Prognostic significance of preoperative inflammatory response biomarkers in patients undergoing curative thoracoscopic esophagectomy for esophageal squamous cell carcinoma.

BACKGROUND: Recent studies have revealed significant relationships between the lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) and survival in various cancers. The purpose of this study was to confirm whether the LMR, NLR, and PLR have prognostic values, independent of clinicopathological criteria, in patients undergoing curative resection for esophageal cancer. METHODS: The LMR, NLR and PLR were calculated in 147 consecutive patients who underwent curative esophagectomy between January 2006 and December 2014. Receiver operating characteristics (ROC) curve analysis was conducted to identify the optimal cutoff values of each biomarkers. RESULTS: In multivariate analysis for cancer-specific survival (CSS), pTNM stage (p < 0.0001) and low LMR (p = 0.0081) were selected as independent prognostic factor. Similarly, pTNM stage(p < 0.0001) and low LMR (p = 0.0225) were found to be independent prognostic factor for overall survival (OS). There was no significant relationship between LMR, NLR and PLR and survival in patients with stage I or II, however, significant relationships between LMR and CSS or OS were observed in patients with stage III esophageal cancer. CONCLUSIONS: LMR can be used as a novel predictor of postoperative CSS and OS in patients with esophageal cancer and that it may be useful in identifying patients with a poor prognosis even after radical esophagectomy.

Impact of inflammation-based prognostic score on survival after curative thoracoscopic esophagectomy for esophageal cancer.

BACKGROUND: Despite recent improvements in early detection, progress in surgical techniques, and development of chemoradiation therapies, prognosis of esophageal cancer remains poor. The aim of the present study was to assess whether Glasgow Prognostic Score (GPS), an inflammation-based prognostic score, has prognostic value independent of conventional clinicopathological criteria in patients undergoing curative resection for esophageal cancer, even in elderly patients. METHODS: We retrospectively reviewed the database of 141 consecutive patients with histologically verified esophageal squamous cell carcinoma who underwent potentially curative surgery in our institute, between January 2006 and December 2014. GPS and neutrophil lymphocyte ratio (NLR) were calculated. RESULTS: On multivariate analysis, TNM stage (p < 0.0001) and GPS (p = 0.041) were independently associated with worse prognosis in overall patients with esophageal cancer. Multivariate analysis evaluated the prognostic factors in two different patient groups: patients younger than 70 years (non-elderly) and those aged 70 years or more (elderly). Multivariate analysis demonstrated that TNM stage (p = 0.0003) was an only independent risk factor for a worse prognosis among non-elderly group. Meanwhile, multivariate analysis demonstrated that TNM stage (p = 0.001) and GPS (p = 0.043) were the independent risk factor for a worse prognosis among elderly group. CONCLUSION: The present study demonstrated that GPS is associated with prognosis and can be considered as an independent prognostic marker in patients who underwent esophagectomy. Moreover, the GPS has the advantage of being simple to measure, routinely available and well standardized. But the present study failed to confirm the NLR as a significant predictor of survival following resection for esophageal cancer.

Inoculation of human interleukin-17 gene-transfected Meth-A fibrosarcoma cells induces T cell-dependent tumor-specific immunity in mice.

OBJECTIVE: The biological activities of interleukin-17 (IL-17), a newly cloned cytokine, have not been fully elucidated. The present study was designed to assess the in vitro and in vivo effect of transfecting the IL-17 gene into tumor cells. METHODS: A complementary DNA (cDNA) encoding human IL-17 (hIL-17) was obtained by polymerase chain reaction amplification from the human CD4+ T cell cDNA library and inserted into the plasmid pRc/cytomegalovirus to construct an expression vector for the hIL-17 gene. Murine Meth-A fibrosarcoma cells were transfected with the hIL-17 gene using the lipofectin method. The hIL-17 gene-expressing clone (Meth-A/IL-17) was selected and analyzed for cytokine expression by Northern blot. RESULTS: There was no significant difference in the in vitro proliferation rate among parent Meth-A, cells transfected with vector alone and Meth-A/IL-17 cells. When the tumor cells were transplanted subcutaneously into BALB/c nude (nu+/nu+) mice, there was no difference in in vivo growth rates among the three cell lines. Challenge with tumor cells in conventional BALB/c mice, however, resulted in the rejection of Meth-A/IL-17 cells, but the other two lines did grow. After immunization with Meth-A/IL-17 cells, the mice were rechallenged by parent Meth-A or syngeneic MOPC-104E plasmacytoma cells; the immunized mice rejected the Meth-A cells, but not the MOPC-104E cells. Injecting the anti-thy 1,2 (CD90), anti-CD4 or anti-CD8 monoclonal antibody into conventional BALB/c mice resulted in the resumption of in vivo growth of Meth-A/IL-17 cells, but injecting the anti-asialo GM1 antibody did not. Furthermore, flow cytometric analysis demonstrated a significant increase in the expression of major histocompatibility complex (MHC) class I and class II antigens and lymphocyte function-associated antigen-1 on Meth-A/IL-17 cells. CONCLUSION: Meth-A cells transfected with the hIL-17 gene can induce tumor-specific antitumor immunity by augmenting the expression of MHC class I and II antigens, and both CD4+ and CD8+ T cells may play important roles in inducing antitumor immunity, suggesting the possibility of developing a tumor vaccine incorporating IL-17-transfected tumor cells.

