Background:Uncontrolled inflammation plays an important role in the initiation and
progression of tumors. The repeated circulation and continuous stimulation of gallbladder epithelium
caused by gallstones is an important risk factor for gallbladder cancer.Methods:To study pathogenesis, samples were collected for chronic cholecystitis caused by
gallstones and early and advanced gallbladder cancer with gallstones and subjected to RNA-seq
analysis. Gene Ontology and Kyoto Gene and Genome Encyclopedia analyses were used to elucidate
the protein–protein interaction network and identify differentially expressed genes.Results:Nine potential molecular markers, VTN, CHAD, AKR1C4, ABCC2, AOX1, ADH1A,
ADH1C, PLA2G2A, and CYP4F3, with elevated expression gradients in cholecystitis and early
and advanced gallbladder cancer, were identified. Using qPCR and immunohistochemistry on
clinical tissues, we confirmed three factors, VTN, CYP4F3, and AOX1, to be worthy of further
research. To demonstrate that these three genes are potential molecular markers for gallbladder
cancer, their cellular biological functions were confirmed in gallbladder cancer cell lines through
siRNA transfection.Conclusion:The potential molecular markers CYP4F3, VTN, and AOX1 for cholecystitis and
different stages of gallbladder cancer were identified. Further studies on differentially expressed
genes vital in gallbladder cancer progression can help provide potential targets for the early diagnosis
and treatment of gallbladder cancer.