The Gene Expression Analysis Of ACE2and FZD1in Gallbladder Cancer Tissue And Their Role In Invasion And Metastasis Of Gallbladder Cancer

Part I The gene expression analysis of ACE2and FZD1in gallbladder cancer tissueObjectiveThe study was performed to investigate the role of ACE2and FZD1in invasion and metastasis of gallbladder adenocarcinoma.MethodsThe total RNA were extracted from fresh tissue samples of15gallbladder adenocarcinoma and15chronic cholecystitis. The mRNA transcription levels of ACE2and FZD1were evaluated by RT-PCR. Statistic analysis was performed afte data collection.ResultsACE2mRNA was significantly lower in gallbladder adenocarcinoma than that in chronic cholecystitis (0.062±0.011vs0.297±0.019, P<0.01),while FZDl mRNA was significantly higher in gallbladder adenocarcinoma than that in chronic cholecystitis (0.939±0.016vs0.222±0.019, P<0.01)ConclusionsFZD1expression was upregulated and ACE2expression was downregulated in gallbladder adenocarcinoma compared to chronic cholecystitis in vitro that was constistent with in vitro study in part one. It was suggested that ACE2and FZD1might be involved in the process of gallbladder epithelial malignant transformation that is related to the invasion and metastasis of gallbladder adenocarcinoma.Part Ⅱ Differential expression of ACE2and FZD1and their clinicopathological significance in benign and malignant lesions of the gallbladderObjectiveTo investigate the differential expression of ACE2and FZD1in benign and malignant lesions of gallbladder and evaluate its clinicopathological significance.MethodsRoutinely paraffin-embedded sections were prepared from specimens of gallbladder adenocarcinoma (n=100), peritumoral tissues (n=46), adenoma (n=30), polypus (n=15) and chronic cholecystitis (n=35). EnVisionTM immunohistochemistry was used for detecting the expression level of ACE2and FZD1.Results(1) The ratio of positive FZD1ession was significantly higher in adenocarcinoma of gallbladder (59.0%) than that in peritumoral tissues (26.1%) and other benign lesions including adenoma (20.0%), polypus (13.3%) and chronic cholecystitis (11.4%)(P<0.01). The ratio of positive ACE2expression was significantly lower in adenocarcinoma (50.1%) than that in peritumoral tissues (80.4%), adenoma (83.3%), polypus (86.7%) and chromic cholecystitis (88.6%)(P<0.05). The benign lesions with positive FZDl and/or negative ACE2expression showed atypical hyperplasia in gallbladder epithelium.(2) The ratio of positive staining for FZD1in cases with well-differentiated adenocarcinoma, small tumor size (diameter<2cm), no lymph node metastasis and no peripheral tissue invasion was significantly lower than that in cases with poor-differentiated adenocarcinoma, larger tumor size (diameter>2cm), lymph node metastasis and peripheral tissue invasion (P<0.5or P<0.01). However, the expression of ACE2demonstrated a negative correlation in these cases (P<0.05or P<0.01).(3) The monovariable Kaplan-Meier survival analysis suggested that the histological type, tumor size, lymph node metastasis, peripheral tissue invasion, ACE2and FZDl expression were significantly associated with the average survival time of patients with the gallbladder adenocarcinoma (P<0.05or P<0.01). Furthermore, the survival time of cases with positive staining for FZD1significantly shorter than those with negative staining (P<0.05) while ACE2showed a negative correlation (P<0.05). Interestingly, the overall survival time of cases with FZD1(-)ACE2(+) was significantly longer than those with FZD1(+)ACE2(-). Multivariable Cox analysis revealed that FZD1positive expression (P=0.042) or ACE2ative expression (P=0.040) was negatively correlated with postoperative survival and positively correlated with mortality.ConclusionsThe expression of ACE2and FZD1in gallbladder adenocarcinoma is an independent marker for evaluation of the disease progression, clinical biological behaviors and prognosis. Patients with the positive ACE2expression had better prognosis while patients with the positive expression of FZD1had worse prognosis.