Heterogeneic distribution of thymidine phosphorylase between primary tumors and metastatic lesions of human pancreatic ductal carcinoma: implications for the efficacy of chemotherapy with 5-FU or its derivatives.

PURPOSE: It has been suggested that the expression of thymidine phosphorylase (TdRPase) correlates with the malignant potential of various cancers, but its involvement in human invasive ductal carcinoma (IDC) of the pancreas has not been reported. In the present study, the distribution and clinical significance of TdRPase in IDCs and benign diseases of the pancreas were assessed, especially in relation to the efficacy of chemotherapy with 5-FU or its derivatives. METHOD: The expression of TdRPase in 148 specimens of pancreatic IDCs (66 primary lesions, 46 nodal lesions and 36 distant metastases from 126 patients) and in 24 specimens of benign diseases (4 cystadenomas, 3 hyperplasias, and 17 chronic pancreatitises) was examined by immunohistochemical staining with anti-TdRPase monoclonal antibody and evaluated in terms of three grades of immunoreactivity: negative 0, low 1, or high 2. RESULTS: Positive TdRPase staining (low and high immunoreactivity) was detected in 71% (47/66) of the primary lesions, in 46% (21/46) of the involved nodes, in 53% (19/36) of various lesions of distant metastasis, and in 37% (9/24) of the benign diseases. The staining intensity was significantly higher in the IDC tissues than in the benign disease tissues, and significantly lower in the metastatic lesions than in the primary lesions. TdRPase reactivity did not correlate with the survival rate in both resectable and unresectable IDCs. In patients with both primary tumor and nodal involvement, however, high TdRPase activity in involved nodes was significantly associated with a poor prognosis. On the other hand, although adjuvant chemotherapy was found to improve the survival of patients, TdRPase activity in the tumor did not show any significant relationship with the efficacy of chemotherapy with 5-FU or its derivatives. CONCLUSIONS: The present study suggested that in pancreatic IDC the activity of TdRPase in primary lesions is different from that in metastatic lesions, and that DNA is synthesized mainly through the salvage pathway in primary lesions and through a de novo pathway in metastatic lesions. This may be one of the reasons for the heterogeneity in chemosensitivity of human pancreatic IDC.

Plant polysaccharide PSK: cytostatic effects on growth and invasion; modulating effect on the expression of HLA and adhesion molecules on human gastric and colonic tumor cell surface.

PSK is a plant polysaccharide widely used for cancer immunotherapy in Japan and other Asian countries. It is considered that its antitumor effect is derived from its immunomodulating activity on the tumor-bearing host. The present study was designed to assess the direct action of PSK on in vitro proliferation and invasion of human KATO-3 gastric and Colo205 colonic cancer cell lines, and the expression of surface molecules such as HLA and adhesion molecules on these cells. The in vitro growth of KATO-3 cells was significantly inhibited by 100 micrograms/ml of PSK 48 hrs after culture initiation, and that of Colo205 was significantly inhibited by 10 and 100 micrograms/ml of PSK 24 hrs after culture initiation. The effect of PSK on the in vitro invasion of the tumor cells, assessed with a Matrigel invasion chamber, revealed that invasion of KATO-3 and Colo205 cells was inhibited by more than 10 micrograms/ml and more than 5 micrograms/ml of PSK, respectively. KATO-3 cells expressed HLA-ABC, HLA-A2/A28, HLA-DR very weakly, at almost baseline levels, but HLA-B27, B2-microglobulin and HLA-DQ were expressed at various levels. After treatment of KATO-3 cells with PSK, the expression of HLA-B27 and beta 2-microglobulin was significantly enhanced. Colo205 cells expressed all class-I antigens tested in this study at different levels, but class-II antigens at almost baseline levels. PSK also enhanced the expression of class-I antigens on Colo205 cells. ICAM-1 was expressed on KATO-3, but not on Colo205. The expression of ICAM-1 was enhanced to a greater extent by treatment with 10 micrograms/ml than with 100 micrograms/ml of PSK. Adenocarcinoma antigen AC-81 was strongly expressed on both cell lines, but PSK-treatment significantly enhanced its expression. These results suggested that enhancement of HLA class-I expression on tumor cells after PSK treatment may be one of the mechanisms responsible for the induction of anti-tumor immunity by PSK.

