Kenichi Yamaguchi

Survivin gene expression positively correlates with proliferative activity of cancer cells in esophageal cancer.

Objective: The correlation between survivin gene expression and the occurrence of spontaneous apoptosis, proliferative activity of cancer cells, tumor angiogenesis, and abnormal p53 nuclear accumulation was evaluated in esophageal cancer. Methods: A total of 57 patients, on whom surgical esophageal resection had been performed between 1993 and 2000, were enrolled in this study. Total RNA was extracted from tumors and noncancerous epithelia. Expression levels of survivin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) messenger RNA (mRNA) were analyzed quantitatively by real-time reverse transcriptase polymerase chain reaction (quantitative PCR). The proliferative activity of cancer cells, apoptotic cancer cells, microvessel density, and abnormal p53 nuclear accumulation were analyzed in these tumors. Results: The expression level of tumor survivin mRNA described as survivin/GAPDH (s/G) ratio was higher than that of noncancerous tissue (p = 0.0003), but did not correlate with lymph node metastasis, with the depth of tumor invasion, with the occurrence of apoptosis or with tumor angiogenesis. However, the tumor s/G ratio positively correlated with the proliferative activity and p53 nuclear accumulation in cancer cells. The 5-year survival rate of 53 patients was 37% (we excluded 4 patients who died from operative complications from this survival study). The 5-year survival rate of 27 patients with a high tumor s/G ratio (28.6%) was lower than that of 26 patients with a low tumor s/G ratio (46.2%, p = 0.041). High tumor s/G ratio was detected as an important prognostic factor independent of tumor stage. Conclusion: Detection of survivin mRNA by quantitative PCR provided us with important prognostic and biological information regarding esophageal cancer patients.

The expression of RCAS1 and tumor infiltrating lymphocytes in patients with T3 gastric carcinoma

Abstract.Background:This study aimed to assess the prognostic value of receptor binding cancer antigen expressed on SiSo cells (RCAS1) expression and host immune response in gastric carcinomas.Methods:We investigated the relationship between RCAS1 expression, density of tumor-infiltrating lymphocytes (TIL), and clinicopathological findings in 129 patients with T3 gastric carcinoma who underwent curative surgery.Results: RCAS1 immunoreactivity was detected in the membrane and cytoplasm of tumor cells. Positive immunoreactivity for RCAS1 was detected in 70 patients (54.3%) and high expression levels of RCAS1 were found in 33 patients (25.6%). The expression of RCAS1 significantly correlated with the histological type of carcinoma and lymph node metastasis. Apoptotic rates in TILs showed marginally higher significance in patients with high RCAS1 expression than in those with low expression. The 5-year survival rates were 77.9% in patients with low RCAS1 expression and 48.4% in those with high RCAS1 expression. Although there was no significant difference in survival between the patients with marked and slight infiltration of TILs, frequent apoptosis in TILs indicated significantly worse prognosis. Patients with low RCAS1 expression survived significantly longer than those with high RCAS1 expression, as did those patients with a high rate of apoptosis in TILs. Multivariate analysis revealed that RCAS1 expression, as well as tumor size and lymph node metastasis, was an independent prognostic factor.Conclusions:The expression of RCAS1 was significantly correlated with poor prognosis in patients with T3 gastric carcinoma. RCAS1 protein may play an important role in the evasion of tumor cells from immunological defense mechanisms in human gastric carcinoma.

Combined resection of invaded organs in patients with T4 gastric carcinoma

Background. To understand the efficacy of gastrectomy combined with the resection of other organs and to refine the indications for this type of surgery, the records of 156 patients with carcinoma of the stomach directly invading adjacent organs or structures (T4 gastric carcinoma) were analyzed retrospectively. Methods. The patients were divided into three groups, as follows: in group A, curative resection was performed by the combined resection of invaded organs or structures; in group B, although combined resection was performed, curative resection could not be performed because of the extent of lymph node metastasis, liver metastasis, and/or peritoneal metastasis; in group C, combined resection was not performed. Results. In patients with peritoneal or liver metastasis, there was no significant difference in prognosis among the three groups. In patients without peritoneal and liver metastasis, the prognosis of group A was significantly better than that of group B or group C, irrespective of the extent of lymph node metastasis or the number of invaded organs. In these group A patients, the 5-year survival rates of those with localized tumors and no lymph node metastasis, those with localized tumors and lymph node metastasis, those with infiltrating tumors and no lymph node metastasis, and those with infiltrating tumors and lymph node metastasis were 100%, 56.2%, 57.1%, and 13.6%, respectively. Conclusions. Combined resection of involved organs should be carried out with curative intent in patients with localized gastric cancer or infiltrating gastric cancer without lymph node metastasis.

