摘要
目的实验以非小细胞肺癌H1299细胞(简称H1299细胞)为研究对象,主要探讨白藜芦醇联合野生型p53质粒对H1299细胞凋亡作用的影响以及进一步探讨其作用机制。方法在H1299细胞中导入野生型p53质粒,加入不同浓度的白藜芦醇,通过MTT法确定白藜芦醇联合野生型p53质粒的最佳作用浓度,并检测白藜芦醇联合野生型p53质粒对细胞增殖的影响;光镜下观察细胞形态学的变化;流式细胞术检测白藜芦醇联合野生型p53质粒对H1299细胞凋亡率的影响;RT-PCR实验方法检测细胞凋亡相关基因Bcl-2、Bax、Caspase3、Caspase9的表达水平。结果白藜芦醇联合野生型p53质粒能明显抑制H1299细胞的增殖,诱导H1299细胞凋亡,明显抑制Bcl-2基因的表达水平,促进Bax、caspase3、caspase9基因的表达水平。结论白藜芦醇联合野生型p53质粒能够明显抑制H1299细胞的增殖,可能通过促凋亡基因Bax、凋亡蛋白酶Caspase3,Cas⁃pase9的表达,抑制抑凋亡基因Bcl-2的表达,从而激活细胞中线粒体介导的凋亡通路,诱导H1299细胞凋亡。
Objective To investigate the effect of resveratrol combined with wild-type p53 plasmid on the apoptosis of H1299 cells and its mechanism.Methods Wild-type p53 plasmid was introduced into H1299 cells,and different concentrations of resveratrol were added.The optimal concentration of resveratrol combined with wild-type p53 plasmid was determined by MTT method.The effect of resveratrol combined with wild-type p53 plasmid on cell proliferation was detected,and the morphological changes of cells were observed under light microscope.The effect of resveratrol combined with wild-type p53 plasmid on the apoptosis rate of H1299 cells was detected by flow cytometry RT-PCR was used to detect the expression of apoptosis related genes Bcl-2,Bax,Caspase3 and caspase9.Results The results showed that resveratrol combined with wild-type p53 plasmid could significantly inhibit the proliferation of H1299 cells,induce apoptosis of H1299 cells,significantly inhibit the expression level of bcl-2 gene,and promote the expression level of Bax,Caspase3 and caspase9 gene.Conclusions Resveratrol combined with wild-type p53 plasmid can significantly inhibit the proliferation of H1299 cells,possibly through the expression of Pro apoptotic genes Bax,Caspase3 and caspase9,and inhibit the expression of anti apoptotic gene Bcl-2,thereby activating the mitochondrial mediated apoptosis pathway and inducing apoptosis of H1299 cells.
作者
张平
杜爱萍
黄霞丽
Zhang Ping;Du Aiping;Huang Xiali(Xiangyang Polytechnic,Xiangyang Hubei 441050,China)
出处
《襄阳职业技术学院学报》
2021年第2期89-92,107,共5页
Journal of Xiangyang Polytechnic
基金
2018年度湖北省教育厅科学研究计划指导性项目“p53与Si-MDM2共表达质粒对H1299细胞系凋亡作用的影响及其机制的探讨”(B2018466)。
