华蟾素对人胃癌MKN-28细胞增殖、凋亡的影响研究

Study on the Effect of Cinobufotalin on the Proliferation and Apoptosis of Human Gastric Cancer MKN-28 Cells

目的:探究华蟾素对人胃癌细胞MKN-28细胞株的增殖、凋亡、细胞周期的影响,揭示华蟾素的抗癌机制。方法:选用人胃癌细胞MKN-28细胞株作为研究对象,采用不同浓度华蟾素处理细胞后,通过Cell Counting Kit-8实验(CCK8法)计算半抑制浓度(IC50),再以IC50值作为最佳药物使用浓度处理MKN-28细胞,同时以DMSO作为对照,比较两组细胞在增殖、凋亡、细胞周期等方面的差异。采用Western Blot检测两组细胞Bcl-2、BAX、Caspase-3等凋亡相关蛋白的表达情况。结果:CCK8实验结果显示,随华蟾素处理浓度的增加,胃癌细胞增殖能力逐渐下降。华蟾素对MKN-28细胞作用的IC50值为0.16μg/mL。在0.16μg/mL华蟾素作用下,对照组胃癌细胞凋亡比例为(7.87±0.50)%,而实验组凋亡比例为(16.62±1.21)%,差异具有统计学意义(P=0.002)。实验组G0/G1期细胞所占比例显著增加,而G2/M期细胞所占比例显著降低,差异具有统计学意义(P<0.05)。经过华蟾素处理后,MKN-28细胞中,Bcl-2蛋白表达量显著上升(P=0.002),而Bax、Caspase-3表达量则显著下降(P<0.05)。结论:华蟾素能使人胃癌细胞MKN-28的增殖能力下降,诱导凋亡,阻滞胃癌进展相关的细胞周期,具有作为潜在抗癌药物的开发价值。

Objective:To investigate the effects of cinobufotalin on the proliferation,apoptosis,and cell cycle of human gastric cancer cell line MKN-28,and to reveal the anticancer mechanism of cinobufalin.Methods:The human gastric cancer cell line MKN-28 was selected as the research object.After treating the cells with different concentrations of cinobufotalin,the half maximum inhibition concentration(IC50)was calculated through the Cell Counting Kit-8 experiment(CCK8 method).The IC50 value was used as the optimal drug concentration to treat MKN-28 cells,while DMSO was used as the control.The differences in proliferation,apoptosis and cell cycle between the two groups of cells were compared.Western blot was used to detect the expression of apoptosis related proteins such as Bcl-2,Bax,and caspase-3 in the two groups.Results:The results of the CCK8 experiment showed that the proliferation ability of gastric cancer cells gradually decreased with the increase of the concentration of cinobufotalin.The IC50 value of cinobufotalin on MKN-28 cells is 0.16μg/mL.Under the action of cinobufotalin with concentration of 0.16μg/mL,the proportion of gastric cancer cell with apoptosis in the control group was(7.87±0.50)%,while the proportion of apoptosis in the experimental group was(16.62±1.21)%,with a statistically significant difference(P=0.002).The proportion of G0/G1 phase cells in the experimental group significantly increased,while the proportion of G2/M phase cells significantly decreased,with a statistically significant difference(P<0.05).After treatment with cinobufotalin,the expression of Bcl-2 protein in MKN-28 cells was increased significantly(P=0.002),while the expression of Bax and Caspase-3 was decreased significantly(P<0.05).Conclusion:Cinobufotalin can reduce the proliferation ability of human gastric cancer cell line MKN-28,induce apoptosis,and block the cell cycle related to gastric cancer progression,which has potential development value as a potential anticancer drug.