Objective

2

To screen the differential proteins of Shenhu Banxia Decoction on subcutaneous transplanted tumor of human gastric cancer in nude mice， and to explore the biological basis mechanism of Shenhu Banxia Decoction on gastric cancer.

Methods

2

A subcutaneous transplant tumor model of BALB/C nude mice with human gastric cancer was establish by using MGC803 cell line. Twenty nude mice were randomly divided into control group （sodium chloride injection）， Shenhu Banxia Decoction group （0.63 g/kg Shenhu Banxia Decoction）， fluorouracil （5-FU） group （0.025 g/kg 5-FU） and Shenhu Banxia Decoction +5-FU group （0.63 g/kg Shenhu Banxia Decoction+0.025 g/kg 5-FU）， with 5 mice in each group. Nude mice in each group were given corresponding intervention. And inhibitory effect of Shenhu Banxia Decoction on the growth of gastric cancer cells was observed. Label-free quantitative proteomics （LFQ） was used to analyze differentially expressed proteins in tumor after intervention. The Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG） and other databases were used to enrich and analyze the proteins and information pathways. Western blot was used to analyze the protein expression of S100 calcium-binding protein A9 （S100A9）， Rho GDP dissociation inhibitor （ARHGDIB）， and serine protease inhibitor E2 （SerpinE2）.

Results

2

①Tumor volume in each treatmert group was smaller than that in the control group （

P

<

0.05）. The tumor volume of Shenhu Banxia Decoction+5-FU group was significantly smaller than that of Shenhu Banxia Decoction group and 5-FU group （

P

<

0.05）. ②The tumor inhibition rates of Shenhu Banxia Decoction group， 5-FU group and Shenhu Banxia Decoction +5-FU group were 21.13%， 32.66% and 38.63%， respectively. Compared with the control group， the differences were statistically significant （

P

<

0.05）. Compared with 5-FU group and Shenhu Banxia Decoction group， tumor inhibition rate of Shenhu Banxia Decoction+5-FU group was obviously higher （

P

<

0.05）. ③Compared with the control group， among the differential proteins identified in Shenhu Banxia Decoction group， two proteins ［carboxypeptidase B2 （CPB2）， SerpinE2］ were up-regulated and five proteins ［S100A9， S100 calcium-binding protein A8 （S100A8）， ARHGDIB， lysozyme C2（Lyz2）， recombinant Integrin β2 （ITGB2）］ were down-regulated （

P

<

0.05）. ④The GO analysis showed that differentially expressed proteins play important roles in angiogenesis， polarization of nuclear transcription factor-κB （NF-κB） cells， neutrophil aggregation and other biological processes. KEGG analysis results showed that the main signaling pathways related to differentially expressed proteins were interleukin 17 （IL-17） signaling pathway， complement pathway， coagulation cascade pathway and so on. ⑤Western blot showed that compared with the control group， the S100A9 and ARHGDIB protein expression levels decreased significantly （

P

<

0.01）， and the SerpinE2 protein expression level increased significantly in the nude mice tumor tissues of the Shenhu Banxia Decoction group （

P

<

0.01）. The results were consistent with those of proteomic analysis results.

Conclusion

2

Shenhu Banxia Decoction can inbibit the growth of gastric cancer cells. The anti-gastric cancer effect of Shenhu Banxia Decoction may be related to the regulation of inflammation-related proteins， migration proteins and fibrinolytic proteins， and it is closely related to IL-17 signaling pathway， complement pathway， coagulation cascade pathway.