Fan Hua, Shao Shu Li, He Meng Qi, Huang Xin, Zhang Wei Wei, Zhang Zhen Zhu
College of Life Science and Agriculture and Forestry, Qiqihar University, Qiqihaer 161000.
Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar University, Qiqihaer 161000, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2021 Jul;37(4):419-422. doi: 10.12047/j.cjap.6078.2021.039.
The effects of betulinic acid (BA) on apoptosis of human gastric cancer MGC-803 cells was investigated by using human gastric cancer MGC-803 cells as experimental materials, and the basis for its clinical application was provided. The human gastric cancer MGC-803 cells were divided into 4 groups,each group was set with 3 replicates.The control group was MGC-803 cells without being added betulinic acid; the other 3 groups of experimental groups were treated with betulinic acid at final concentrations of 10, 20 and 30 μg /ml respectively. Cells were treated with betulinic acid of different concentrations for 48 h. Laser confocal microscope was used to observe morphological changes of MGC-803. The activities of Caspase-3 and Caspase-9 were detected by an assay kit. Flow cytometry was applied to determine mitochondrial membrane potential. The mRNA and protein levels of and were also detected by qRT-PCR and Western blot, respectively. Compared with the control group, the activities of Caspase-3 and caspase-9 were increased(<0.01), while the mitochondrial membrane potential was decreased significantly(<0.01). The mRNA and protein expressions of Caspase-3, caspase-9 and Cyt c were up-regulated significantly(<0.01). In the final concentration range of 10 ~ 30 μg/ml, betulinic acid can induce apoptosis of human gastric cancer MGC-803 cells by regulating the expression of Caspase-3, Caspase-9 and Cyt c.
以人胃癌MGC-803细胞为实验材料,研究了桦木酸(BA)对人胃癌MGC-803细胞凋亡的影响,为其临床应用提供依据。将人胃癌MGC-803细胞分为4组,每组设3个重复。对照组为未添加桦木酸的MGC-803细胞;其他3组实验组分别用终浓度为10、20和30μg/ml的桦木酸处理。用不同浓度的桦木酸处理细胞48小时。采用激光共聚焦显微镜观察MGC-803的形态变化。用试剂盒检测Caspase-3和Caspase-9的活性。应用流式细胞术检测线粒体膜电位。同时分别用qRT-PCR和Western blot检测Caspase-3、Caspase-9和细胞色素c(Cyt c)的mRNA和蛋白水平。与对照组相比,Caspase-3和Caspase-9的活性升高(P<0.01),而线粒体膜电位显著降低(P<0.01)。Caspase-3、Caspase-9和Cyt c的mRNA和蛋白表达均显著上调(P<0.01)。在10~30μg/ml的终浓度范围内,桦木酸可通过调节Caspase-3、Caspase-9和Cyt c的表达诱导人胃癌MGC-803细胞凋亡。