| ObjectiveTo investigate the antitumor effect of Polygonatum odoratum Druce alcoholic extract on gastric cancer cells and to further explore its mechanism.Methods1.The alcohol extraction of Polygonatum odoratum Druce: We used alcohol extraction method to separate and extract 70% ethanol at 60 ℃ for50 min.The alcohol extract concentration of Polygonatum odoratum was 0.824mg/L.2.Cell culture and drug concentration screening: Gastric cancer cell lines BGC-823 and SGC-7901 were cultured in vitro.The drug concentration of the alcohol extract was screened and the appropriate drug concentration was selected for the next experiment.3.The main components of Polygonatum odoratum Druce alcohol extract were verified by HPLC.4.CCK-8 assay was used to discover the proliferation of gastric cancer cells.5.Clonal formation assay was used to investigate clonal proliferation of gastric cancer cells.6.Cell scratch experiments was used for detecting migration ability of gastric cancer cells.7.Transwell assay was used to explore invasion and migration of gastric cancer cells.8,Western blotting experiments was used to detect the expression of proliferation-related proteins(Ki67,MCM5),epithelial I-mesenchymal transition(EMT)-related proteins expression(E-cadherin,N-cadherin,ZO-1,Snail and TWIST)and WEE1 protein expression and phosphorylation status of CDK1-T14 and CDK1-Y15 in WEE1 signaling pathway.Results1.The main components of alcohol extract of Polygonatum odoratum Druce were verified by HPLC.2.According to the screening of the alcohol extract concentration of Polygonatum odoratum Druce,the drug concentrations of 0 μg/m L,20 μg/m L,40 μg/m L and 80 μg/m L were selected for the next experiment.3.CCK-8 assay showed the alcohol extract inhibited the proliferation of gastric cancer cell lines BGC-823 and SGC-7901(p<0.05).4.Cloning and formation experiments showed that the alcohol extract inhibited the proliferation of gastric cancer cell lines BGC-823 and SGC-7901,and the number of clones decreased(p<0.01).5.The cell scratch experiment showed that the alcohol extract inhibited the cell migration of gastric cancer cell lines BGC-823 and SGC-7901,the cell scratch distance was relatively increased,and the cell migration rate was decreased(p<0.01).6.Transwell assay showed that the alcohol extract of Polygonatum odoratum Druce inhibited the invasion and migration of gastric cancer cell lines BGC-823 and SGC-7901,and the number of invaded and metastatic cells decreased(p<0.01).7.The results of western blot showed that compared with the sham group,the expression of proliferation-related protein Ki67 and MCM in Polygonatum odoratum alcohol extract group decreased(p<0.05)and was concentration-dependent.Compared with the control group,the expression of EMT-related proteins E-cadherin and ZO-1 increased,while the expression of N-cadherin,Snail and TWIST decreased(p<0.05),which was concentration-dependent.WEE1 protein expression and CDK1-T14 and CDK1-Y15 protein expression decreased(p<0.01).ConclusionsIn this study,the results showed that the alcoholic extracts of Polygonatum odoratum Druce can significantly inhibit the proliferation,invasion and metastasis of gastric cancer cells,and the mechanism of its anti-tumor effect may be related to the inhibition of EMT of gastric cancer cell lines by the alcoholic extracts of Polygonatum odoratum Druce,which provides experimental basis for Polygonatum odoratum Druce as an anticancer drug of traditional Chinese medicine. |
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