Scirrhous Cancer Research Articles

Dual blockade of MET and VEGFR2 signaling pathways as a potential therapeutic maneuver for peritoneal carcinomatosis in scirrhous gastric cancer

Scirrhous gastric cancer frequently develops into peritoneal carcinomatosis with malignant ascites, leading to an extremely poor prognosis. We had demonstrated that paracrine hepatocyte growth factor (HGF)-induced MET activation promotes peritoneal carcinomatosis with ascites formation. The vascular endothelial growth factor (VEGF) receptor (VEGFR)/VEGF axis facilitates tumor progression and formation of malignant ascites. This study investigated the role of MET and VEGFR2 in the development of peritoneal carcinomatosis with malignant ascites. Cabozantinib is a dual inhibitor of MET and VEGFR2. We examined the effects of cabozantinib on MET- and VEGFR2-mediated progression of peritoneal carcinomatosis in human scirrhous gastric cancer in vitro and in vivo. Cabozantinib inhibited HGF-stimulated proliferation of scirrhous cancer cell lines NUGC4 and GCIY, with a high potential to generate peritoneal carcinomatosis with ascites fluid, as well as the constitutive proliferation of MKN45 cells with MET amplification. Cabozantinib also inhibited the phosphorylation of both MET and VEGFR2 in scirrhous cancer cells and HGF- or VEGF-stimulated HUVECs. It effectively reduced ascitic fluid and prolonged the survival of NUGC4-inoculated nude mice. In clinical specimens, malignant ascites fluid from patients with peritoneal carcinomatosis contained high levels of HGF and VEGF. Our results strongly suggest that MET- and VEGFR2-mediated signaling pathways play pivotal roles in the pathogenesis of peritoneal carcinomatosis in scirrhous gastric cancer. Thus, the dual blockade of MET and VEGFR2 signaling may be a potential therapeutic maneuver for peritoneal carcinomatosis in scirrhous gastric cancer.

Scirrhous colon cancer presenting as a pseudokidney sign.

This case presents ascending colon cancer with a typical pseudokidney sign on ultrasonography. The cancer extended from the ascending colon to the terminal ileum without intussusception. The pseudokidney sign may represent colon scirrhous cancer.

Efficacy of a Third-Generation Oncolytic Herpes Virus G47Δ in Advanced Stage Models of Human Gastric Cancer

Advanced gastric cancer, especially scirrhous gastric cancer with peritoneal dissemination, remains refractory to conventional therapies. G47Δ, a third-generation oncolytic herpes simplex virus type 1, is an attractive novel therapeutic agent for solid cancer. In this study, we investigated the therapeutic potential of G47Δ for human gastric cancer. In vitro, G47Δ showed good cytopathic effects and replication capabilities in nine human gastric cancer cell lines tested. In vivo, intratumoral inoculations with G47Δ (2 × 105 or 1 × 106 plaque-forming units [PFU]) significantly inhibited the growth of subcutaneous tumors (MKN45, MKN74, and 44As3). To evaluate the efficacy of G47Δ for advanced-stage models of gastric cancer, we generated an orthotopic tumor model and peritoneal dissemination models of human scirrhous gastric cancer (MKN45-luc and 44As3Luc), which have features mimicking intractable scirrhous cancer patients. G47Δ (1 × 106 PFU) was constantly efficacious whether administered intratumorally or intraperitoneally in the clinically relevant models. Notably, G47Δ injected intraperitoneally readily distributed to, and selectively replicated in, disseminated tumors. Furthermore, flow cytometric analyses of tumor-infiltrating cells in subcutaneous tumors revealed that intratumoral G47Δ injections markedly decreased M2 macrophages while increasing M1 macrophages and natural killer (NK) cells. These findings indicate the usefulness of G47Δ for treating human gastric cancer, including scirrhous gastric cancer and the ones in advanced stages.

