The Study Of Therapeutic Effect And Mechanism Of Pien Tze Huang On Hepatocellular Carcinoma Mice Based On Proteomics And Metabolomics

ObjectiveChinese formula PIEN TZE HUANG(PZH)has been used worldwide to alleviate inflammation-related diseases and recently,PZH was used for treating hepatocellular carcinoma(HCC)and showed positive clinical effect,however,the anti-tumor mechanism of PZH in HCC still is unclear.MethodsIn this study,the HCC xenograft Balb/c mice were treated orally by different dosages of PZH for two weeks,then the isolated tumor tissues were tested by proteomics using Phospho Explorer antibody microarray contained 584 protein phosphorylation sites.The differentiated proteins were analyzed by using Ingenuity Pathway Analysis(IPA)to decipher the potential molecular mechanism.The related cytokines and cell cycle were examined further verification.Then Elisa kits were applied to measure cytokines and D-series Resolvins.Metabolomics based on LC-TOF/MS was applied to quantify the alternation of endogenous SPMs,and Ingenuity Pathway Analysis(IPA)was applied to generate a network and detect the signal pathway.ResultsThe results showed that PZH could inhibit the tumor weight and volume growth.Serum ELISA test results showed that PZH can regulated the expression of inflammatory cytokines including IL-6,TNF-? and IL-1?.87 up-regulated and 11 down-regulated phosphorylated proteins were identified in PZH treated mice.And 21 signaling pathways were associated with inflammation(mainly including anti-inflammation pathways,such as IL-6 and TNFR1/2 pathways),cancer growth(mainly including p53 and FAK pathways)and cell cycle(mainly including G2/M and G1/S checkpoint regulation pathways).To verify the results of IPA,the tumor cells viability,expression of cytokine and cell cycle arrest were tested.These results revealed that PZH achieved anti-tumor in HCC and the underlying mechanisms was regulation of secreting cytokines associated inflammation and inducing G2/M arrest.The metabonomics test results showed that Pien Tze Huang alleviateed the inflammation of tumor by regulating the disordered metabolism of SPMs,especially 17(S)-HDHA,19(20)-EpDPE and D-series resolvins related to KDR,CD5 and GSK3A in NF-?B signaling pathway and PI3K/AKT signaling pathway.ConclusionsIn summary,our study demonstrated the underlying mechanisms of PZH anti-tumor by regulating the IL-6,TNFR1,2 pathways and G2/M DNA damage checkpoint regulation pathway to suppress inflammation and induce G2/M arrest in HCC.It is one of the anti-inflammatory mechanism that PZH regulated the disordered metabolism of SPMs,especially 17(S)-HDHA,19(20)-EpDPE and D-series resolvins related to KDR,CD5and GSK3A in NF-?B signaling pathway and PI3K/AKT signaling pathway to form a network of anti-inflammatory.The findings provided the potential molecular basis for PZH to act as therapy drug or supplements of chemotherapy drugs for human hepatocellular carcinoma in the future.