Screening and validation for prognostic biomarker and therapeutic target in pancreatic adenocarcinoma
Objective Identify a novel biomarker for prognosticating the survival of patients and therapeutic target in pancreatic adenocarcinoma(PAAD).Methods The prognostic potential of family with sequence similarity 111 member B(FAM111B)was evaluated using GEPIA2 and Kaplan-Meier plotter databases.According to the median of FAM111B expression level,the datasets from Gene Expression Omnibus(GEO)and RNA sequencing data from The Cancer Genome Atlas(TCGA)were divided into high expression group and low expression group,and the datasets were used to further assessed the prognostic value and analyzed the correlation between FAM111B expression and clinical characterization of PAAD patients.Gene set enrichment analysis(GSEA)was used to explore the role of FAM111B in related pathways.Subsequently,RT-qPCR and CCK-8 assays were performed to identify the functions of silencing FAM111B gene group si-FAM111B on tumor proliferation in PAAD cells in comparison with control group si-NC.Flow cytometry was applied to detect cell cycle of si-NC group and si-FAM111B group.Results Based on TCGA database analysis,FAM111B was found to be higher in PAAD tissues than that in matched normal tissues;the difference was statistically significant(P<0.05).GEO datasets of GSE62452 and GSE183795 were further demonstrated that FAM111B expression was higher in PAAD tissues than that in normal tissues;the differences were statistically significant(t=4.617,6.362;all P<0.05).The clinical tumor grade and stage of PAAD was significantly correlated with the FAM111B expression level.And poor prognosis was significantly correlated with overexpression of FAM111B in PAAD patients;the differences were statistically significant(all P<0.05).GSEA results indicated that FAM111B might be involved in PAAD progression through cell cycle signaling pathway.Furthermore,silencing of FAM111B downregulated the mRNA expression of cell cycle related markers,including cyclin B(CCNB),cyclin C(CCNC),cyclin E1(CCNE),cell division cycle 25A(CDC25A),cell division cycle 25C(CDC25C),cyclin dependent kinase 1(CDK1),cyclin dependent kinase 6(CDK6);the differences between si-NC group and si-FAM111B group were statistically significant(t=13.370,6.769,3.011,6.304,5.441,6.058,11.420;all P<0.05).CCK-8 assay indicated that cell proliferation was significantly suppressed in the si-FAM111B group compared to the si-NC group,and the difference was statistically significant(t=43.710,P<0.05).Flow cytometry showed that the proportion of PANC-1 cells in G2/M phase significantly increased in si-FAM111B group after FAM111B knockdown than that in si-NC group(t=8.179,P<0.01).Conclusion FAM111B had the potential to be an independent biomarker of poor prognosis in PAAD patients and the therapeutic target for treating PAAD.
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