Reduced invasiveness and metastasis of Chinese hamster ovary cells transfected with human interleukin-17 gene.

Human IL-17 (hIL-17) stimulates epithelial, endothelial, fibroblastic cells and macrophages to secrete various cytokines. The present study was designed to assess the effects of the transfection of the hIL-17 gene in Chinese hamster ovary (CHO) cells. A complementary DNA (cDNA)-encoding hIL-17 was obtained by polymerase chain reaction (PCR) amplification from human CD4+ T-cell cDNA and inserted into the plasmid pRc/CMV to construct an expression vector for hIL-17. CHO cells were transduced with hIL-17 DNA-carrying cytomegalovirus (CMV)-based retroviral vectors. A clone with a high mRNA expression of hIL-17 (CHO/IL-17) was selected by Northem blotting. There was no significant difference in the in vitro growth of cells among parent CHO cells, vector-only transfected cells (CHO/neo) and CHO/IL-17 cells. A Matrigel invasion chamber assay, however, demonstrated significantly lower invasiveness by CHO/IL-17 cells than by either the parent CHO or the CHO/neo cells. There was no difference in the in vivo growth among the cells, when subcutaneously transplanted into nude mice. When injected into the tail vein, however, the number of metastatic nodules in the lungs of CHO/IL-17-injected mice was significantly smaller than that of CHO- or CHO/neo-injected mice. Furthermore, NK activity of spleen cells was significantly higher in nude mice transplanted with CHO/IL-17 cells than in mice transplanted with parent CHO or CHO/neo cells. In conclusion, the hIL-17-gene-transfected CHO cells showed a significantly lower metastatic potential to the lung by directly modulating the invasiveness and metastasis of CHO cells as well as by enhancing NK activity.

Induction of tumor-specific antitumor immunity after chemotherapy with cisplatin in mice bearing MOPC-104E plasmacytoma by modulation of MHC expression on tumor surface.

Chemotherapy sometimes results in induction of specific antitumor immunity. We investigated the mechanisms responsible for the induction of antitumor immunity in mice bearing MOPC-104E plasmacytoma after chemotherapy with cisplatin (CDDP), especially the effects of CDDP on the expression of MHC on the tumor surface. BALB/c mice were subcutaneously (s.c.) inoculated with MOPC-104E cells on day 0, then intravenously (i.v.) treated with CDDP at 3.6 mg/kg on day 7. The tumors disappeared completely on day 35 and the mice rejected a second challenge with MOPC-104E, but did not reject syngeneic Meth-A fibrosarcoma. The tumors did not regress, however, when MOPC-104E was s.c. transplanted in nude mice, or when anti-T-cell monoclonal antibodies were i.v. injected into BALB/c mice before CDDP treatment on day 6. To determine which of the mice or tumor were affected by CDDP, BALB/c mice were inoculated with CDDP-treated (12.5 micrograms/ml for 3 hours in vitro) MOPC-104E cells on day 0, 7 and 14. The potential to reject MOPC-104E was lower in mice immunized with ethanol-treated MOPC-104E cells than in those immunized with CDDP-treated cells. CDDP-treated or -untreated MOPC-104E cells ultrasonicated and fractionated into soluble and insoluble fractions by centrifuging, also induced antitumor immunity. Flow cytometry demonstrated that the expression of MHC-class-I antigens H-2Dd and H-2Kd was enhanced after CDDP-treatment, but that of class-II antigens I-Ad and I-Ed was not, suggesting that CDDP induced tumor-specific antitumor immunity by enhancing the expression of MHC-class-I antigens.

