1. Expression Of PTTG In Gallbladder Carcinoma 2.The Effects Of PTTG Antisense CDNA On Gallbladder Carcinoma Cells

Objective 1. To investigate the expression of PTTG in human gallbladder carcinoma tissues and the correlation with angiogenesis and other clinicobiological behaviors; and to explore their correlation with bFGF and c-myc. 2. To explore the effects of PTTG antisense cDNA on proliferation and apoptosis in gallbladder carcinoma cells and sensitivity of gallbladder carcinoma cells to 5-FU.Methods The expressions of PTTG, bFGF and c-myc in 41 cases of human gallbladder carcinoma and 22 cases of chronic cholecystitis were detected with immunohistochemistry of SP. The microvessels were highlighted by immunohistochemical staining (SP method) to detect antigen of CD34. Angiogenesis was represented by microvessel density (MVD). The recombinant vector containing PTTG antisense cDNA was transfected into GBC-SD gallbladder cancer cells. The positive cell clones were screened by G418. PTTG protein expression was detected by Western blot analysis. The changes of cell proliferation and cell relative viability after treatment with different concentrations of5-FU were assessed by MTT method, the dose-effects curves were obtained and IC50S were calculated. Flow cytometry was used to demonstrate discrepancy of apoptotic rates. All the statistical analyses were performed by using SPSS 10 software. Statistical comparisons were made by using Student's Mest, x²-test, F-test, or Kruskal-Wallis H test respectively. Spearman's non-parametrical correlation analysis was used. P<0.05 was considered significant. IC₅₀S were calculated by using SPSS 10 software.Results Expressions of PTTG, bFGF, c-myc were 82.9%, 75.6%, 63.4% respectively, which were significantly higher than those in the chronic cholecystitis (P=0.002, 0.006, 0.002). The expression of PTTG was significantly associated with clinical stages and lymph node metastasis status (P-0.025, 0.007), but not with histological differentiation (P=0.144). The expression of bFGF was significantly correlated with clinical stages and histological differentiation ( P=0.019,0.015 ) , but not with lymph node metastasis status (P=0.081). There was a significant correlation between the expression of PTTG and bFGF (r=0. 648, P=0.000) . The expression of c-myc was significantly correlated with lymph node metastasis status (P=0.032) ,but not with histological grades and clinical stages (P=0.127, 0.198). There was a significant relation between the expression of PTTG and c-myc 0=0. 463, p=0.002) . None of them had relation with age, sex, histological type and cholelithiasis. The value of MVD in the gallbladder carcinoma was significantly higher than that in the chronic cholecystitis(t=3.684,P=0.001). The expression of PTTG, bFGF and c-myc were respectively correlated with MVD in gallbladder carcinoma (P=0.000,0.000,0.02). MVD in the gallbladder carcinoma was significantly associated with clinical stages and lymph node metastasis status (p=0.024, 0.007), but not with age, sex, histological type,...