Abstract 2229: Molecular profile of histological and mutational heterogeneity of adenocarcinoma of the stomach in tumor burden monitoring using circulating tumor DNA

Abstract PURPOSE: This study aimed to perform molecular profiling of the heterogeneous histological and mutational background of primary adenocarcinoma of the stomach for pre- and post-operational tumor burden monitoring using circulating tumor DNA (ctDNA). METHODS: Ten patients with gastric adenocarcinoma who underwent surgical resection for clinical Stage IB or higher were included. Multi-region samples (three sites) per resected specimen were used for cancer cellulality estimation, a 151-gene ClearSeq panel sequencing on Next Generation Sequencer (NGS), and proteomic profiling with 294 proteins using reverse-phase protein arrays (RPPAs). The ctDNA level was evaluated by digital PCR (dPCR) using 25 originally-designed sets with the Hypercool Primer & ProbeTM technology based on mutations of individual tumors. With the median follow-up of 808 days, a total of 187 serial plasma samples were examined for ctDNA. RESULTS: A total of 103 mutations were detected in 30 regions from 10 tumors. Twenty founder mutations (i.e., mutations found in all three regions) were observed in eight tumors whereas two tumors had only non-founder mutations. Twenty-three and 60 non-founder mutations were detected in two regions and one region per tumor, respectively. Variant allele frequencies (VAFs) of founder mutations were higher than those of non-founder mutations (31.2% vs 14.5%; p < 0.01) in primary tumors. With sample regions in which mutations were not detected in trio by NGS, non-founder mutations could be detected by dPCR at low VAFs (ranging from 0.02 to 2.2%) in 95% (19/20) regions. In preoperative patient plasma, ctDNA was detected in 30% (3/10) patients (with a mean VAF of 0.59%). The rate of preoperative ctDNA detection was lower than that of esophageal squamous cell cancer (24/26, 92.3%) and colorectal cancer (10/12, 83.3%) patients. With respect to the tumor stromal effect to ctDNA, no ctDNA was detected in three cases of scirrhous type cancers (0/3, 0%), whereas three of six cases of the other stromal types (3/6, 50%) showed detectable ctDNA (VAF>0.03%). In two relapsed cases with peritoneal dissemination, the elevation of ctDNA was not apparent even at the time of diagnosis of the relapse by CT scan. Among 42 gene-protein matched pairs, the level of proteins did not seem to predict the coding gene mutation. However, tumors with TP53 mutations had significantly higher levels of p53 than those with the wild-type, which was likely due to protein stabilization (p = 0.0004). CONCLUSIONS: Detecting ctDNA in gastric cancer patients may not be as feasible as in other gastrointestinal cancer patients likely due to the heterogeneous histological and mutational background. If applicable, founder mutations are the most suitable marker for detection and monitoring ctDNA. Mutations in the primary tumor itself did not appear to predict protein levels except for TP53. Citation Format: Noriyuki Sasaki, Takeshi Iwaya, Takehiro Chiba, Masashi Fujita, Fumitaka Endo, Mizunori Yaegashi, Ryo Sugimoto, Tamotsu Sugai, Doris Siwak, Lance Liotta, Yilling Lu, Gordon Mills, Hidewaki Nakagawa, Satashi S. Nishizuka. Molecular profile of histological and mutational heterogeneity of adenocarcinoma of the stomach in tumor burden monitoring using circulating tumor DNA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2229.

Effects of 5-fluorouracil, adriamycin and irinotecan on HSC-39, a human scirrhous gastric cancer cell line.

