Up-regulation of histamine H4 receptors contributes to splenic apoptosis in septic mice: counteraction of the antiapoptotic action of nuclear factor-kappaB.

The histamine H(4) receptor is the most recently identified receptor and is considered to play a role in a variety of inflammatory diseases. Histamine levels in the plasma are known to be elevated in animal models of sepsis and in septic patients. The aim of this study was to test the hypothesis that the H(4) receptor may play a significant role in the pathophysiology of sepsis. Polymicrobial sepsis was induced by cecal ligation and puncture in BALB/c mice. Although the H(4) receptor gene was undetectable in normal peripheral key organs, with the exception of the spleen, the expression levels of this gene were highly up-regulated in all those organs of septic mice. In vivo transfection of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide, but not of its scrambled form, resulted in a great inhibition of sepsis-induced overexpression of the H(4) receptor gene. In septic mice, marked increases in caspase-3 activation and follicular lymphocyte apoptosis in spleens were strongly suppressed by systemic treatment with synthetic small interfering RNA (siRNA) targeted to the H(4) receptor. This was associated with the up-regulation of a number of antiapoptotic proteins. These antiapoptotic effects of H(4) receptor siRNA treatment were all inhibited by further application of NF-kappaB decoy oligonucleotide. Our results suggest that superinduction of the histamine H(4) receptor gene in peripheral key organs, including the spleen, that is promoted by sepsis is transcriptionally controlled by NF-kappaB, whereas stimulation of this receptor is involved in the development of sepsis-induced splenic apoptosis through counteraction of the antiapoptotic action of NF-kappaB.

Impact of loss of heterozygosity of encoding phosphate and tensin homolog on the prognosis of gastric cancer.

BACKGROUND AND AIM: Encoding phosphate and tensin homolog (PTEN) is a cancer suppressor gene and it has been assumed that gene mutation and loss of heterozygosity (LOH) occurs frequently in various types of carcinoma. However, the role of LOH of PTEN and its outcome variables in gastric cancer have not been well established. In the present study, we investigated the roles of PTEN, LOH and their outcomes. METHODS: Fresh frozen tumor samples from 119 gastric cancer patients with a primary diagnosis of gastric carcinoma were evaluated for LOH of PTEN using an automated sequencer. Results were compared with pathological parameters. The median follow-up period was 559 days. RESULTS: Loss of heterozygosity of PTEN was observed in 17.1% of patients (13/76) diagnosed with gastric cancer. No particular relationship was found with any clinicopathological factor. However, the prognosis of patients with LOH of PTEN was significantly poor. Multivariate analyses revealed that vascular invasion, invasion depth, LOH of PTEN, histology and lymph node metastasis were correlated with survival of the patient. CONCLUSIONS: Even though mutation of PTEN in gastric cancer has rarely been reported, according to our findings, LOH of PTEN frequently occurs in gastric cancers and is correlated with disease-related deaths. The LOH of PTEN is an independent prognostic factor and PTEN is a candidate as a haploinsufficient tumor suppressor in gastric cancers.

Akt is frequently activated in HER2/neu-positive breast cancers and associated with poor prognosis among hormone-treated patients.

Akt/PKB is a serine/threonine kinase that plays an important role in survival when cells are exposed to different apoptotic stimuli. Aberrant activation of Akt/PKB in breast carcinoma is associated with poor prognosis and resistance to endocrine therapy and chemotherapy. The Akt signaling pathway currently attracts considerable attention as a new target for effective therapeutic strategies. We therefore investigated the relationship between activation of Akt and clinicopathologic variables including hormone receptor and HER2/neu status. Breast cancer tissues obtained from 252 patients were utilized for this study. We evaluated Akt activation by immunohistochemical assessment of the expression of phosphorylated Akt (pAkt) at Ser-473. Eighty-four cases (33.3%) were diagnosed as positive for pAkt expression. pAkt was significantly associated with HER2/neu overexpression (p < 0.0001). There was an inverse correlation between pAkt and PR expression (p = 0.0321); however, there was no association between pAkt and ER expression. Survival analysis showed that pAkt positivity was associated with poor disease-free survival in cases with postoperative hormone therapy; however, there was no association in cases without hormone therapy. Our results indicate that Akt activation induced poor prognosis in patients who received adjuvant hormone therapy. This finding suggests that inhibition of the Akt signaling pathway may increase the efficacy of hormone therapy and improve the prognosis of patients who receive adjuvant hormone therapy.

