Prognostic significance of epithelial-mesenchymal transition-related markers in extrahepatic cholangiocarcinoma: comprehensive immunohistochemical study using a tissue microarray.

BACKGROUND: Epithelial-mesenchymal transition (EMT) is characterised by the loss of cell-to-cell adhesion and gaining of mesenchymal phenotypes. Epithelial-mesenchymal transition is proposed to occur in various developmental processes and cancer progression. 'Cadherin switch', a process in which cells shift to express different isoforms of the cadherin transmembrane protein and usually refers to a switch from the expression of E-cadherin to N-cadherin, is one aspect of EMT and can have a profound effect on tumour invasion/metastasis. The aim of this study was to investigate the clinicopathological significance of EMT-related proteins and cadherin switch in extrahepatic cholangiocarcinoma (EHCC). METHODS: We investigated the association between altered expression of 12 EMT-related proteins and clinical outcomes in patients with EHCC (n=117) using immunohistochemistry on tissue microarrays. RESULTS: Univariate and multivariate analyses revealed that, in addition to N classification (P=0.0420), the expression of E-cadherin (P=0.0208), N-cadherin (P=0.0038) and S100A4 (P=0.0157) was each an independent and a significant prognostic factor. We also demonstrated that cadherin switch was independently associated with poor prognosis (P=0.0143) in patients with EHCC. CONCLUSIONS: These results may provide novel information for selection of patients with EHCC who require adjuvant therapy and strict surveillance.

Association of CD8+ T cell infiltration in oesophageal carcinoma lesions with human leucocyte antigen (HLA) class I antigen expression and survival.

Oesophageal cancer is one of the most aggressive tumours with a poor prognosis. However, little is known about the immune response in the tumour microenvironment. To investigate the role of immunosurveillance in the clinical course of oesophageal squamous cell carcinoma, 98 formalin-fixed, paraffin-embedded primary tumours were analysed using immunohistochemical methods for human leucocyte antigen (HLA) class I heavy chain and ß2-microglobulin expression and for CD4-, CD8- and CD57-positive cell infiltration. HLA class I expression of tumour cells was correlated positively with infiltration of CD8(+) T cells into the cancer nest, but not with the clinical course of disease. However, CD8(+) and CD4(+) T cell infiltration was correlated with prognosis. These results suggest that tumour antigen-specific cellular immune response plays a role in the clinical course of the disease and that HLA class I antigen expressed on tumour cells contribute to this association most probably by mediating the interactions between tumour cells and CD8(+) T cells.

Infiltrating regulatory T cell numbers is not a factor to predict patient's survival in oesophageal squamous cell carcinoma.

CD4/8 status has been previously reported to be a critical factor in the prognosis of oesophageal squamous cell carcinoma (OSCC). In the current study, we investigated the effect of regulatory T cells (Treg; Foxp3+ lymphocytes) on the status of CD4+ and CD8+ T cells in 122 patients with OSCC. Immunohistochemical analysis of Treg was performed using an antibody against Foxp3. The survival rate for low Foxp3 patients was significantly lower than for high Foxp3 patients (P=0.0028 by log-rank test), but Foxp3 status did not significantly correlate with prognosis in CD4/8(+/+) patients or CD4/8(+/-) or (-/+) patients (P=0.5185 and 0.8479, respectively, by log-rank test). We also found that Foxp3 status correlated with CD4/8 status (P=0.0002 by chi2 test) and that the variance of CD8/CD4 ratio in patients with low Foxp3 was larger than in patients with high Foxp3 (P<0.0001 by F-test). Thus, the results do not support the idea that Treg suppress anti-tumour immunity in patients with OSCC. Rather, the CD8/CD4 ratio and CD4/8 status appear to be critical factors in anti-tumour immunity. Furthermore, Treg numbers correlate with both the CD8/CD4 ratio and the CD4/8 status, suggesting that Treg number is not a factor to predict patient's survival in OSCC.

Changes of biochemical markers during fracture healing.

The aim of this study was to evaluate the changes of biochemical markers during fracture healing in patients with osteoporosis. The study included 26 patients; 9 underwent hip hemiarthroplasty (mean age +/- SD: 71.0 +/- 10.2 years, group EN) for femoral neck fractures. 7 underwent osteosynthesis (75.3 +/- 8.2 years, group OS) for trochanteric fractures, and 10 subjects had spinal compression fractures (68.2 +/- 12.0 years, group CO). No operative procedures were performed in group CO. Urinary pyridinoline (Pyr), deoxypyridinoline (Dpyr) by high performance liquid chromatography (HPLC), Crosslaps by both enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) (CTx-ELISA and CTx-RIA) and serum N-terminal mid-fragment osteocalcin (OCN-Mid) by ELISA were analyzed at the time of admission and at weeks 1, 2, 4, 8 and 24 after operation or, in the case of group CO, after admission. As a whole, bone resorption markers started to increase from week 1, with various peak values between weeks 4 and 8 depending upon the particular marker, but returned to the initial vales at week 24. OCN-Mid started to increase from week 8 and remained at elevated levels at week 24. In groups EN and OS, bone resorption markers changed in the same manner as they did as a whole group. OCN-Mid did not change in group EN, although it increased significantly from week 8 in group OS. No biochemical markers changed significantly in group CO. In conclusion, bone resorption was accelerated at an early stage due to acute osteonecrosis or bed rest, followed by bone formation due to callus or mechanical stress later on. As far as bone resorption markers are concerned, 24 weeks are enough to eliminate the effect of fracture.