Therapies for patients with scirrhous gastric cancer remain ineffective. Current treatments for gastric cancer based on systemic therapy, such as the combination of S-1 with cisplatin or docetaxel, show good clinical response rates. S-1 plus cisplatin is the standard treatment for HER2-negative advanced scirrhous gastric cancer in Japan. In spite of recent advances in the treatment of gastric cancer, a standard chemotherapy regimen is yet to be established for scirrhous gastric cancer. To develop new therapeutic approaches based on characteristic biological features of cancer cells, we examined the mechanisms underlying the cytotoxicity of anticancer drugs and reactive oxygen species (ROS) toward a human scirrhous cancer cell line, HSC-39, invitro. Anticancer drugs such as 5-fluorouracil (5-FU), adriamycin (ADR)and irinotecan (CPT-11), as well as ROS, were previously shown to have important cytotoxic effects on these tumor cells. We demonstrated that 5-FU effectively induced apoptosis in HSC-39 cells in a dose‑dependent manner, while ADR and CPT-11 induced necrosis and/or aponecrosis. 5-FU effectively inhibited WST-1 decrease in the MTT viability assay, even at low doses where little LDH release was observed, while ADR and CPT-11 only inhibited WST-1 decrease at high doses where LDH release was induced. Moreover, HSC-39 cells showed high sensitivity to H2O2 and NOC-18, but less sensitivity to other ROS, suggesting a link between cell damage and membrane permeability changes induced by H2O2 and NOC-18 or related oxygen radical species such as OH· or ·O2. These results suggest that combination treatment of chemotherapeutics with a fluoropyrimidine such as 5-FU is effective chemotherapy for scirrhous gastric cancer.

Abstract 2368: Differentiated gastric cancer cells have a potential to induce cancer-associated fibroblasts

Abstract Gastric cancer is one of the most common causes of cancer death. Stromal fibroblasts in the tumor microenvironment have recently been implicated in tumor growth, invasion and metastasis. Cancer-associated fibroblasts (CAFs) have been suggested to be responsible for progression of undifferentiated scirrhous gastric cancers with vast fibrous stroma. However, the relationship between differentiated gastric cancer cells and CAFs in tumor progression remains unclear. In this study, we investigated whether differentiated and/or undifferentiated human gastric cancer cells induce CAFs. We used two differentiated human gastric cancer cells (MKN-7, MKN-74) and two undifferentiated human scirrhous gastric cancer cells (KATO-III, NUGC-4). Two types of human normal fibroblasts (WI-38, FEF-3) were used for induction of CAFs. The conditioned media (CM) from all cancer cells were obtained 48 hours after serum starvation. The morphological change and expression of CAF-specific markers (α-smooth muscle actin (α-SMA), fibroblast activation protein-α (FAP)) were analyzed by microscopy and Western blot analysis, respectively, in fibroblasts treated with CM for 120 hours. Moreover, the effect of CM from CAFs in the migration ability of cancer cells was evaluated using in vitro Migration assay. The CM from differentiated MKN-7 and MKN-74 cells induced the morphological change like CAFs in normal human fibroblasts, whereas CM from undifferentiated KATO-III and NUGU-4 cells did not induce. In fact, morphologically changed fibroblasts like CAFs showed increased α-SMA expression, whereas FAP expression was not increased. The CM from morphologically changed fibroblasts, but not normal fibroblasts, significantly enhanced the migration ability of MKN-7 cells. These results suggest that differentiated gastric cancer cells have a potential to induce CAFs in the invasion process of gastric cancers. CAF-targeting therapy may be a promising antitumor strategy in differentiated gastric cancers as well as undifferentiated scirrhous gastric cancers with vast fibrous stroma. Citation Format: Yuncheng Li, Hiroshi Tazawa, Nishizaki Masahiko, Yuuri Hashimoto, Naoto Hori, Ryoichi Katsube, Shinji Kuroda, Kazuhiro Noma, Shunsuke Kagawa, Toshiyoshi Fujiwara. Differentiated gastric cancer cells have a potential to induce cancer-associated fibroblasts. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2368. doi:10.1158/1538-7445.AM2015-2368

Abstract 5338: Preclinical study of telomerase-specific p53 tumor suppressor gene overexpression in human scirrhous gastric cancer cells with different p53 status

