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Objective: To detect Survivin, VEGF-C, CyclinD1, CDC25A and PCNA expression in cholangiocarcinoma to explore its role in the development of cholangiocarcinoma occurrence and mechanism for the diagnosis and treatment of cholangiocarcinoma looking for new targets. Method 1 experimental specimens taken cholangiocarcinoma, the same individual organizations and their adjacent normal bile duct tissues using immunohistochemical 1.1 peroxidase labeled streptavidin (SP) method in 42 patients with cholangiocarcinoma tissues and 20 cases of normal tissues Survivin, VEGF-C and PCNA protein were detected in combination with the patient's age, gender, and tumor size, location, degree of differentiation, lymph node metastasis and other clinicopathological factors were analyzed. Samples were taken through the pathological diagnosis confirmed. 1.2 by flow cytometry 42 cases of cholangiocarcinoma and 20 cases of normal bile duct tissue CyclinD1, CDC25A protein expression, protein expression with which the mean fluorescence intensity (average channel value) to represent. Samples were taken through the pathological diagnosis confirmed 1.3 by RT-PCR was used to detect 42 cases of intraoperative taken fresh cholangiocarcinoma, 17 cases with individual adjacent tissues and 20 cases of normal bile duct tissues Survivin, VEGF-C , CyclinD1, CDC25A and PCNA mRNA expression, intraoperative specimens were taken through the pathological diagnosis confirmed. The Gel-proanalyzer3.1 software analysis RT-PCR products of optical density (optical density, OD), calculated Survivin, VEGF-C, CyclinD1, CDC25A and PCNA OD values ??within the reference gene (glyceraldehyde-3-phosphate dehydrogenase, GADPH ) OD value of the ratio, as a target gene product relative amount of mRNA. 2 Applications The SAS System for Windows v6.12 software for statistical analysis. Experimental data were expressed as mean ± standard deviation (x ± s) that different groups of experimental data using analysis of variance (ANOVA), further pairwise comparisons using t-test (LSD method); immunohistochemical results using the chi-square test; were to test the level α = 0.05, when P lt; 0.05 when statistically significant. Results 1. Immunohistochemical detection of 42 cases of cholangiocarcinoma and 20 cases of normal bile duct tissues Survivin, VEGF-C and PCNA protein expression in cholangiocarcinoma tissues, normal bile duct tissue expression of Survivin positive rates were 78.57% and 25.00%, cholangiocarcinoma tissue were significantly higher than normal tissue; cholangiocarcinoma, normal bile duct tissue expression of VEGF-C positive rates were 83.33% and 30.00%, bile duct cancer tissue was significantly higher than normal tissue; cholangiocarcinoma, bile duct normal expression of PCNA positive rates were 71.19% and 25.00%, the positive rate of cholangiocarcinoma tissues was significantly higher than normal tissue. (2) by flow cytometry 42 cases of cholangiocarcinoma tissues and 20 cases of normal bile duct tissues CyclinD1 and protein expression of CDC25A cholangiocarcinoma, normal bile duct tissues CyclinD1 protein expression was 530.253 ± 27.823,497.416 ± 38.019, cholangiocarcinoma CyclinD1 tissue protein than normal tissue; cholangiocarcinoma, normal bile duct tissues CDC25A protein expression was 534.029 ± 35.674,497.276 ± 42.343, cholangiocarcinoma tissue CDC25A protein expression than normal tissue. 3 RT-PCR was used to detect 42 cases of cholangiocarcinoma, 17 cases of the same individual adjacent normal bile duct tissues and 20 cases of bile duct tissues Survivin, VEGF-C, CyclinD1, CDC25A and PCNA mRNA expression of cholangiocarcinoma, with kind of individual bile duct cancer tissue and normal tissue Survivin mRNA relative expression levels were 0.6238 ± 0.080,0.5693 ± 0.0443 and 0.5684 ± 0.0317, Survivin mRNA expression between the three groups showed a downward trend; cholangiocarcinoma, the same kind of individual cancer tissue and normal bile duct tissue VEGF-C mRNA relative expression levels were 0.6930 ± 0.1078,0.6270 ± 0.0664 and 0.6105 ± 0.0577, VEGF-C mRNA expression between the three groups showed a downward trend; cholangiocarcinoma, the same kind of self- bile duct cancer tissue and normal tissue CyclinD1 mRNA relative expression levels were 0.6288 ± 0.0942,0.5608 ± 0.0577 and 0.5845 ± 0.0310, CyclinD1 mRNA expression between the three groups showed a downward trend; cholangiocarcinoma, the same kind of individual cancer tissue and normal bile duct tissues CDC25A mRNA relative expression levels were 0.5937 ± 0.0716,0.5229 ± 0.0575 and 0.5135 ± 0.0780, CDC25A mRNA expression between the three groups showed a downward trend; cholangiocarcinoma, the same kind of individual cancer tissue and normal bile duct organization PCNA mRNA relative expression levels were 0.6303 ± 0.0773,0.5692 ± 0.0408 and 0.5605 ± 0.0331, PCNA mRNA expression between the three groups also showed a downward trend. Conclusion Survivin, VEGF-C, CyclinD1, CDC25A and PCNA mRNA and protein expression in cholangiocarcinoma consistent, highly expressed. These changes in gene and protein expression may be caused by the development of the molecular mechanisms of cholangiocarcinoma.
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