摘要
为明确华蟾素诱导人胃癌MGC-803细胞凋亡及其作用机制,本研究采用不同浓度华蟾素作用人胃癌MGC-803细胞48 h。激光共聚焦显微镜下观察细胞形态变化,流式细胞术检测细胞凋亡率和线粒体膜电位(mitochondrial membrane potential,MMP)变化。RT-qPCR和Western blot检测Bax、Bcl-2、Cyt-c、Caspase9、Caspase 3基因mRNA和蛋白的表达水平。结果显示,华蟾素作用人胃癌MGC-803细胞48 h后,随着药物浓度的增加,细胞呈现明显的凋亡形态,与对照组相比,线粒体膜电位显著降低,早期凋亡细胞和晚期凋亡细胞比例均显著增加。Bax、Cyt-c、Caspase 9、Caspase 3基因mRNA和蛋白水平均显著升高(P<0.05),Bcl-2基因m RNA和蛋白水平均显著降低(P<0.05)。本研究结果表明,在一定浓度范围内,华蟾素可通过上调Bax、Cyt-c、Caspase 9、Caspase 3基因表达,下调Bcl-2基因诱导人胃癌MGC-803细胞凋亡。
To investigate the effect of cinobufacin on cell apoptosis of human gastric cancer MGC-803 cells,and explore its molecular mechanisms,the human gastric cancer MGC-803 cells was treated with different concentrations of cinobufacin for 48 h.The changes of cell morphology were observed under the laser scanning confocal microscope,the rate of apoptosis and mitochondrial membrane potential were detected by the flow cytometry,the expressions of Bax,Bcl-2,Cyt-c,Caspase 9 and Caspase 3 gene m RNA and proteins were tested by RT-qPCR and Western Blot.The results showed that after 48 h of human gastric cancer MGC-803 cells treated with cinobufacin,the cells showed obvious apoptosis morphology with the increase of drug concentration,compared with the control group,the mitochondrial membrane potential is significantly reduced,and the early apoptosis and late apoptosis percentage increased.The mRNA and protein expressions of Bax,Cyt-c,Caspase 9,Caspase 3 significantly increased(P<0.05),but the mRNA and protein expression of Bcl-2 significantly decreased(P<0.05).Therefore,in this study,it was found that in a certain concentration range,cinobufacin could up-regulate Bax,Cyt-c,Caspase 9,Caspase 3 gene expression and down-regulate Bcl-2 gene to induce apoptosis of human gastric cancer MGC-803 cells.
作者
潘阳
邵淑丽
冯元
李珊珊
焦凯贺
张伟伟
张珍珠
李铁
PanYang;Shao Shuli;Feng Yuan;Li Shanshan;Jiao Kaihe;Zhang Weiwei;Zhang Zhenzhu;Li Tie(College of Life Science and Agroforestry,Qiqihar University,Qiqihar,161006;Key Laboratory of Heilongjiang Province for Resistance Gene Engineering and Protection of Biodiversity in Cold Areas,Qiqihar University,Qiqihar,161006)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第7期2819-2824,共6页
Genomics and Applied Biology
基金
黑龙江省教育厅基本业务专项重点项目(135109104)
黑龙江省高教强省优势特色学科(LTSW201737)
齐齐哈尔大学研究生创新科研项目(YJSCX2018-ZD14)共同资助