Abstract Background: Despite multimodal therapy for scirrhous gastric cancer, many patients show refractory to conventional therapy and rapid progression, leading to poor prognosis. Therefore, the development of novel therapeutic strategy is required to improve the clinical outcome in patients with scirrhous gastric cancers. As novel therapeutic strategies, Ad-p53, a replication-defective adenovirus expressing a tumor suppressor p53 gene, is currently under clinical evaluation for various types of cancers. We recently developed a novel p53-expressing telomerase-specific replication-competent oncolytic adenovirus OBP-702, which induces the p53 gene expression in a telomerase-dependent manner. In this study, we investigated the antitumor effects of Ad-p53 and OBP-702 in human scirrhous gastric cancer cells with different p53 status. Methods: We used 3 human scirrhous gastric cancer cell lines with different p53 status, NUGC4 (p53 wild-type), GCIY (p53 mutant-type), and KATOIII (p53 null). The antitumor effects of Ad-p53 and OBP-702 were evaluated using XTT assay. The 50% inhibiting dose (ID50) value of Ad-p53 and OBP-702 for each cell was calculated using cell viability data obtained 3 days after virus infection. We evaluated the virus-mediated cell death in the sphere-forming cells by LIVE/DEAD assay. The apoptosis- and autophagy-related cell death was assessed by western blot analysis for cleaved poly (ADP-ribose) polymerase (PARP) and p62. Results: Ad-p53 suppressed the cell viability in p53-inactivated scirrhous gastric cancer cells (KATO III, GCIY), whereas p53-intact NUGC4 cells were resistant to Ad-p53. OBP-702 efficiently suppressed the cell viability in all scirrhous gastric cancer cells with different p53 function. The ID50 value of OBP-702 was lower than that of Ad-p53 in all cell lines. LIVE/DEAD assay showed that OBP-702 induced cell death more efficiently than Ad-p53 in the sphere-forming cells. OBP-702 induced more profound p53 expression and apoptotic cell death than Ad-p53. Moreover, only OBP-702 induced not only apoptosis but also autophagy. Conclusions: These results suggest that p53-expressing tumor-specific oncolytic adenovirus OBP-702 is a promising antitumor agent to induce profound cell death in scirrhous gastric cancers with different p53 status. Now, in vivo experiments are under way to investigate the antitumor effect of OBP-702 in the peritoneal dissemination of scirrhous gastric cancer cells. Citation Format: Naoto Hori, Hiroshi Tazawa, Masahiko Nishizaki, Satoru Kikuchi, Shuya Yano, Michihiro Ishida, Megumi Watanabe, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara. Preclinical study of telomerase-specific p53 tumor suppressor gene overexpression in human scirrhous gastric cancer cells with different p53 status. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5338. doi:10.1158/1538-7445.AM2015-5338

Abstract LB-502: Development of three dimensional cancer cell-CAF co-culture system

Abstract Purpose A large number of cell lines have been established and used for cancer research. Monolayer culture of cancer cells alone is a simplified and effective method, however, it does not always reflect the physiological conditions. In this study, we tried to establish a simple co-culture method of cancer cells and CAF (cancer associated fibroblast), to develop three dimensional model of stroma rich tumor with cells such as pancreatic cancer and scirrhous cancer. Method Human tumor tissue samples were obtained from patients who had undergone surgery at the National Cancer Center Hospital East, Japan. Written informed consents were obtained from all patients. BxPC-3 (pancreatic cancer cell line) and A549 (lung cancer cell line) were stained with red-fluorescent dye, PKH-26. Pancreatic cancer derived- and lung cancer derived-CAF were stained with green-fluorescent dye, PKH-67. Cancer cells and CAF are seeded into the microtiter plates for 3D cell culture (NanoCulture® Plate) at a same cell number, and spheroid formation was measured by a fluorescent time lapse microscopy for 3-days. Result BxPC-3 (red) and Pancreatic CAF (green) merged well and formed mosaic spheroids. BxPC-3 (red) and lung CAF (green) did not merge and separately formed spheroids of each cell types. A549 and lung CAF merged, but A549 and Pancreatic CAF did not merge clearly. It is suggested that a specific recognition mechanisms exist by organ types or tumor types, with the adhesion between cancer cell and CAF. A) BxPC-3 and Pancreatic CAF B) BxPC-3 and lung CAF Conclusion In this study, mosaic spheroid were formed by co-culturing of same organ derived cancer cells and CAF. This co-culture system is expected as an effective tool for cancer microenvironment study and/or drug screening targeting with stroma rich tumor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-502. doi:1538-7445.AM2012-LB-502