Intraperitoneally implanted artificial pancrease with transkaryotic beta-cells on microcarrier beads in a diffusion chamber improves hyperglycemia after 90% pancreatectomy in rats.

BACKGROUND: Considering the difficulties in pancreas transplantation, the development of an artificial pancreas would be of great value. We have established a transkaryotic artificial beta-cell line, CHO/I, produced by transfecting the human proinsulin gene into the Chinese hamster ovary (CHO) cell line. The present study was designed to assess the value of an artificial pancreas using a diffusion chamber containing CHO/I cells. MATERIALS AND METHODS: Wistar rats rendered diabetic by 90% pancreatectomy were treated by implanting a diffusion chamber containing CHO/I cells cultured on microcarrier beads. RESULTS: The diffusion chamber containing microcarrier beads produced 100-folds more CHO/I cells than the chamber alone, as calculated from the secreted IRI in vitro. When diffusion chambers containing CHO/I cells on microcarrier beads were implanted into the peritoneal cavity of the 90% pancreatectomized rats, the fasting serum IRI level increased and the fasting blood glucose decreased to the normal level for 12 weeks. An intraperitoneal glucose tolerance test demonstrated, however, that the diffusion-chamber-implanted rats did not respond to glucose loading. CONCLUSION: An artificial pancreas using a diffusion chamber containing human proinsulin gene transfected cells might be a promising model for future clinical application.

Comparative significance of p53 and WAF/1-p21 expression on the efficacy of adjuvant chemotherapy for resectable invasive ductal carcinoma of the pancreas.

p53 tumor-suppressor gene has a dual role as a trigger of apoptosis and as an initiator of DNA repair. The cyclin-dependent kinase inhibitor WAF/1-p21 is induced by wild-type p53 and has been implicated as a downstream mediator of the growth-suppressing and apoptosis-promoting function of wild-type p53, suggesting an impact on the effectiveness of chemotherapy. This study was designed to assess the significance of p53 and WAF/1-p21 expression in the prognosis of patients and the efficacy of adjuvant chemotherapy for resectable invasive ductal carcinoma (IDC) of the pancreas. A total of 58 patients with primary IDC of the pancreas underwent pancreatectomy between 1982 and 1996: 28 patients underwent surgery alone, and 30 patients received postsurgical adjuvant chemotherapy. p53 and WAF/1-p21 were stained immunohistochemically with anti-p53 monoclonal antibody (mAb) and anti-WAF/1-p21 mAb. p53 was positively expressed in 29 (50%) of 58 primary lesions, and p21 was expressed in 24 (41%) lesions; however, p21 expression did not necessarily correlate with p53 expression. The survival curve of the patients with p53(+) IDC was significantly lower than that of those with p53(-) IDC, and p21(+) patients showed a higher survival curve than did p21(-) patients, but this difference was not statistically significant. When p53 and p21 expression were analyzed in combination, the patients with p53(+)p21(-) IDC were found to have a significantly poorer prognosis than others. On the other hand, the survival curve of the adjuvant chemotherapy group was also higher than that of the surgery-alone group, but this difference was not significant. In a multivariate analysis, p21 expression was a significantly low risk factor for death due to IDC overall, and adjuvant chemotherapy was found to decrease the risk of death from IDC in p53(+) patients. Evaluation of expression of p53 and WAF/1-p21 may be beneficial in the prediction of the patient's prognosis as well as prediction of the effects of adjuvant chemotherapy in pancreatic cancer patients.

Second malignancies after a gastrectomy for gastric cancers: the effects of adjuvant therapies.