Expression of Inducible Nitric Oxide Synthase Is Significantly Correlated with Expression of Vascular Endothelial Growth Factor and Dendritic Cell Infiltration in Patients with Advanced Gastric Carcinoma

Objective: Nitric oxide (NO) is a product of L-arginine to L-citrulline conversion by nitric oxide synthase (NOS). The inducible form of NOS (iNOS) is one of three classes of NOS and the strongest producer of NO. It has been reported that NO correlates with angiogenesis and immune responses in some types of cancer, however, the correlations between iNOS expression, angiogenesis, and immune responses are still unclear in gastric carcinoma. Methods: iNOS expression was determined in 135 gastric cancer patients by immunohistochemical procedures and compared with the expression of vascular endothelial growth factor (VEGF), microvessel (MV) density, and dendritic cell (DC) infiltration to evaluate the effect of iNOS on angiogenesis and immune responses in gastric carcinoma. Results: iNOS expression was detected in 106 (78.5%) of 135 cases. There was a close correlation between iNOS expression and VEGF expression, a correlation with MV density and an inverse correlation with DC infiltration. There was no correlation between iNOS and p53 expression. The prognoses of patients whose tumors expressed iNOS were significantly worse than those of patients whose tumors did not express iNOS. Multivariate analysis indicated iNOS expression was an independent prognostic factor. Conclusion: iNOS might be associated with tumor progression by stimulating angiogenesis and suppressing immune responses in gastric carcinoma.

Expression of the murine double minute gene 2 oncoprotein in esophageal squamous cell carcinoma as a novel marker for lack of response to chemoradiotreatment.

The protein product of the murine double minute gene 2 (MDM2) negatively controls the work of the p53 protein and the retinoblastoma protein (pRB). Recent studies have found that MDM2 expression correlates with chemoresistance of tumor cells. In the present study, the correlation between MDM2 expression and chemoradioresistance was evaluated in patients with esophageal squamous cell carcinoma (ESCC). The immunoreactivities of p53, pRB, and MDM2 were evaluated in 148 surgically resected ESCC by using monoclonal antibodies. The disease-free survival of 107 surviving patients who underwent curative surgery was compared among groups with positive and negative expressions of p53, pRB, and MDM2. High immunoreactivities of p53, pRB, and MDM2 were detected in 47.3%, 64.2%, and 32.4% of cases, respectively. In 107 patients undergoing curative surgery, pRB losses and MDM2 overexpression were found to be poor prognostic factors by univariate analysis. In multivariate analysis, only MDM2 expression was determined to be a prognostic factor independent of the tumor stage. Moreover, we found that MDM2 expression correlates with short survival of patients undergoing postoperative adjuvant chemoradiotherapy. In patients without adjuvant therapy, however, the MDM2 status did not correlate with patient survival. The present results indicate that MDM2 expression may be not only a prognostic marker for patients with ESCC, but also a novel marker for predicting a lack of response to chemoradiotreatment.

Expression of Mcl-1 and p53 proteins predicts the survival of patients with T3 gastric carcinoma

BackgroundThe expression of myeloid cell leukemia 1 (Mcl-1) and p53 proteins was investigated for clinicopathological and prognostic significance in patients with gastric carcinoma.MethodsMcl-1 protein was immunohistochemically examined in 182 patients with gastric carcinoma. The overexpression of p53 was also analyzed in T3 gastric carcinomas.ResultsThe expression of Mcl-1 was detected in 127 (69.8%) patients with gastric carcinoma. Mcl-1 was detected significantly more frequently in the undifferentiated type (P ≪ 0.05) and in the advanced stage of disease (P ≪ 0.05). The prognosis of patients with an Mcl-1-positive tumor was significantly worse than that of those with an Mcl-1-negative tumor (P ≪ 0.05). Multivariate analysis revealed that the expression of Mcl-1 was an independent prognostic factor, as were lymph node metastasis and tumor size. There was no significant relationship between the expression of Mcl-1 and p53. In patients with T3 gastric carcinoma who underwent curative surgery; however, Mcl-1(−)/p53 (−) tumor demonstrated the best postoperative survival rate, whereas Mcl-1(+)/p53(+) tumor had the worst.ConclusionThe expression of Mcl-1 is an indicator of tumor progression and postoperative recurrence in patients with gastric carcinoma. Combined analysis of Mcl-1 and p53 proteins may accurately predict the survival of patients with T3 gastric carcinoma.

Lymph node metastasis of gastric cancer: comparison of Union International Contra Cancer and Japanese systems.

Background:  The pN classification of gastric cancer (GC) in the Japanese system (Japanese Gastric Cancer Association; JGCA) is based on the site and distance of metastatic nodes from the primary tumour. Union International Contra Cancer (UICC) has recently proposed a classification system based on the number of nodes involved (TNM‐1997). The aim of the present study is to assess which classification system is more suitable for providing a prognosis in advanced GC with lymph node metastasis.

Clinicopathological significance of cathepsin D expression in gastric adenocarcinoma.