Primary diagnosis of metastatic breast cancer in the third trimester of pregnancy: A case report and review of the literature

To the best of our knowledge, we are presenting the first documented primary diagnosis of a 32-year-old pregnant patient at 29 + 4 weeks' gestation with poorly differentiated, metastatic scirrhous breast cancer, with negative hormone receptors, HER-2/neu receptor overexpression and metastases in the lumbar spine. The patient was administered neoadjuvant chemotherapy with vinorelbine and trastuzumab, and received ibandronate for the bone metastases. The tumor responded well to treatment; however, treatment was associated with anhydramnios, probably related to the trastuzumab treatment. Delivery was planned for 33 + 5 weeks' gestation by cesarean section due to concurrent breech presentation and anhydramnios, and the infant is in good health. After delivery, the patient underwent a mastectomy. Following completion of six courses of vinorelbine and ongoing treatment with trastuzumab and ibandronate, the patient's tumor went into regression and currently the patient does not present with any clinical evidence of disease.

Co-expression of keratinocyte growth factor and K-sam is an independent prognostic factor in gastric carcinoma

Keratinocyte growth factor (KGF) from gastric fibroblasts have been reported to stimulate proliferation of scirrhous gastric cancer cells with K-samII amplification in a paracrine manner. The aim of this study was to evaluate the clinical significance of the co-expression of K-sam and KGF in gastric carcinomas. A total of 136 primary gastric tumors were investigated by staining with antibodies against K-sam and KGF. K-sam expression on cancer cells and KGF expression on fibroblasts was estimated. The relationship between the K-sam and/or KGF expression and the clinicopathological characteristics were analyzed. K-sam expression was positive in 42 (31%) of 136 gastric carcinomas. K-sam expression was positively correlated with scirrhous cancer (p<0.001), diffuse type (p=0.031), invasion depth (p=0.018) and infiltration type (p<0.001). Prognosis of K-sam positive patients was significantly poorer than that of K-sam negative patients (p<0.001). The prognosis of patients with both K-sam and KGF positive tumors was significantly worse in comparison to either negative tumors (p<0.001). In 94 patients with a curative resection, a multivariate analysis revealed the co-expression of K-sam and KGF to be an independent prognostic factor (p=0.029). In conclusion, the co-expression of K-sam and KGF in gastric cancer might be a useful prognostic factor.

A novel K-samII/FGF-R2 autophosphorylation inhibitor is therapeutically useful for scirrhous gastric carcinoma with K-samII amplification

14070 Background: Scirrhous gastric carcinoma, a diffusely infiltrating also known as linitis plastica-type carcinoma, carries the highest mortality of all gastric cancers. Scirrhous carcinoma cells with amplification of the activated K-samII gene, which encodes fibroblast growth factor receptor type 2 (FGF-R2), have a growth advantage during tumor progression The poor prognosis carried by scirrhous gastric cancer is closely associated with amplification of the K-samII/FGF-R2, a tyrosine kinase growth factor receptor. Ki23057, a newly developed small molecule acting K-samII/FGF-R2 inhibitor, is a kinase inhibitor that competes with ATP for the binding site in the kinase, thus strongly blocking phosphorylation of FGF-R2. The aim of the current study is to clarify the possibility of molecular target therapy with Ki23057 for treating scirrhous gastric cancer. Methods: Five human gastric cancer cell lines were used. OCUM-2MD3 and OCUM-8 were derived from scirrhous carcinomas. MKN-7, MKN-45 and MKN-74 cells were derived from non-scirrhous carcinomas. In vitro effects of Ki23057 on cell growth were determined by calculating the number of cancer cells. The influences of Ki23057 on the MAP kinase and PI3 kinase signaling pathways and the apoptosis pathway in the gastric cancer cells were also examined. For in vivo experiments, the Ki23057 was administered orally to mouse models of peritoneal dissemination. Results: K-samII amplification was found in OCUM-2MD3 and OCUM-8 cells, but not in MKN-7, MKN-45, or MKN-74 cells. Ki23057 significantly inhibited the proliferation of scirrhous cancer cells, but not non- scirrhous gastric carcinoma cells. Ki23057 decreased phosphorylation of K-samII/FGF-R2, ERK and Akt, and increased apoptosis in scirrhous cancer lines. The oral Ki23057 administration significantly (p&lt;0.001) prolonged survival of mice with peritoneal dissemination following injection of OCUM-2MD3 scirrhous cancer cells. Conclusions: A novel K-samII/FGF-R2 phosphorylation inhibitor, Ki23057, appears therapeutically promising in scirrhous gastric carcinoma with K-samII amplification. No significant financial relationships to disclose.

