGC-803细胞凋亡

Toosendan Induces Apoptosis of Human Gastric Cancer MGC-803 Cells Through the Death Receptor Pathway

  • 摘要: 为探讨川楝素(toosendanin, TSN)诱导人胃癌MGC-803细胞凋亡及其机制,为川楝素治疗胃癌提供参考依据。本研究采用氟尿嘧啶和0 nmol/L TSN分别作为阳性对照和阴性对照。以终浓度为30、50、70 nmol/L的TSN作用于人胃癌MGC-803细胞48 h,采用激光共聚焦显微镜观察细胞形态结构,流式细胞术检测细胞凋亡率,酶标法检测Caspase-3和Caspase-8活性,RT-qPCR和Western blot检测Fas、Caspase-3和Caspase-8基因表达水平。与0 nmol/L TSN组相比,30、50、70 nmol/L的TSN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;早期凋亡细胞和晚期凋亡细胞所占百分比均明显增加;Caspase-3和Caspase-8活性显著升高;Fas、Caspase-3和Caspase-8基因m RNA及蛋白表达水平明显升高。综上所述,川楝素可通过死亡受体途径诱导人胃癌MGC-803细胞凋亡。

     

    Abstract: The purpose of this study was to investigate the apoptosis of human gastric cancer MGC-803 cells induced by toosendanin and its mechanism, so as to provide reference for toosendanin in the treatment of gastric cancer. Fluorouracil and 0 nmol/L toosendanin(TSN) were used as positive control and negative control, respectively.Human gastric cancer MGC-803 cells were treated with TSN at a final concentration of 30 nmol/L, 50 nmol/L,and 70 nmol/L for 48 h, and the morphological structure of the cells was observed by laser confocal microscopy.Flow cytometry was used to detect the apoptosis rate, enzyme-labeled method was used to detect the activity of Caspase-3 and Caspase-8, RT-qPCR and Western blot were used to detect the gene expression level of Fas, Caspase-3 and Caspase-8. Compared with the 0 nmol/L TSN group, 30 nmol/L, 50 nmol/L, and 70 nmol/L TSN acted on human gastric cancer MGC-803 cells for 48 h, showing reduced cell size, nuclear lysis, and partial chromatin Morphological changes such as agglutination. The percentage of early apoptotic cells and late apoptotic cells increased significantly; the activity of Caspase-3 and Caspase-8 increased significantly; the mRNA and protein expression levels of Fas, Caspase-3 and Caspase-8 genes increased significantly. In summary, toosendanin can induce apoptosis of human gastric cancer MGC-803 cells through the death receptor pathway.

     

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