Genetic mutual relationship between PTEN and p53 in gastric cancer.

Both PTEN (encoding phosphate and tensin homologue) and p53 are known as cancer suppressor genes, and they are assumed that their gene mutations and loss of heterozygosity (LOH) occur frequently in various types of carcinoma. In the present study, we investigated both the p53 mutation and LOH of PTEN in 113 gastric cancer patients. We observed the LOH of PTEN in 11.1% of the patients with normal p53s and 46.2% of the patients with p53 gene mutations. The result that LOH of PTEN was frequently observed in the cases with p53 gene mutations and other data in this study suggested that both PTEN and p53 have complimentary roles in gastric carcinoma development.

Akt phosphorylation associates with LOH of PTEN and leads to chemoresistance for gastric cancer.

Growth factor receptor-mediated signal transduction has been implicated in conferring resistance to conventional chemotherapy on cancer cells. We describe a pathway that involves AKT/PI3K to mediate chemoresistance in gastric cancer patients. Primary gastric carcinoma tissues and corresponding normal mucosa were obtained from 76 gastric cancer patients who underwent surgery in the Department of Surgery II in Kyushu University Hospital from the years 1996-2000. AKT activation was investigated by immunostaining with a phosphorylation-specific antibody, and LOH (loss of heterozygosity) of PTEN was studied in the same samples. AKT was phosphorylated in 22 cases (28.9%) of gastric cancer cases. AKT and phosphorylated AKT were not correlated with any clinicopathological factor. We found that the gastric cancer patients who had higher AKT phosphorylation (activated AKT) seemed to have LOH of PTEN (p = 0.0008). When the chemotherapeutic sensibilities of these patients were studied in an MTT assay, it was found that the activated AKT was associated with increased resistance to multiple chemotherapeutic agents (5-fluorouracil, adriamycin, mitomycin C and cis-platinum). The results of our study indicate that AKT activation and LOH of PTEN plays an important role in conferring a broad-spectrum chemoresistance in gastric cancer patients. It also indicates that AKT may therefore be a novel molecular target for therapies or chemosensitivity tests that improve the outcomes of gastric cancer patients.

Suppression of the phytohemagglutinin response to lymphocytes is an independent prognosticator in patients with squamous cell carcinoma of the esophagus.

BACKGROUND: The preoperative immunological condition of patients with malignant tumors should be considered in determining the prognosis. METHODS: A lymphoblastic transformation test in which lymphocytes were stimulated with phytohemagglutinin (PHA) was performed for 155 patients with esophageal squamous cell carcinoma (SCC). RESULTS: Multivariate analysis demonstrated that a lower preoperative PHA response (p = 0.028), as well as the depth of the tumor (p = 0.011), lymph node metastasis (p = 0.0003), lymphatic permeation (p = 0.002), and the incidence of postoperative complication (p = 0.016) were independent factors associated with a poor prognosis for patients with SCC of the esophagus. CONCLUSIONS: A suppressed PHA response is an additional significant prognosticator for SCC of the esophagus.

Immunohistochemical expression of C-reactive protein in squamous cell carcinoma of the esophagus - significance as a tumor marker.

C-reactive protein (CRP) is a prototype acute phase protein which has been known to be synthesized in hepatocytes. Although serum elevation of CRP has been reported to be an indicator of the unfavorable outcome of the patients in some malignant tumors, the expression of the protein in carcinoma cells has not been investigated. The aim of the current study was to assess the immunohistochemical expression of CRP in squamous cell carcinoma (SCC) of the esophagus and to find its biological significance. Immunohistochemical examination for CRP expression was performed for 37 advanced esophageal SCCs with the depth of T2, T3 or T4, which had been surgically resected without preoperative therapy for the patients. Eighteen carcinomas (48.6%) demonstrated immunohistochemical CRP expression. Univariate analysis showed that the prognosis of the patients with esophageal SCCs expressing CRP was significantly worse than that in patients with tumors without CRP expression (P=0.017). Moreover, CRP expression was found to be an independent prognosticator in patients with esophageal SCCs in the multivariate analysis (P=0.036). To the best of authors' knowledge, this is the first report that demonstrated the possible carcinoma-related expression of CRP in SCCs of the esophagus and its biological significance as the prognosticator of the patients.

