Changes In PLK1 And Aurora A Levels In Tumor Tissue And Serum Of Patients With Extrahepatic Cholangiocarcinoma And Their Clinical Significance

Cholangiocarcinoma（CCA）is a highly malignant tumor that arises from intrahepatic or extrahepatic bile duct mucosa epithelial cells,extrahepatic bile duct carcinoma is the most common type of cholangiocarcinomas in the clinical.The global epidemiological investigation showed that the morbidity and mortality of extrahepatic bile duct carcinoma have increased in recent 20 years,but there is no relevant etiological explanation in the world.There are many factors,multi-stage,multi-step and multiple genes in the process of tumors development,however,the abnormal regulation of mitosis and the imbalance of cell proliferation are the most important mechanism of tumorigenesis.Polo-like kinase 1（polo-like kinase 1,PLK1）and Aurora A are important regulatory enzymes which involved in the initiation and maintenance of cell mitosis,and studies have found that PLK1 and Aurora A may control the initiation of mitosis by Aurora A-Bora-PLK1.At present,PLK1 and Aurora A are reported in many tumors of the digestive system,such as esophageal cancer,gastric cancer,colorectal cancer,liver cancer,pancreatic cancer and so on,however,they have not been reported in extrahepatic bile duct carcinoma,and the combination of less tissue and serum to study the relationship between the expression of PLK1 and Aurora and the development of tumor in extrahepatic bile duct carcinoma.Objective:In the topic,the EliVision^TMPlus immunohistochemical technique（immunohistochemistry,IHC）and ELISA（double antibody sandwich method）were used to detect the expression level of PLK1 and Aurora A in the tissues and serum of patients with extrahepatic bile duct carcinoma,and to discuss the following questions:1.To investigate the expression of PLK1、Aurora A in extrahepatic bile duct carcinoma,cancer adjacent normal tissues,and its relationship with clinical pathological features(Clinical stage,differentiation,tumor site,nerve invasion and lymph node metastasis;2.To investigate the relationship between the expression of PLK1 and Aurora A in extrahepatic bile duct carcinoma tissue;3.To investigate the expression of PLK1、Aurora A in serum of preoperative and postoperative patients with extrahepatic bile duct carcinoma and healthy person;4.To investigate the consistency of the expression of PLK1、Aurora A using immunhistochemisty and enzyme linked immunosorbent assay.Methods:Collected 54 cases of extrahepatic bile duct carcinoma tissues and 20 cases of cancer adjacent normal tissues of the 251 st hospital of PLA from February 2009 to September 2015,serum specimens from January 2013 to December 2015 in the 251 st hospital of PLA,there are 25 cases of extrahepatic bile duct carcinoma in patients with preoperative and postoperative serum 15 days,15 healthy volunteers.The expression of PLK1 and Aurora A in extrahepatic bile duct carcinoma tissue and serum were respectively detected;To analyze the relationship between PLK1 and Aurora A and sex,age,tumor differentiation,perineural invasion,TNM stage,location and lymph node metastasis,and the relationship between them;Two methods were used to detect the consistency of the expression of PLK1 and Aurora A.The experimental data results were analyzed by SPSS19.0 statistical software.Results:1.The positive expression of PLK1 and Aurora A are mainly in the nucleus,cytoplasm is also expressed.In patients with extrahepatic bile duct carcinoma,carcinoma tissues PLK1 and Aurora A positive expression rate were significantly higher than that of bile duct tissue adjacent to carcinoma（66.7% vs.25.0%;63.0% vs.66.7%,P < 0.05）.2.The expression of PLK1 and Aurora A are not related with sex,age,neurological involvement and location of tumors in extrahepatic bile duct carcinoma tissue（P>0.05）,and they are related with the degree of tumor differentiation,TNM stages and lymph node metastasis（P < 0.05）,The expressions of PLK1 and Aurora A in poorly differentiated,lymph node metastasis and TNM stage III/IV of extrahepatic bile duct carcinoma are significantly higher than those of highly and moderately differentiated,no lymph node metastasis and TNM stage I/II Bile duct carcinoma.3.The preoperative serum concentrations of PLK1 and Aurora A are significantly higher than those of healthy people（434.85 pg/mL vs.256.00 pg/mL;644.64 pg/mL vs.375.73 pg/mL,both P<0.05）,and The postoperative serum concentration of PLK1 and Aurora A are significantly lower than that before operation in patients with extrahepatic bile duct carcinoma（P<0.05）.4.Immune histochemical method is used to inspect PLK1 in 25 cases of extrahepatic bile duct carcinoma tissues of 17 cases of positive expression,8 cases were negative expression;Aurora A in 25 cases of extrahepatic bile duct carcinoma tissue,16 cases of positive expression in 9 cases of negative expression;Coupled with ELISA test results,the Kendall test analysis,The expression level of PLK1 in cancer tissues is consistent with the concentration of serum（r=0.55,P=0.55）,The expression level of Aurora A in cancer tissues is also consistent with the concentration of serum（r=0.64,P<0.01）.5.To compare the expression of PLK1 and Aurora A in 54 cases of extrahepatic bile duct carcinoma,the Spearman rank correlation analysis,a positive correlation between the expression of PLK1 and Aurora A in extrahepatic bile duct carcinoma tissue（r=0.47,P<0.01）;To compare the level of PLK1 and Aurora A in Preoperative serum of 25 patients with extrahepatic bile duct carcinoma,the Spearman rank correlation analysis,the expression of PLK1 and Aurora A in serum of patients was positively correlated（r=0.71,P<0.01）.Conclusion:1.PLK1 and Aurora A are highly expressed in the tissues and serum of patients with extrahepatic bile duct carcinoma,and low expression in the normal tissues adjacent to the tumor and healthy people,this may be an important biological feature of extrahepatic bile duct carcinoma.2.PLK1 and Aurora A are correlated with the degree of differentiation,lymph node metastasis and TNM stage of extrahepatic bile duct carcinoma,that it may be involved in the invasion and metastasis of cholangiocarcinoma,can provide the basis for clinical in extrahepatic biliary carcinoma diagnosis and treatment.3.PLK1 and Aurora A are consistent in the two methods of immunohistochemistry and ELISA,and ELISA is easier to be popularized in clinical practice,which can provide a more accurate basis for the judgment of extrahepatic cholangiocarcinoma and the monitoring and treatment of disease.4.There was a positive correlation between PLK1 and Aurora A in extrahepatic bile duct carcinoma tissue and serum,the combined detection can both gene detection sensitivity and specificity,and for clinical screening of extrahepatic bile duct carcinoma,and provide a theoretical basis for the research and development of anticancer drugs and prognosis.