Expression And Significance Of USP4 In Cholangiocarcinoma

Objective This study focused on the expression of ubiquitin-specific protease 4(USP4)in cholangiopathy tissues,the expression levels of USP4 measured at both protein and RNA levels,and analyzed the clinical data of patients,and analyzed the possible correlation between the experimental data of bile duct carcinoma group(T),bile duct benign lesion(B)and distal margin negative normal bile duct tissue(N)and whether there were statistical differences between the two groups,predicting whether USP4 can be used as a molecular marker for the diagnosis of cholangiocarcinoma.Methods Twelve patients with cholangiocarcinoma,benign lesion and negative distal margin of malignant tissue were collected from our hospital between September 2018 and December 2020,aged 48-78 years old.All subjects met the diagnosis and inclusion criteria,signed the consent form informed and voluntarily,and clinical data of the patients were collected.Using hematoxylin-eosin staining(HE),immunohistochemical staining(IHC),Western blot and polymerase chain reaction(RT-PCR),we detected the expression of USP4 in three fresh experimental tissues from two aspects of protein and m-RNA levels.The experimental data between the groups were compared to compare whether there was difference between the groups.Meanwhile,the clinical data of patients with cholangiocarcinoma were collected to analyze whether the expression level of USP4 was correlated with the clinical data.The results of experiment used SPSS 25.0 statistical software to analyze the experimental results.The measurement data were expressed as mean ± standard deviation(X ±S),the counting data were counted by Fisher accurate probability method,and the comparison between groups adopts t test.P<0.05 difference was statistically significant.Results 1.The results of HE experiments showed that the differences of three kinds of tissues were observed by comparing cholangiocarcinoma,benign bile duct lesions and normal bile duct tissues from cell size,morphology,heterogeneity,nucleus morphology,mitotic image;2.The IHC results showed that the color depth and positive color area of USP4 protein in the three tissues were as follows: cancer tissue(T)> benign lesion(B)> normal tissue(N);3.The results of western blot showed that USP4 protein expression in three tissues showed that cancer tissue(T)>benign lesion(B)and normal tissue(N)were).The P values <0.0001,which showed that there were statistical differences between the two groups.It has obvious statistical significance;4.RT-PCR results showed that the m-RNA expression of USP4 in three tissues was: cancer tissue(T)> benign lesion(B)and normal tissue(N).The P <0.0001,which showed that there were significant statistical differences between the two groups.It has obvious statistical significance;5.The ROC curvature: m-RNA in cancer tissue(T)/ benign lesion(B)and the results of ROC curve analysis showed that: the diagnosis of cancer tissue(T),benign lesions(B)had differential significance;6.The expression of USP4 in cancer tissues and the data of the patients were analyzed.The expression level of USP4 protein was correlated with the gender of the patients and the clinical characteristics of the preoperative CEA(P(27)0.05),but not with other clinicopathological characteristics(P>0.05).Conclusion The up-regulated expression of USP4 in cholangiocarcinoma tissue can be used as a molecular marker for differential diagnosis of benign and malignant cholangiocarcinoma.