Objective: An estrogen-regulated lysosomal protease, cathepsin D, has been detected in a variety of tissues. This proteinase has been described as closely associated with tumor progression and metastasis in malignant tumors. The purpose of this study was to determine the clinicopathological and prognostic significance of cathepsin D expression in gastric adenocarcinoma. Methods: In a consecutive series of 478 patients with gastric carcinoma (median follow-up period: 93 months, range: 1–285 months), cathepsin D expression in tumors was quantitatively analyzed with immunohistochemistry using a monoclonal antibody against cathepsin D (clone: 1C11). The percentage of cathepsin-D-positive cancer cells (the CD index) was calculated. In addition, the amount of cathepsin-D-positive stromal cells was evaluated; three grades (high, intermediate, and low) were used for the classification. Results: The mean CD index of 478 tumors was 12.8% (range: 0–100%, median: 8%). The mean CD index of diffuse-type gastric carcinomas (14.9%) was significantly higher than that of intestinal-type carcinomas (10.1%, p < 0.0001). Cathepsin D expression of cancer cells was significantly associated with the depth of tumor invasion in both types. The percentage of tumors with high cathepsin D expression in stromal cells was significantly higher in well-differentiated tumors (25.5%) than in moderately differentiated (12.8%) or in poorly differentiated tumors (19.1%). Cathepsin D expression of stromal cells was significantly associated with the depth of tumor invasion in the intestinal type, in contrast to the diffuse type. Highly expressed cathepsin D in cancer cells was associated with a poor prognosis in both types of carcinoma, but in stromal cells highly expressed cathepsin D was associated to a poor prognosis in the intestinal type only. Conclusion: These results indicate that cathepsin D expression in cancer cells may play an important role in tumor progression in diffuse-type gastric carcinoma, whereas in the intestinal type of carcinoma, cathepsin D expression in stromal cells may play an important role in tumor progression.

Laparoscopic-assisted intraperitoneal chemotherapy for patients with scirrhous gastric cancer.

Background: The prognosis for patients with scirrhous gastric cancer (SGC) is extremely poor. To improve the patients’ prognosis, laparoscopic-assisted intraperitoneal chemotherapy (IPC) was introduced for SGC. In this study, we analyzed whether IPC reduced the number of cancer cells in the peritoneal cavity of patients or changed the gene expression levels of cytokines in the peritoneal cavity. We also investigated whether IPC improved the prognosis of patients with SGC. Methods: Total RNA was extracted from 50 ml of peritoneal wash from 11 SGC patients before and after cisplatin-based IPC. The gene expression levels of survivin, c-myc, transforming growth factor-β (TGF-β), interleukin-2 (IL-2), IL-6, and IL-12 were analyzed using real-time reverse transcription-polymerase chain reaction (RT-PCR) assays. Also, carcinoembrionic antigen (CEA) messenger RNA (mRNA) was used to identify the number of gastric cancer cells in peritoneal washes by the real-time RT-PCR method. The gene expression levels of cytokines and the number of cancer cells in the peritoneal cavity were compared before and after cisplatin-based IPC treatment. Results: Before IPC, the gene expression of IL-2 from peritoneal washes of patients was significantly suppressed compared to the controls (p = 0.029); however, other gene expression levels did not differ. In 7 cases, more than 90% of the cancer cells were removed from the peritoneal cavity after cisplatin-based IPC. These 7 cases were named the IPC effective group, and the remaining 4 cases were named the IPC ineffective group. In the IPC effective group, elevated IL-2 and IL-6 genes were detected in 5 (71%) and in 6 (86%) after IPC. The correlation between IPC effectiveness and elevated gene expression after IPC (IL-2: p = 0.137, and IL-6: p = 0.044) was observed. However, the 50% survival period of the IPC effective group (9 months) was not different from that of that of the IPC ineffective group (6 months, p = 0.267). Conclusion: IPC effectiveness may correlate with elevation of gene expression of inflammatory cytokines, such as IL-2 and IL-6 in the peritoneal cavity after IPC. However, the prognostic benefits of IPC for SGC patients remain unclear.

Quantitative Analysis of Heparanase Gene Expression in Esophageal Squamous Cell Carcinoma

Background: Heparan sulfate proteoglycans, the main components of the extracellular matrix, are recognized as important components of signal transduction and play an important role in tumor progression. Heparanase (hep) degrades heparan sulfate proteoglycans, but the clinical importance of hep is unclear. In this study, we investigated the clinicopathologic importance of hep messenger RNA (mRNA) expression in esophageal squamous cell carcinoma (ESCC).Methods: Fresh tumors and noncancerous epithelia were obtained from 57 ESCC patients after esophagectomy. Expression levels of hep and glyceraldehyde-3-phosphate dehydrogenase mRNA were quantitatively analyzed by real-time reverse transcriptase-polymerase chain reaction. Apoptotic cancer cells and microvessel density were evaluated immunohistochemically.Results: The relative hep mRNA expression level (hep:glyceraldehyde-3-phosphate dehydrogenase ratio) in ESCC was lower than in noncancerous tissue (P < .001). Tumor hep expression decreased according to tumor progression and correlated with the occurrence of apoptotic cancer cells, but not with tumor microvessel density. Moreover, low hep expression correlated with poor patient survival.Conclusions: Reduced hep mRNA expression might result in abnormal cell growth and correlate with ESCC progression.