Neutrophil elastase induces cell proliferation and migration by the release of TGF-alpha, PDGF and VEGF in esophageal cell lines.

Rapid regrowth or recurrent growth of occult cancer cells are often observed after esophagectomy or postoperative complications. In order to clarify the mechanism of such oncological circumstances, we focused on neutrophil elastase (NE), which degrades a broad spectrum of extracellular matrix and cell surface proteins. In the present study, we demonstrated that NE stimulated the growth of all of the five esophageal cell lines (TE-1, -7, -8, -12 and -13) by MTT assay and promoted cell invasion by cell migration assay. Pro-transforming growth factor-alpha (pro-TGF-alpha) from the cell membrane was released to the culture medium as a mature form after treatment with 5 microg/ml NE, and it reached the maximum level of 153% compared to the control values at 15 min of treatment in TE-13 cells. The phosphorylation of epidermal growth factor receptor (EGFR) rapidly occurs after treatment with NE and triggered the extracellular signal-regulated kinases 1 and 2 (ERK) signaling pathway. Moreover, NE induced release of platelet-derived growth factor-AA (PDGF-AA), PDGF-BB and vascular endothelial growth factor (VEGF) to 141.9, 227.7, and 171.6% of the control values, respectively. A specific NE inhibitor, sivelestat, significantly inhibited the NE-induced cell proliferation, cell invasion and subsequently inhibited the signal transduction pathway. Furthermore, sivelestat significantly inhibited NE-induced release of TGF-alpha, PDGF-AA, PDGF-BB and VEGF in the medium in TE-13 esophageal carcinoma cells. These results strongly indicate that NE released from activated neutrophils stimulates the growth and progression of esophageal cancer cells by releasing the growth factors on the cell surface and that sivelestat, a specific NE inhibitor, blocks these processes. Furthermore, we postulate that postoperative administration of sivelestat might be useful as a new molecular-targeting cancer therapy as well as for the treatment of postoperative respiratory complications.

Expression of gastric mucin MUC5AC and gastric transcription factor SOX2 in ampulla of vater adenocarcinoma: comparison between expression patterns and histologic subtypes.

The purpose of this study was to examine the expression pattern of MUC5AC and SOX2 in ampulla of vater adenocarcinoma and evaluate the association between expression of these gastric epithelial markers and the histologic phenotype of ampulla of vater carcinoma. Six surgically resected samples of ampulla of vater adenocarcinoma, including four intestinal type carcinomas and two pancreatobiliary type carcinomas, were studied. We performed immunohistochemistry with a monoclonal antibody against MUC5AC and a polyclonal anti-SOX2 antibody. In two of the four intestinal type carcinomas, MUC5AC and SOX2 were focally expressed in the superficial neoplastic mucosa. However, in the centre of the tumour and in other invasive lesions, including vascular invasive lesions and metastatic lymph nodes, neither MUC5AC nor SOX2 was expressed. In contrast, in both pancreatobiliary type carcinomas, expression of MUC5AC and SOX2 was maintained or increased in invasive lesions. Our immunohistochemistry data suggest that MUC5AC and SOX2 are associated with the pancreatobiliary phenotype of ampulla of vater carcinoma and involved in later events in carcinogenesis, such as invasion and metastasis.

The expression profiles of orotate phosphoribosyltransferase and dihydropyrimidine dehydrogenase in gastric cancer and their clinical significance.

Orotate phosphoribosyltransferase (OPRT) is an enzyme that causes the activation of 5-fluorouracil (5-FU). Dihydropyrimidine dehydrogenase (DPD) is known to catabolize 5-FU, which is widely used in chemotherapeutic treatments for patients with a variety of malignant tumors including gastric and colorectal cancer. The expression and activities of these two enzymes therefore play important roles in the response of cancer patients to chemotherapy. However, little is known about the expression of these enzymes in gastric cancer. In the present study, we further elucidate the expression patterns of ORPT and DPD and their clinicopathological significance by immunohistochemical analysis in 221 and RT-PCR in 36 gastric cancer samples. The expression of OPRT by immunohistochemical analysis was detected in 117 (52.9%) cases, whereas DPD was detected in 66 (29.9%) cases. Moreover, the level of expression of OPRT was found to correlate with the depth of tumor invasion and a poorer prognosis. Although the mRNA and protein expression of OPRT and DPD levels did not correlate, an inverse correlation in the expression of OPRT and DPD was observed by RT-PCR. The survival benefit of post-operative adjuvant chemotherapy could not be confirmed in our present analysis. However, among the patients who had received such treatment with 5-FU or its derivatives, the prognosis in cases with low DPD levels was better than that in cases with high DPD expression by immunohistochemical analysis. These results indicate that the expression of OPRT and DPD are important predictors of both survival and the response to adjuvant chemotherapy in gastric cancer patients.