Second malignancy after gastrectomy has become one of the important issues in the management of gastric cancer patients. However, the effect of adjuvant therapies has never been surveyed, although there have been many reports on second malignancy after chemotherapy of other malignancies. Between 1979 and 1995, 574 patients with gastric cancer underwent gastrectomies in our department. 563 patients (98.1%) were completely followed-up, and the incidence of second malignancy after gastrectomy was assessed with special emphasis on the effect of adjuvant therapies, namely benefits versus carcinogenesis. The overall postsurgical survival rate was 62.5%, and the survival rate of patients with stage 2-4 cancer was significantly higher in the adjuvant therapy groups than in the surgery alone group. The overall incidence rate of a second malignancy was 3.20% (18/563), including 4 colorectal, 3 liver, 3 head and neck, and 8 other malignancies, and 3.90% (9/231) for the surgery alone group, 1.94% (3/155) for the chemotherapy group, 3.68% (6/163) for the chemoimmunotherapy group, and 0% (0/14) for the immunotherapy alone group. There were no statistical differences in the incidence rates between these 4 groups. The mean duration until the occurrence of second malignancy was 5.1 years for the surgery alone group, 8.6 years for the adjuvant therapy group (9.3 years for chemotherapy and 8.1 years for chemoimmunotherapy; p < 0.1, surgery alone vs. adjuvant therapy). The overall mean actuarial risk of developing a second malignancy was 6% at 5 years and 18% at 10 years. Multivariate analysis demonstrated that chemotherapy decreased the risk of death due to gastric cancer (risk ratio = 0.67, p = 0.017) and the risk of a second malignancy (risk ratio = 0.52, p = 0.261). Oral fluoropyrimidines were found to be particularly effective at decreasing the risk of death due to gastric cancer (risk ratio = 0.66, p = 0.012) and a second malignancy (risk ratio = 0.38, p = 0.136). However, intravenous chemotherapy was demonstrated to have no influence on survival (risk ratio = 0.93, p = 0.692) or a second malignancy (risk ratio = 0.58, p = 0.500). There was no evidence of an increased risk for a second malignancy following adjuvant chemotherapy. Adjuvant chemotherapy with oral fluoropyrimidines was suggested to contribute to the improved survival of patients with stage 2-4 gastric cancer, and to have a decreased the risk of a second malignancy.

Human proinsulin gene-transfected Chinese hamster ovary cells secrete immunoreactive insulin.

BACKGROUND: Considering the difficulties in pancreas transplantation, the development of an artificial pancreas can be one of the new approaches. The present study was designed to assess whether or not Chinese hamster ovary (CHO) cells, which were transfected with the human proinsulin (hPI) gene, secrete immunoreactive insulin (IRI) and respond to glucose loading. MATERIALS AND METHODS: A complementary DNA encoding hPI was obtained by polymerase chain reaction amplification from human pancreatic tissue and was inserted into the plasmid pcDNA I/NEO to construct an expression vector for the hPI gene. CHO cells were transfected with hPI gene using lipofectin, and the hPI gene-expressing clones (CHO/I) were selected. RESULTS: Five clones of CHO/I cells, releasing IRI into the culture supernatant, were separated. Immunohistochemistry with a monoclonal antibody demonstrated the IRI in the cytoplasm of CHO/I cells, and transmission electron microscopic examination demonstrated the prominently developed mitochondria, but no secretion granules. ELISA assay demonstrated the secretion of IRI into the culture supernatant of CHO/I, but CHO/I cells did not respond to the glucose loading. When CHO/I cells were transplanted subcutaneously into the back of nude mice, the growing tumors secreted IRI. CONCLUSIONS: These results demonstrate that the hPI gene can be transfected into mammalian cells and function in vivo, and suggest that this kind of gene technology may be applicable in the development of an artificial pancreas.

p53 expression affects the efficacy of adjuvant chemotherapy after resection of invasive ductal carcinoma of the pancreas.

p53 tumor suppressor gene has a dual role as a trigger of apoptosis and as an initiator of DNA repair, suggesting its involvement in the mechanisms of drug resistance or chemosensitivity. The present study assessed the implication of p53 expression in the prognosis of patients and the efficacy of adjuvant chemotherapy for resectable invasive ductal carcinoma (IDC) of the pancreas. A total of 58 patients with primary IDC of the pancreas underwent pancreatectomy between 1982 and 1996: 28 patients received surgery alone and 30 patients received postsurgical adjuvant chemotherapy. p53 protein was stained immunohistochemically with anti-p53 monoclonal antibody. p53 was positively expressed in 29 out of 58 primary lesions (50%), and the survival curve of the patients with p53 (+) pancreatic cancer is lower than that of those with p53 (-) cancer. On the other hand, the survival curve of adjuvant chemotherapy group was also higher than that of surgery alone group, and furthermore, in patients with p53 (+) cancer, the survival curve of adjuvant chemotherapy group was significantly better than that of the surgery alone group. A multivariate analysis showed that p53 expression or adjuvant chemotherapy is not a significant risk-factor for prognosis, but that adjuvant chemotherapy is a significant risk factor for the patients with p53 (+) pancreatic cancer, which suggests that p53 expression affects the efficacy of chemotherapy. p53 expression may be beneficial as an indicator for introduction of adjuvant chemotherapy in pancreatic cancer patients.

