摘要
目的:观察异鼠李素对人肺癌细胞A549、SPC-A1的抑制作用,研究其对CyclinD1、CDK4表达及mTOR磷酸化水平的影响。方法:不同浓度异鼠李素处理人肺癌A549、SPC-A1细胞,采用CCK-8法检测A549、SPC-A1细胞的存活能力;采用划痕实验检测细胞迁移能力;通过Transwell检测细胞体外侵袭能力;采用Western blot法检测A549、SPC-A1细胞中CyclinD1、CDK4、mTOR、p-mTOR蛋白表达;采用RT-PCR法检测细胞中CyclinD1、CDK4 mRNA表达。结果:不同浓度异鼠李素对A549、SPC-A1的增殖具有明显的抑制作用,且随着浓度的升高,抑制作用更加明显。不同浓度异鼠李素作用于A549、SPC-A1后,肺癌细胞的迁移及侵袭能力均有所减弱,接近SC66及顺铂处理水平。与空白对照组比较,不同浓度的异鼠李素可明显抑制CyclinD1、CDK4蛋白表达,并且在一定程度上降低mTOR的磷酸化水平。同时各实验组CyclinD1、CDK4 mRNA表达水平也显著降低。结论:异鼠李素对肺癌细胞增殖具有抑制作用,同时能抑制细胞迁移及侵袭,其机制可能与抑制mTOR信号通路的磷酸化有关。
Objective:To observe the inhibitory effect of isorhamnetin on human lung cancer cells A549 and SPC-A1,to study its effect on the expression of CyclinD1 and CDK4 and the phosphorylation level of mTOR.Methods:Human lung cancer A549 and SPC-A1 cells were treated with different concentrations of isorhamnetin,the survival viabilities of A549 and SPC-A1 cells were determined by CCK-8 method.The cell migration ability was detected by scratch test.Transwell was used to detect the invasiveness of cells in vitro.Western blot was used to detect the expressions of CyclinD1,CDK4,mTOR and p-mTOR proteins in A549 and SPC-A1 cells.The expressions of CyclinD1 and CDK4 mRNA in cells were detected by RT-PCR.Results:Different concentrations of isorhamnetin significantly inhibited the proliferation of A549 and SPC-A1,and the inhibition effect became more obvious with the increase of concentration.After treating A549 and SPC-A1 with different concentrations of isorhamnetin,the migration and invasion abilities of lung cancer cells were weakened and closed to SC66 and cisplatin treatment levels.Compared with the normal control group,different concentrations of isorhamnetin could significantly inhibit the expressions of CyclinD1 and CDK4 proteins and reduce the phosphorylation level of mTOR to a certain extent.At the same time,the expression levels of CyclinD1 and CDK4 mRNA in all the experimental groups were also significantly decreased.Conclusion:Isorhamnetin can inhibit the proliferation of lung cancer cells,as well as cell migration and invasion.The mechanism may be related to the inhibition of phosphorylation of mTOR signaling pathway.
作者
陈观平
李明乾
应栩华
CHEN Guan-ping;LI Ming-qian;YING Xu-hua(Tongde Hospital of Zhejiang Province/Cancer Institute of Integrated Chinese and Western Medicine,Hangzhou 310012,China)
出处
《中药材》
CAS
北大核心
2021年第2期422-427,共6页
Journal of Chinese Medicinal Materials
基金
浙江省中医药科技计划资助项目(2017 ZB017)
