Clinical Significance Of ZFX Expression In Intrahepatic Cholangiocarcinoma And Its Relationship With Biological Function

Objective:To investigate the expression of ZFX in intrahepatic cholangiocarcinoma and its relationship with clinicopathological features,and to explore its effect on the prognosis of intrahepatic cholangiocarcinoma.In vitro si RNA silenced ZFX gene in human intrahepatic cholangiocarcinoma HCCC-9810 cells,and observed the effect on apoptosis and migration of HCCC-9810 cells,in order to provide new molecular therapy for intrahepatic cholangiocarcinoma.Methods:In the first part,the tissue wax blocks of patients with intrahepatic cholangiocarcinoma who underwent surgery were retrospectively collected in the department of Hepatobiliary and Pancreatic Surgery of the first Affiliated Hospital of Anhui Medical University from February 2012 to February 2017.The clinical records of all patients were complete.A total of 60 intrahepatic cholangiocarcinoma tumor tissues,50 adjacent normal bile duct tissues,and 15 intrahepatic bile duct stone tissues were selected.The cancer tissue was used as the experimental group,the adjacent cancer group and the stone group were used as the control group,and the expression of ZFX in each group was detected by immunohistochemistry(PV-9001two-step method).To explore the relationship between the expression of ZFX in cancer tissues(high expression and low expression)and clinicopathological features.The Kaplan-Meier method was used to evaluate the overall survival curve of the two groups,and the Cox regression analysis model was used to determine the independent influencing factors of prognosis in patients with ICC.In the second part,human intrahepatic cholangiocarcinoma HCCC-9810 cells were cultured in vitro,and cells were transfected with si RNA(ZFX-si RNA)containing ZFX target gene and si RNA(NC-si RNA)with nonsense sequence,respectively.The experiment was divided into three groups: ZFX-si RNA group,NC-si RNA group and blank control group.After 72 hours of cell culture,the changes of ZFX gene protein levels in cells were detected by Western blot.Finally,cell apoptosis was detected by flow cytometry,and cell migration ability was examined by cell scratch test.Results:1.The high expression rate of ZFX protein in intrahepatic cholangiocarcinoma,paracancerous tissues and intrahepatic bile duct stones was73.3%,38% and 26.7%,respectively,and the high expression rate in tumor tissues was significantly higher than that in adjacent tissues(?2=13.914,P < 0.001)and normal intrahepatic bile duct stones(?2=11.343,P=0.001).2.In patients with intrahepatic cholangiocarcinoma,ZFX has a high expression rate of patients with elevated serum CEA,low tumor differentiation,lymph node metastasis,and high TNM stage,and the expression of serum CEA and tumor differentiation is statistically significant.(P<0.05).3.Kaplan-Meier analysis showed that the overall survival rate of patients with high ZFX expression was significantly lower than that of patients with low ZFX expression(P=0.006).Multivariate Cox regression analysis showed that ZFX expression was an independent influencing factor for poor prognosis in patients with intrahepatic cholangiocarcinoma(HR=2.635,95% CI=1.044-6.651,P=0.040).4.In vitro,ZFX-si RNA was transfected into HCCC-9810 cells for 72 h.Compared with NC-si RNA group(control group)and Cell(blank control),western blot analysis showed that the expression of ZFX protein in HCCC-9810 cells was down-regulated in the ZFX-si RNA group.5.Apoptosis experiments showed that the apoptosis rate of cells in ZFX-si RNA group was higher than that in Cell group(t=22.623,P<0.05)and NC-si RNA group(t=24.854,P<0.05).6.The cell scratch test showed that the cell migration number in the Cell group and the NC-si RNA group was significantly higher than that in the ZFX-si RNA group(P<0.05),but there was no significant difference between the Cell group and the NC-si RNA group.Concluision:1.ZFX is highly expressed in intrahepatic cholangiocarcinoma.The expression of ZFX is related to the malignant clinicopathological features of intrahepatic cholangiocarcinoma.High expression of ZFX is an independent factor for poor prognosis of intrahepatic cholangiocarcinoma.2.Silencing the expression of ZFX gene promotes apoptosis and inhibits cell migration of intrahepatic cholangiocarcinoma HCCC-9810 cells.