Mechanisms of cytotoxic effects of heavy water (deuterium oxide: D2O) on cancer cells.

Heavy water (deuterium oxide: D2O) contains a neutron and a proton in its hydrogen atoms and shows a variety of biologic activities different from normal light water. In the present study the cytotoxic and cytostatic activity of D2O was assessed using a BALB/c-3T3 fibroblast cell line and four human digestive organ cancer cell lines, i.e. HepG2 hepatic, Panc-1 pancreatic, KATO-3 gastric and Colo205 colonic cancer cell lines. Against four cancer cell lines, D2O showed significant cytotoxic and cytostatic effects in a MTT assay and a Trypan blue dye exclusion assay, at concentrations higher than 30% D2O. These effects were time and dose dependent, and the IC50 after 72 h of culture ranged from 20 to 30% D2O in the Trypan blue dye exclusion assay and from 30 to 50% D2O in the MTT assay. By contrast, IC50 for the 3T3 fibroblast cell line after 72 h of culture was about 15% in the Trypan blue dye exclusion assay and 50% inhibition was not achieved in the MTT assay. Furthermore, D2O was found to significantly inhibit the invasion of tumor cells in a Matrigel invasion chamber assay at concentrations higher than 10% D2O. Incubation with D2O resulted in enlargement of cells, nuclear pyknosis and vacuolization, and immunostaining studies demonstrated that D2O treatment resulted in an increase in nuclear nick-end-labeling, which indicates DNA fragmentation, in KATO-3 and HepG2 cell lines. Furthermore, the nucleic acids and protein synthesis inhibition assay suggested that the inhibition of DNA synthesis may be one of the mechanisms responsible for the antitumor effects of D2O. Furthermore, oral administration of D2O resulted in a significant inhibition of the growth of Panc-1 tumor xenografted s.c. in nude mice, but survival was not prolonged. In conclusion, D2O has cytotoxic and cytostatic activities against human digestive organ cancer cell lines, and D2O may be a potential anticancer agent.

A Case of Multiple Juvenile Papillomatosis of the Breast and Its Immunohistochemical Pathology.

Juvenile papillomatosis(JP)is a benign proliferative disease of the breast affecting young women. Some patients with JP develop breast carcinoma. We recentlytreated and followed a case of JP in a 16-year-old female, and studied its immunohistochemical characteristics. The patient noticed a small tumor, 5 mm in diameter, in the periareolar region on the medial side of the right breast. She underwent an excisional biopsy and pathology demonstrated JP. However, 6 months later, two new lesions were discovered by routine echogram examination, and were later excised. Immunohistochemistry demonstrated that the JP tumors were negative for estrogen receptor, but positive for progesterone receptor, epidermal growth factor receptor and erbB-2. In addition, tests for p53 were negative and Rb protein was normally expressed. Immunohistochemical analysis suggested that an abnormality in estrogen receptor is important in the pathogenesis of JP, and that the patient had a relatively high risk of developing breast carcinoma, since her JP is multicentric and recurrent. The present study also suggests that immunohistochemical pathology may be beneficial in assessing the malignant potential of JP.

A quinolinone derivative, vesnarinone (OPC-8212), significantly inhibits the in vitro and in vivo growth of human pancreatic cancer cell lines.

A quinolinone derivative, vesnarinone, has been used as a cardiotonic agent. Previous studies have demonstrated that vesnarinone has potent antitumor activity. The present study was designed to assess the antitumor effects of vesnarinone on human pancreatic cancer cell lines in vitro and in vivo. The in vitro effects of vesnarinone on the human pancreatic cancer cell lines (PANC-1, MIA PaCa-2 and BxPC-3) were assessed by the MTT assay, the Trypan blue dye exclusion test and the Matrigel invasion chamber assay. The inhibition of in vivo tumor growth was evaluated on two human pancreatic cancer xenografts (BxPC-3 and SPa-1) transplanted s.c. into nude mice. The dose of vesnarinone for 50% inhibition of cell growth in a 7 day culture ranged between 10 and 20 micrograms/ml as verified by the Trypan blue dye exclusion test. The dose for 50% cytotoxicity after a 3 day culture in the MTT assay was 32 micrograms/ml for PANC-1 and 30 micrograms/ml for BxPC-3, but 50% cytotoxicity for MIA PaCa-2 was not achieved by the maximal dose of vesnarinone (50 micrograms/ml). Nomalsky optic microscopy and acridine orange staining demonstrated the vacuolization and crater-like changes in the cell nucleus after vesnarinone treatment. Moreover, staining of an apoptosis marker (Le(y) protein) and nick end-labeling increased. Vesnarinone also inhibited cancer invasion in the Matrigel invasion chamber assay. In vivo, BxPC-3 and SPa-1 were s.c. transplanted into the nude mice, and vesnarinone (5 or 50 mg/kg) was daily administered orally for 21 days. In both lines, vesnarinone at 50 mg/kg achieved significant inhibition. The present study suggests that vesnarinone may be a new therapeutic agent for pancreatic cancer.

