Wang Qi, 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 161006, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2021 May;37(3):254-256. doi: 10.12047/j.cjap.6054.2021.001.
In this study, human gastric cancer MGC-803 cells were treated with betulinic acid at different concentrations to investigate its effect on cell autophagy. The human gastric cancer MGC-803 cells were divided into 4 groups, each group was set with 3 replicate. The control group was not treated with betulinic acid, the other three groups were added with final concentration of 10,20,30 mg/L betulinic acid, respectively. Cells were treated with betulinic acid for 48 h,qRT-PCR was applied to detect the effect of betulinic acid on mRNA expressions of autophagy-related genes in human gastric cancer MGC-803 cells. Western blot was performed to determine the protein expressions of cell autophagy-related genes after drug treatment. Immunofluorescence was used to detect the localization and expression of LC3 protein in MGC-803 cells after drug treatment. Compared with the control group,in the concentration range of 1030 mg/L, the mRNA expression of LC3 and Beclin-1 in human gastric cancer MGC-803 cells treated with betulinic acid were increased significantly, the expressions of Beclin-1 and LC3-Ⅱ protein were also increased significantly, while the expression of LC3-Ⅰ protein was decreased significantly. Among them, betulinic acid at the concentration of 30 mg/L showed the best effects. In addition, betulinic acid induced the LC3 protein in MGC-803 cells to form spot aggregates in the cytoplasm. At the concentrations of 1030 mg/L, betulinic acid can induce autophagy in human gastric cancer MGC-803 cells.
在本研究中，将人胃癌MGC - 803细胞用不同浓度的桦木酸处理，以研究其对细胞自噬的影响。将人胃癌MGC - 803细胞分为4组，每组设3个重复。对照组不进行桦木酸处理，其他三组分别加入终浓度为10、20、30 mg/L的桦木酸。用桦木酸处理细胞48 h后，采用qRT - PCR检测桦木酸对人胃癌MGC - 803细胞自噬相关基因mRNA表达的影响。进行蛋白质免疫印迹法以测定药物处理后细胞自噬相关基因的蛋白质表达。采用免疫荧光法检测药物处理后MGC - 803细胞中LC3蛋白的定位和表达。与对照组相比，在1030 mg/L浓度范围内，用桦木酸处理的人胃癌MGC - 803细胞中LC3和Beclin - 1的mRNA表达显著增加，Beclin - 1和LC3 -Ⅱ蛋白的表达也显著增加，而LC3 -Ⅰ蛋白的表达显著降低。其中，30 mg/L浓度的桦木酸效果最佳。此外，桦木酸诱导MGC - 803细胞中的LC3蛋白在细胞质中形成斑点状聚集物。在1030 mg/L浓度下，桦木酸可诱导人胃癌MGC - 803细胞发生自噬。
