Abstract:
Objective: To study the target of emodin in the treatment of pancreatic cancer by bioinformatics and cell experiments.
Methods: The pancreatic cancer transcriptome datasets GSE62452 and GSE28753 were obtained from the Gene Expression Omnibus(GEO) database, and the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and PharmMapper Server were used to obtain the target of emodin. The target genes of the two were cross-screened to obtain the potential target genes of emodin in pancreatic cancer. The DAVID database was used for KEGG and GO functional enrichment analysis of target genes, and GEPIA database was used for differential expression and survival analysis of potential target genes. Pancreatic cancer cells were cultured
in vitro and cell counting kit-8(CCK-8) assay was used to detect the effect of different concentrations of emodin on the proliferation of pancreatic cancer cells. The cells were divided into normal control group and different concentrations of emodin group. The real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the mRNA expression of plasminogen activator urokinase(
PLAU), tyrosine kinase receptor(
MET), epoxide hydrolase 2(
EPHX2), and matrix metalloproteinase 1(
MMP1). Western blotting was used to detect the expression of EPHX2 protein.
Results: Seventeen common target genes of emodin and pancreatic cancer were obtained. Functional enrichment analysis showed that the target genes were mainly enriched in extracellular space, extracellular vesicle, serine endopeptidase activity, protein hydrolysis, calcium ion binding and other pathways. Differential expression analysis showed that there were significant differences in
MMP1, EPHX2, PLAU, MET, and
TTR gene expression between pancreatic cancer tissues and normal tissues(
P<0.05). Survival analysis results showed that
MMP1, EPHX2, PLAU, MET, TTR genes had statistically significant differences between the low expression group and the high expression group in pancreatic cancer(
P<0.05). The results of in vitro experiments showed that the survival rate of pancreatic cancer cells was decreased with the increase of emodin concentration(
P<0.05). Compared with the normal control group, the mRNA expression levels of
PLAU, MMP1, MET, EPHX2 genes in the 20 umol/LEmo group and 40 umol/LEmo group were increased, and the expression level of EPHX2 protein was increased(all
P<0.05).
Conclusion: EPHX2 may be the target of emodin in pancreatic cancer.