Cyclin A expression in superficial squamous cell carcinoma of the esophagus and coexisting infiltrated lymphocyte follicle.

Cyclin A is a protein kinase to act a pivotal role in the mitotic phase of the cell cycle. The purpose of the current study was to elucidate the biological significance of immunohistochemical expression of cyclin A in superficial squamous cell carcinoma (SCC) of the esophagus. Immunohistochemical staining of cyclin A was performed for 45 samples of esophageal superficial SCCs. Clinicopathological features were compared between SCCs with and without cyclin A expression. Twenty-five superficial SCCs (55.6%) had positive expression of cyclin A and the other 20 (44.4%) did not. No significant difference regarding clinicopathological characteristics between esophageal SCCs with and without cyclin A expression. Infiltration of lymphocytes with germinal center cells was observed beneath 17 (68.0%) out of 25 superficial SCCs with cyclin A expression and 15 (75.0%) out of 20 superficial SCCs without cyclin A expression. Although 16 (94.1%) out of 17 superficial SCCs with cyclin A expression were associated with cyclin A expression in germinal center cells in infiltrated lymphoid follicles beneath the tumors, only 2 (13.3%) out of 15 superficial SCCs without cyclin A expression coexisted with cyclin A expression in lymphoid follicles beneath the tumors (P<0.0001). Cyclin A expression in the germinal center cells of the lymphoid follicles beneath the superficial SCCs of the esophagus might be an immunological signal toward the proliferation and progression of the tumors.

The relation of alcohol consumption and cigarette smoking to the multiple occurrence of esophageal dysplasia and squamous cell carcinoma.

BACKGROUND: The unique pathologic features of esophageal tumors in patients with esophageal cancer includes the presence of multiple occurrence within the esophagus. The aim of this study is to clarify the molecular mechanism of carcinogenesis of multiple esophageal squamous cell carcinomas in the Japanese. METHODS: We studied the relationship between the incidence of patients with multiple carcinomas and the coexistence of dysplasia lesions with p53 protein accumulation, alcohol consumption, and cigarette smoking. Among 76 cancer lesions and 60 cases of dysplasia, p53 accumulation was studied by means of immunohistochemical analysis. RESULTS: The incidence of patients with multiple carcinomas in the high-risk group was 33%, and the incidence of patients with a coexistence of dysplasia in the high-risk group was 67%. The incidence of patients with multiple carcinomas or the coexistence of dysplasia in the high-risk group was much higher than that of the middle-risk and low-risk groups (P <.0001 and P =.04, respectively). The average number of abnormal epitheliums, such as cancer and dysplasia, in the high-risk group was 3.2 +/- 2.1. The average number of abnormal epitheliums was much higher than that of the other groups (P =.02). For carcinoma lesions, the incidence of lesions with a positive p53 protein accumulation in the high-risk group was 91%. Regarding dysplasia lesions, the incidence of lesions with a positive p53 protein accumulation in the high-risk group was 80%. The incidence of both cancer and dysplasia lesions with a positive p53 protein accumulation in the high-risk group was higher than that of the other groups. CONCLUSIONS: The pattern of p53 accumulation in dysplasia in the high-risk group was closely similar to that in cancer of the high-risk group. These findings support the concept of field carcinogenesis of the esophagus.

Surgical and oncological advances in the treatment of esophageal cancer.

