Preliminary study on the effect and mechanism of Ganning Decoction on hepatocellular carcinoma
Objective:To investigate the anti-hepatocellular carcinoma(HCC)effects of Ganning Decoction and its primary mechanisms.Methods:A total of 40 SD rats were given Ganning Decoction by gavage for 7 days,and serum samples were collect-ed.CCK8 assay was used to detect the effect of serum concentration of Ganning Decoction on the viability of human hepatoma cell line HepG2 at 5%,10%,20%and 30%,respectively.HepG2 cells were divided into blank group and Ganning Decoction-containing serum group.After 24 h of intervention with 5%serum containing Ganning Decoction,cellular proteins were extracted and processed for DIA quantitative proteomic analysis,including the establishment of atlas database and DIA data collec-tion and analysis.The expression of TAGLN protein was detected by Western blot(WB).10 BALB/c nude mice were divided in-to model group and Ganning Decoction group.The HCC model was established by subcutaneous injection of human liver cancer cell line MHCC-97H.After successful modeling,0.9%normal saline or Ganning Decoction was given by gavage for 14 days.The subcutaneous tumor volume was measured every one week from the beginning of modeling to the end of administration and the drew tumor growth curve.After the end of administration,the tumors were dissected,and the histopathological examination of the tumors were detected by hematoxylin-eosin(HE)staining.Results:Ganning Decoction-containing serum decreased HepG2 cell vi-ability in a dose-dependent manner(P<0.05,P<0.01).5%Ganning Decoction-containing serum was used for subsequent experi-ments.DIA quantitative proteomics analysis showed 36 up-regulated proteins and 23 down-regulated proteins(P<0.05).GO en-richment analysis showed cellular component,molecular function and biological process.KEGG enrichment analysis outcomes pointed out the discrepant proteins were closely related to complement and coagulation cascades,PPAR signaling pathway,etc.Reactome enrichment analysis revealed 20 signaling pathways including exocytosis of platelet alpha granule contents.PPI network showed that 38 proteins such as TGFB1,SERPINE1 and TAGLN were in the network nodes.WB validation results show that the expression of TAGLN in Ganning Decoction group was significantly higher than that in the blank group(P<0.01).The subcu-taneous model of HCC in nude mice showed that Ganning Decoction could inhibit tumors growth(P<0.05),and HE staining showed that Ganning Decoction could promote tumor necrosis.Conclusion:Ganning Decoction can inhibit the proliferation of HepG2 cells and tumors,and promote the tumor necrosis.The mechanism involves multiple biological processes and signaling pathways,which may be related to the regulation of TAGLN protein expression.
Ganning decoctionHepatocellular carcinomaProteomicsTAGLN protein expression
万方数据
维普
