Effect And Mechanism Of Berberine On Metastasis Of Nasopharyngeal Carcinoma Cells

[Background and Objective] Nasopharyngeal carcinoma(NPC)is a malignant tumor with high incidence and high mortality in coastal and central provinces of China.It is of great significance to develop new treatment methods and subsequently improve the treatment of NPC.Berberine,an extract of Coptis chinensis,is effective on suppressing both solid tumors and hematological tumors,including NPC.However,the role and mechanism of Berberine in nasopharyngeal carcinoma are still inadequate.Using the second generation high-throughput sequencing technology,the effect of berberine on the overall transcriptome of NPC cell line S18 was analyzed in this study,and the regulation and biological function of berberine on autophagy of S18 cells were studied,which further replenished the molecular mechanism of berberine inhibiting NPC.[Methods] The effects of berberine on the growth,proliferation,migration and invasion of S18 cells were analyzed by CCK8,plate clone formation,Transwell migration and invasion assay.Using the second generation high-throughput transcriptome sequencing technique,the effects of berberine treatment on the expression profiles of S18 protein coding mRNA and long-chain non-coding RNA were detected,and the correlation was analyzed by GO and signal pathway enrichment.By means of qPCR,western blot,RNA interference and autophagy inhibitor treatment,the regulatory effects of berberine on autophagy-related gene expression and protein level,autophagy level,autophagy regulatory pathway activity and autophagy function were detected.[Results] CCK8 results showed that berberine inhibited the growth of S18 cells in a concentration-dependent manner.The results of plate colony formation showed that berberine treatment could significantly inhibit the size and number of clone of S18 cells.The results of Transwell migration and invasion experiments showed that berberine treatment could significantly inhibit the number of S18 cells that degraded Matrigel,through porous membrane.Differential expression genes inducing by berberine were significantly enriched in GO processes such as misfolded protein endoplasmic reticulum stress,DNA replication,apoptosis and inflammation.Signal pathway analysis showed that differential genes were significantly enriched in tumorigenesis and autophagy regulatory pathways such as MAPK,Hippo,PI3K/AKT,TGF-β and p53.qPCR,Western blot and other results showed that berberine treatment significantly up-regulated the expression and protein level of autophagy-related genes such as ULK1,and MAP1LC3 B,up-regulated the expression of SQSTM1,down-regulated the protein level of SQSTM1,and promoted the accumulation of LC3B-II.At the same time,berberine could significantly inhibit the activity of AKT/mTOR pathway.When the autophagy was inhibited by ATG5 knockdown,the inhibitory effect of berberine on S18 was significantly antagonized.[Conclusion] Berberine treatment can significantly inhibit the growth,proliferation,migration and invasion of S18 cells,and berberine treatment can significantly change the transcriptome profile of S18 cells,inhibit tumor-promoting signals such as AKT/mTOR,induce autophagic death and inhibit the growth and malignant characteristics of S18 cells.