Screening Of Traditional Chinese Medicine Compounds With Anti-nasopharyngeal Carcinoma Effect And The Studying Of Mechanisms Of Baicalein On Inhibition Of Proliferation,Invasion And Migration Of Nasopharyngeal Carcinoma Cells

Objective and Contents:In this study,17 kinds of traditional chinese medicine compounds were screened to find out the monomer having the ability to inhibit the proliferation of nasopharyngeal carcinoma cells and to explore its mechanism.The main contents of the study include three parts:1.Screening traditional chinese medicine compounds that can inhibit the proliferation of nasopharyngeal carcinoma cells;2.To study the effects of Baicalein on proliferation,invasion and migration of nasopharyngeal carcinoma cells;3.To study the mechanism of Baicalein in inhibiting proliferation,invasion and migration of nasopharyngeal carcinoma cells.Methods:1.Using a low differentiated nasopharyngeal carcinoma cell line CNE2 as a model.Using CCK-8 to screen the compounds of traditional Chinese medicine that have the ability to inhibit CNE2 proliferation.2.CCK-8 and Clone formation assay were used to detect the effects ofBaicalein on the proliferation of nasopharyngeal carcinoma cells.Flow cytometry was used to detect the effects of Baicalein on cell apoptosis and cell cycle.Fluorescent staining was used to detect the effect of Baicalein on chromatin of cells.Caspase-3 active kit was used to detect the effects of Baicalein on the activity of Caspase-3 protein.Scratch and heterogeneity adhesion assay were preliminary used to detect the effects of Baicalein on invasion and migration.Transwell invasion and migration assays were used to detect the effects of Baicalein on invasion and migration.3.Network pharmacology was used to predicted the target of Baicalein.Western blot was used to detect the effects of Baicalein on the expression of key proteins in MAPK and p53 signaling pathways.qRT-PCR was used to detect the effects of Baicalein on cell cycle related genes and MMPs gene mRNA.Results:1.CCK-8 was used to screen out six kinds of traditional chinese medicine compounds that inhibit the proliferation of low differentiated nasopharyngeal carcinoma cells from 17 kinds of traditional chinese medicine compounds.The six kinds of traditional chinese medicine compounds were Dihydromyricetin,luteolin,Baicalein,Amentoflavone,Galangin and Theaflavin.2.CCK-8 and Clone formation assay found that Baicalein can significantly inhibit the proliferation of nasopharyngeal carcinoma cells.Flow cytometry found that the apoptosis was significantly increased,and the cell cycle was arrested at S stage.The typical apoptotic features were detected by fluorescence microscopy.And the Caspase-3 activity can be significantly increased by Baicalein.Heterogeneous adhesion assay and Transwell invasion assay found that Baicalein could significantly inhibit the adhesion and invasion ofnasopharyngeal carcinoma cells.Scratch assay and Transwell migration assay found that Baicalein can significantly inhibit the migration of nasopharyngeal carcinoma cells.3.79 targets of Baicalein were predicted by Network pharmacology,and enrichment analysis showed that Baicalein could be regulated by MAPK signaling pathway.Using Western blot to detect the key proteins of MAPK and p53 signaling pathway,it was found that the phosphorylation of JNK and MAPKAPK2 increased significantly,and the expression of p53,Bax and Caspase-3 increased significantly.Using qRT-PCR to detect the cycle related genes,it was found that p53 and p21 mRNA were significantly up-regulated,and CDK2 mRNA was significantly down-regulated.Also,using qRT-PCR to detect the MMPs genes,it was found that MMP1,MMP2,MMP7 and MMP9 mRNA were significantly down-regulated.Conclusions:1.Dihydromyricetin,luteolin,Baicalein,Amentoflavone,Galangin and Theaflavin can inhibit the proliferation of low differentiated nasopharyngeal carcinoma cells.2.Baicalein can inhibit the proliferation and induce apoptosis of nasopharyngeal carcinoma cells by regulating MAPK and p53 signaling pathway.In addition,Baicalein can inhibit the invasion and migration of nasopharyngeal carcinoma cells by down regulating the expression of MMP1,MMP2,MMP7 and MMP9 mRNA.