Chen Ya-Nan, 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. 2020 Nov;36(6):628-632. doi: 10.12047/j.cjap.6036.2020.131.
Human gastric cancer SGC-7901 cells were treated with betulinic acid(BA)at the concentrations of 0, 10, 20, and 30 μg/ml, and treated with conventional chemotherapeutic drug 5-Fu as a positive control to explore its effect on cell proliferation. Trypan blue and GIEMSA staining method were used to investigate the effect of BA on cell growth inhibition and clone formation. EdU method and flow cytometry were used to explore the proliferation and cell cycle of SGC-7901 cells after treated with BA, respectively. qRT-PCR and Western blot were also applied to determine the mRNA and protein levels of cyclin D1 and cyclin B1. The cell growth inhibition rate was increased after treated with different concentrations of BA in SGC-7901 cells(<0.05). After treated for 48 h, BA decreased the clone information and cell proliferation of SGC-7901 cells markedly in dose-and time-dependent manners (<0.01). Flow cytometry analysis showed that BA obviously increased the proportion of SGC-7901 cells in G1 phase but decreased the proportion of those in S phase. qRT-PCR and Western blot analysis showed that the mRNA and protein levels of cyclin D1 and cyclin B1 were significantly downregulated by BA at different concentrations(<0.01). Compared with the 5-Fu control group, when the concentration of BA was 20 μg/ml and 30 μg/ml, the cell proliferation ability was significantly decreased, the cell cycle was inhibited, and the expression of cyclin was reduced (all <0.05). The betulinic acid regulates the proliferation of SGC-7901 cells by inhibiting the expressions of cyclin D1 and cyclin B1, which leads to cell cycle arrest and proliferative inhibition.
将人胃癌SGC - 7901细胞分别用浓度为0、10、20和30μg/ml的桦木酸(BA)处理,并用传统化疗药物5 - Fu作为阳性对照来探究其对细胞增殖的影响。采用台盼蓝和吉姆萨染色法研究BA对细胞生长抑制和克隆形成的影响。分别采用EdU法和流式细胞术探究BA处理后SGC - 7901细胞的增殖和细胞周期情况。还应用qRT - PCR和蛋白质印迹法测定细胞周期蛋白D1和细胞周期蛋白B1的mRNA和蛋白质水平。不同浓度的BA处理SGC - 7901细胞后,细胞生长抑制率升高(P<0.05)。处理48小时后,BA以剂量和时间依赖性方式显著降低SGC - 7901细胞的克隆形成和细胞增殖(P<0.01)。流式细胞术分析表明,BA明显增加了SGC - 7901细胞在G1期的比例,但降低了S期细胞的比例。qRT - PCR和蛋白质印迹分析表明,不同浓度的BA均显著下调细胞周期蛋白D1和细胞周期蛋白B1的mRNA和蛋白质水平(P<0.01)。与5 - Fu对照组相比,当BA浓度为20μg/ml和30μg/ml时,细胞增殖能力显著降低,细胞周期受到抑制,细胞周期蛋白的表达降低(均P<0.05)。桦木酸通过抑制细胞周期蛋白D1和细胞周期蛋白B1的表达来调节SGC - 7901细胞的增殖,从而导致细胞周期停滞和增殖抑制。