Objectives:Ziyin Huatan Recipe(ZYHT),a traditional Chinese medicine comprised of Lilii Bulbus,Pinelliae Rhizoma,and Hedyotis Diffusa,has shown promise in treating gastric cancer(GC).However,its potential mechanism has not yet been clearly addressed.This study aimed to predict targets and molecular mech-anisms of ZYHT in treating GC by network pharmacology analysis and to explore the role of ZYHT in GC both in vitro and in vivo.Methods:Targets and molecular mechanisms of ZYHT were predicted via network pharmacology analy-sis.The effects of ZYHT on the expression of metastasis-associated targets were further validated by Western blot and quantitative real-time polymerase chain reaction.To explore the specific molecular mechanisms of the effects of ZYHT on migration and invasion,the runt-related transcription factor 3(RUNX3)gene was knocked out by clustered regularly interspaced short palindromic repeats/Cas9,and lentiviral vectors were transfected into SGC-7901 cells.Then lung metastasis model of GC in nude mice was established to explore the anti-metastasis effect of ZYHT.Western blot and immunohistochemistry were used to explore the impact of ZYHT on the expression of metastasis-related proteins with or without RUNX3 gene.Results:The network pharmacology analysis showed that ZYHT might inhibit focal adhesion,migration,invasion and metastasis of GC.ZYHT inhibited the proliferation,migration and invasion of GC cells in vitro via regulating the expression of metastasis-associated targets.Knocking out RUNX3 almost completely reversed the cell phenotypes(migration and invasion)and protein expression levels elicited by ZYHT.In vivo studies showed that ZYHT inhibited the metastasis of GC cells to the lung and prolonged the sur-vival time of the nude mice.Knocking out RUNX3 partly reversed the metastasis of GC cells to the lung and the protein expression levels elicited by ZYHT.Conclusion:ZYHT can effectively inhibit the invasion and migration of GC in vitro and in vivo,and its molecular mechanism may relate to the upregulation of RUNX3 expression.

Department of Traditional Chinese Medicine,Changzheng Hospital,Naval Medical University,Shanghai 200003,China;Strategic Support Force Xingcheng Special Duty Sanatorium,Xingcheng 125100,Liaoning Province,China;Cancer Institute,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China

[1]Shang-jin Song;Xuan Liu;Qing Ji;Da-zhi Sun;Li-juan Xiu;Jing-yu Xu;Xiao-qiang Yue-.Ziyin Huatan Recipe,a Chinese herbal compound,inhibits migration and invasion of gastric cancer by upregulating RUNX3 expression)[J].结合医学学报（英文版）,2022(04):355-364

Ziyin,Huatan,ZYHT,Lilii,Bulbus,Pinelliae,Diffusa

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