[Clinical evaluation, including QOL of TS-1 for patients with locally advanced or recurrent gastric cancer at ambulatory setting].

PURPOSE: Forty-nine patients with locally advanced or recurrent gastric carcinoma were treated with a novel 5-FU derivatives, TS-1, in an ambulatory setting. The response rate and adverse effect as well as patients' QOL were evaluated. RESULTS: The overall response rate was 38.8% (19/49). Partial response (PR) was obtained in 3 (27%) of 11 primary lesions of the stomach, in 10 (48%) of 21 lymph node metastases, in 6 (40%) of 15 liver metastasis, and in 4 (33%) of 12 peritoneal disseminations, respectively. The average response period was 222.2 days and the 50% survival period was 382 days. In addition, patients' QOL, evaluated by questionnaire, was maintained relatively well during treatment. Conversely, the adverse effects (greater than grade 3) were bone marrow suppression in 3 cases and toxic dermatitis in 1 case, respectively. CONCLUSION: Taken together it is reasonable to conclude that TS-1 is safe and effective for patients with locally advanced or recurrent gastric carcinoma in an ambulatory setting, and is promising as a first line treatment in the general hospital.

Dihydropyrimidine dehydrogenase and orotate phosphoribosyltransferase in esophageal cancer patients: Correlation with clinicopathological factors and prognosis.

Thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phosphoribosyltransferase (OPRT) are fluoropyrimidine metabolic enzymes which play important roles in the response of cancer patients to chemotherapy. In esophageal cancer, little is known about the relationship between the expression of these enzymes and corresponding clinicopathological features. In the present study, TS, DPD and OPRT expression levels were evaluated in 72 resected esophageal cancer specimens using immunohistochemistry. The relationship between enzyme expression and clinicopathological features was assessed using Fisher's exact test or the χ2 test (categorical variables), or the Mann-Whitney rank-sum test (continuous variables). Survival curves were calculated using the Kaplan-Meier method, and differences evaluated using the log-rank test. The Cox proportional hazards model was also used. High DPD expression was associated with depth of invasion, nodal status, tumor stage, lymphatic invasion and venous invasion (P<0.001, P=0.004, P<0.001, P=0.006, P=0.038, respectively), as well as with decreased patient survival (P=0.007). In patients receiving adjuvant chemotherapy, low DPD expression did not significantly improve recurrence-free survival. OPRT was particularly expressed in esophageal cancer cells as compared to normal squamous cells. High OPRT expression was associated with depth of invasion and venous invasion (P=0.006, P=0.003, respectively). To conclude, in esophageal cancer DPD expression was associated with tumor progression and prognosis, and OPRT expression was correlated with carcinogenesis and tumor progression.

Histopathologic evaluation of stepwise progression of pancreatic carcinoma with immunohistochemical analysis of gastric epithelial transcription factor SOX2: comparison of expression patterns between invasive components and cancerous or nonneoplastic intraductal components.

OBJECTIVES: The purpose of this study was to perform histopathologic and immunohistochemical analyses of gastric transcription factor SOX2 and gastric mucin MUC5AC to better understand the stepwise progression of pancreatic carcinoma. METHODS: Twenty-eight representative sections from 14 surgically resected pancreatic carcinomas were assessed microscopically. Sites of pancreatic intraepithelial neoplasia (PanIN) were counted, and histologic subtypes of invasive ductal carcinoma (IDC) were determined. The expression of SOX2 and MUC5AC in PanIN and IDC was examined immunohistochemically. RESULTS: One hundred thirty-eight PanINs were identified. In 4 of the 14 cases, gradual transition from PanIN-1A to PanIN-3 was observed in a single duct, suggesting stepwise progression. The expression of MUC5AC increased with the progression of lesions from PanIN-1A to PanIN-3. SOX2 was expressed in only 6 of 107 early PanINs (5.8%). Out of 31 PanIN-3s, 7 were positive (22.6%), and SOX2 protein was localized in the nuclei of cells of the basal epithelium or in the vicinity of luminal necrosis. In addition, SOX2 was frequently and strongly expressed in poorly differentiated (57.1%) and neurally invasive (63.6%) components. CONCLUSIONS: The results of our histopathologic examinations suggest that PanIN progresses stepwise to IDC. Immunohistochemistry results suggest that SOX2 is involved in later events of carcinogenesis.