p53 protein expression as prognostic factor in human pancreatic cancer.

Mutations in the p53 tumor suppressor gene are considered to play an important role in the carcinogenesis of pancreatic cancer. The present study was designed to assess the clinicopathological significance of the expression of p53 protein in human pancreatic cancer. A total of 64 specimens of pancreatic cancer (44 primary and 20 metastatic lesions) were obtained by surgery in our department between 1982 and 1995, and p53 protein was stained using an immunohistochemical staining method (strepto-avidin-biotin complex method) with 2 kinds of anti-p53 monoclonal antibodies (DO-1 and PAb240). DO-1-p53 was usually stained in the nucleus of cancer cells and was positively expressed in 35 out of 64 specimens (54.7%): 21 out of 44 primary lesions (47.7%), and 14 out of 20 metastatic lesions (70.0%). On the other hand, PAb240-p53 was stained in both the nucleus and cytoplasm in 45 out of 64 specimens (70.3%): 33 out of 44 primary lesions (75.0%) and 12 out of 20 metastatic lesions (60.0%). The survival rate of the patients with DO-1-p53 (+) pancreatic cancer after pancreatectomy was significantly lower than that of patients with DO-1-p53 (-) pancreatic cancer. On the other hand, there were no significant implications of PAb240-p53 protein expression on survival after pancreatectomy. With regard to the clinicopathological characteristics, the rate of DO-1-p53 protein expression was significantly higher in elderly patients (> or = 65 y.o.). As a result, DO-1-p53 protein expression may be a beneficial prognostic factor for patients with pancreatic cancer, and it is a factor independent of the stage and other clinicopathological factors.

Ventricular unloading and improvement in left ventricular function after angiotensin converting enzyme inhibition with enalapril in patients with chronic congestive heart failure.

To clarify how angiotensin converting enzyme inhibition affects left ventricular function through ventricular unloading, ventricular wall stress, ventricular volumes, and other cardiac indices and exercise tolerance were evaluated in 17 patients with mild to moderate chronic congestive heart failure before and after 3 months of treatment with enalapril. Echocardiographic examination revealed that treatment with this angiotensin converting enzyme inhibitor resulted in significant reductions in end-systolic wall stress (117 +/- 25 to 89 +/- 28 g/cm2, p < 0.01) and left ventricular volume indices (end-diastolic: 163 +/- 56 to 143 +/- 60; end-systolic 99 +/- 51 to 77 +/- 57 ml/m2 p < 0.01). Ejection fraction (42 +/- 11 to 48 +/- 13%, p < 0.01) and systolic blood pressure/end-systolic volume (SBP/ESV; 1.06 +/- 0.30 to 1.33 +/- 0.48 mmHg/ml, p < 0.01) were both increased. By radionuclide ventriculography, ejection fraction and peak ejection rate (2.30 +/- 0.74 to 2.80 +/- 0.76 EDV/sec, p < 0.01) were increased, while time to peak ejection, time to peak filling, and peak filling rate were unchanged. Heart rate and double product at exercise were decreased and delta EF was significantly increased (-1.4 +/- 4.1 to 1.6 +/- 4.4%, p < 0.02). The decrease in end-systolic wall stress was consistently related to both the increase in ejection fraction and SBP/ESV, while the decrease in end-diastolic volume was related only to SBP/ESV and not to ejection fraction. Furthermore, there was a direct relationship between the decrease in systolic wall stress and the decrease in end-diastolic volume.(ABSTRACT TRUNCATED AT 250 WORDS)