Fluorescence in situ hybridization technology to improve cholangiocarcinoma clinical diagnostics and related mechanisms

Background cholangiocarcinoma originated in the biliary epithelial cell malignancy, accounting for about 15% of the malignant tumors of the hepatobiliary system. Sectional imaging studies are often difficult to find a space-occupying lesions of the bile duct, cholangiocarcinoma cytology in the preoperative diagnosis is very important. Special anatomical location makes the biliary tract specimens get difficult, but a small amount of specimens made, resulting in the cytological diagnosis low. Therefore, need to find a more sensitive and reliable method to improve the diagnosis rate of cholangiocarcinoma. Widely present in the tumor tissue chromosome aneuploidy changed. Detected by fluorescence in situ hybridization (Fluorescencein situ hybridization, FISH) chromosome abnormality can assist cancer diagnosis, but cholangiocarcinoma applications rarely reported. On the other hand, Aurora kinase A (AURKA) cells have important regulatory proteins during mitosis, the expression levels can cause uneven chromosome segregation, cell chromosome aneuploidy change. It has now been confirmed AURKA over-expression in many human tumors, and the biological behavior of the tumor. But there is still a lack of understanding of cholangiocarcinoma in AURKA expression. 2) whether the application of fluorescence in situ hybridization techniques to improve clinical bile duct research purposes, we hope that through the biliary tract cancer cell lines, cells of pathological and clinical biliary brush specimens clear: 1) cholangiocarcinoma cells to chromosome aneuploidy changed; The tumor diagnosis rate of cytology specimens; 3) AURKA expression levels in cholangiocarcinoma; 4) whether the biological behavior of the of AURKA expression of cholangiocarcinoma correlation. Research methods we choose to have a wide range of tumor cytology research base and the FDA certification UroVysion mixed probe, respectively 1) QBC939 (cholangiocarcinoma cell lines), GBC-SD (gallbladder cancer cell line), CC-HEL-1 (fetal liver cell line) in situ hybridization, compare biliary tumor cells and normal cells of chromosome hybridization signal number; 2) of cholangiocarcinoma and inflammatory bile duct tissue in situ hybridization, the hybridization signal number compare two kinds of organizations; 3) of 27 patients with biliary cell brush specimens in situ hybridization, pathology or clinical follow-up results as the gold standard, FISH and conventional cytology value for tumor diagnosis. Real-Time PCR and Western Blot method detection QBC939 GBC-SD, CC-HEL-1 cell lines and tissues isolated biliary epithelial cells and hepatocytes AURKA gene expression, comparing different cells AURKA mRNA and protein level differences . Detected by immunohistochemistry method in 29 cases of cholangiocarcinoma of AURKA the expression, and inflammatory biliary tissue contrast, analysis of the of AURKA expression level and tumor pathological stage and histological grading correlation. The results of fluorescence in situ hybridization showed: the biliary tumor cell lines and cholangiocarcinoma memory change in a wide range of cell chromosome aneuploidy, chromosome disomic substantially normal tissue cells. Application the UroVysion probe by FISH technique detected 27 cases of bile duct cell brush specimens No. 3, No. 7, 17 and 9p21 gene locus, positive abnormal number of chromosomes criteria (ie: multiple chromosomal polysomy cells ≥ 5 trisomy cells ≥ 10) or a single chromosome, biliary tumor diagnostic sensitivity, specificity, positive predictive value and negative predictive value: 50%, 100%, 100%, 31.3%, respectively; conventional cytology The diagnostic sensitivity, specificity, positive predictive value, and negative predictive values ??were: 22.7%, 100%, 100% and 22.7%, between the two, did not reach statistical significance (P = 0.146). In the present study, the chromosome 3 polysomy FISH-positive patients with chromosomal abnormalities main form, and can be seen 9p21 homozygous deletion. AURKA mRNA and protein levels were detected using Real-Time PCR and Western Blot method, found that fetal liver cells and normal biliary epithelial and liver cells: AURKA expression levels were significantly higher in tumor cell lines QBC939 GBC-SD; immune staining AURKA increased expression in cholangiocarcinoma and AURKA expression level of correlation with tumor staging and histological type. The widespread conclusion cholangiocarcinoma cell aneuploidy change. Improve tumor diagnosis rate of biliary cytology brushes can to a certain extent by fluorescence in situ hybridization; AURKA expression levels were significantly higher in cholangiocarcinoma, and its expression levels associated cholangiocarcinoma biological behavior. Speculated AURKA may be involved in the occurrence of cholangiocarcinoma by causing chromosome aneuploidy change.