BACKGROUND: Chemoradiotherapy (CR) and hyperthermochemoradiotherapy (HCR) have been performed on numerous patients with esophageal cancer. These neoadjuvant therapies for esophageal cancer are done widely. The purpose of this study is to demonstrate the recent advances in surgical and oncological treatment. METHODS: From 1967 to 2000, 847 patients who underwent an esophagectomy were classified into 4 groups according to the date of operation. Group 1 consisted of 110 patients who underwent an esophagectomy in the first 10-year period (1967-1976), group 2 consisted of 194 patients who had operations from 1977 to 1986, group 3 comprised 400 patients who had operations from 1987 to 1996, and group 4 comprised 143 patients who had operations from 1997 to 2000. From 1967 to 2000, 322 patients with neoadjuvant therapy and esophagectomy were classified into 6 groups according to the kinds of anticancer drugs that were administered. Group A received regimen A, using BLM (5 mg iv) 6 times as the chemotherapeutic drug in the early period (1965-1990); group B received regimen B, using cis-diaminedichloroplatinum (CDDP) (40 mg/m2) 3 times as the chemotherapeutic drug in the second period (1990-1997); and group C received regimen C, using CDDP (40 mg/m2) and 5FU (250 mg/m2) daily in the most recent period (1997-2000). The HCR group was also divided into the following 3 groups: Group D, who received regimen A and hyperthermia 6 times in the early period; group E, who received regimen B and hyperthermia 6 times in the next period; and group F, who received regimen C and hyperthermia 6 times in the most recent period. The local response and the long-term results were investigated. RESULTS: A complete removal of the primary tumor was achieved in 29%, 39%, 62%, and 68% of the patients in groups 1, 2, 3, and 4, respectively. The 30-day operative mortality rates were 11%, 4%, 1%, and 0% in groups 1, 2, 3, and 4, respectively. The 5-year survival rates for all patients in groups 1, 2, and 3 were 16.7%, 19.2%, and 44.4%, respectively. The cases in which CR or HCR was evaluated to be effective numbered 44 (48.4%) in group A, 22 (73.3%) in group B, 8 (66.7%) in group C, 79 (63.7%) in group D, 36 (73.5%) in group E, and 12 (75.0%) in group F. Our clinical results thus showed CDDP to have a greater effect than BLM, while HCR was shown to have a greater effect than CR. CONCLUSIONS: Preoperative therapy, especially using CDDP and hyperthermia, has improved thanks to recent advances in the treatment of esophageal cancer.

[Evaluation of multi-modality treatment for the patients with advanced esophageal cancer].

Hyperthermia has been proved to be intensely cytotoxic to malignant cells when combined with anticancer agents and irradiation. We have applied hyperthermo-chemo-radio-therapy (HCR) using a radiofrequency system with an endotract electrode to the patients with advanced esophageal cancer. In this study, 132 patients with advanced esophageal cancer with invasion to the neighboring structures, who underwent non-curative operation, were analysed. The markedly effective cases was observed in 12.2% in HCR group, while in 2.5% in chemoradiotherapy (CR) group, microscopically. The survival rate of the HCR group was significantly better than that of the CR group (p < 0.05). Pre- and post-operative leukocytopenia caused by preoperative treatment was not found to be a risk factor of postoperative complication. It is important to continue to clarify the factors influencing the effectiveness of preoperative HCR in the future. We hope that HCR will play an even more important role in treatment of esophageal cancer.

Role of dihydropyrimidine dehydrogenase activity in patients with esophageal cancer.

Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme involved in the degradation of 5-FU. DPD activity in peripheral blood mononuclear cells of 30 esophageal cancer patients treated with 5-FU and low-dose CDDP with irradiation was determined at the beginning of each cytostatic cycle, the objective being to determine if DPD activity is related to the occurrence of side-effects and responses to therapy. The DPD activity showed interpatient variability (mean: 325.5 pmol/min/mg protein). 5-FU-related side-effects tended to be registered more frequently in patients with low DPD activity. In particular, nausea occurred in 30.8% of patients in the high DPD activity group but, in 70.6% in those with low DPD activity (p < 0.05). The relationship between the histological response to therapy and DPD activity was nil. We propose that determination of DPD activity prior to initiation of 5-FU-based chemotherapy for patients with esophageal cancer could aid in identifying those at risk for toxicity.