Establishment and characterization of a cell line (NABCA) derived from metastatic lymph nodes of breast scirrhous carcinoma

We successfully established a breast scirrhous carcinoma cell line (designated as NABCA) derived from metastatic tumors of the lymph node. The cells grew as multi-layered cultures without contact inhibition. The population doubling time was approximately 66 h. G-band karyotype of NABCA revealed 66% diploid, XX. Surprisingly, the cells had a number of secretory granules and straight microvilli as a brash border. In heterotransplantation, the cells produced a tumor resembling the original tumor. The NABCA is sensitive to Adriamycin (doxorubicin; KYOWA HAKKO KOGYO, Tokyo, Japan) and Taxol (paclitaxel; Bristol-Myers KK, Tokyo, Japan). This cell line is useful for studying the mechanism of lymphatic metastasis and susceptibility of anticancer drugs in human breast scirrhous cancer.

A Novel Molecular Targeting Compound as K-samII/FGF-R2 Phosphorylation Inhibitor, Ki23057, for Scirrhous Gastric Cancer

Scirrhous gastric carcinoma carries the highest mortality of all gastric cancers. The poor prognosis is reported to be associated with K-samII amplification, which encodes fibroblast growth factor receptor type 2 (FGF-R2). Ki23057, a newly developed small molecule-acting K-samII/FGF-R2 autophosphorylation inhibitor, is a tyrosine kinase inhibitor that competes with adenosine triphosphate for the binding site. The aim of the current study is to clarify the possibility of molecular target therapy with Ki23057 for treating scirrhous gastric cancer. Five human gastric cancer cell lines were used. OCUM-2MD3 and OCUM-8 were derived from scirrhous carcinomas. MKN-7, MKN-45, and MKN-74 cells were derived from nonscirrhous carcinomas. In vitro effects of Ki23057 on cell growth were determined by calculating the number of cancer cells. The influences of Ki23057 on the mitogen-activated protein kinase and phosphatidylinositol 3 kinase signaling pathways and the apoptosis pathway in the gastric cancer cells were also examined. For in vivo experiments, the Ki23057 was administered orally to mouse models of peritoneal dissemination. K-samII amplification was found in OCUM-2MD3 and OCUM-8 cells but not in MKN-7, MKN-45, or MKN-74 cells. Ki23057 significantly inhibited the proliferation of scirrhous cancer cells but not nonscirrhous gastric carcinoma cells. Ki23057 decreased phosphorylation of K-samII/FGF-R2, extracellular signal-regulated kinase, and Akt and increased apoptosis in scirrhous cancer lines. The oral Ki23057 administration significantly (P < .001) prolonged survival of mice with peritoneal dissemination following injection of OCUM-2MD3 scirrhous cancer cells. A novel K-samII/FGF-R2 phosphorylation inhibitor, Ki23057, appears therapeutically promising in scirrhous gastric carcinoma with K-samII amplification.

Clinical application of immunoreactivity of dihydropyrimidine dehydrogenase (DPD) using highly specific antibody against recombinant human DPD (rhDPD) in gastric scirrhous cancer treated with S-1, new DPD inhibitory fluoropyrimidine

14053 Background: Highly specific antibody against recombinant human dihydropyrimidine dehydrogenase (DPD) has been developed to assess immunohistochemically DPD expression in tumors. A new oral DPD inhibitory fluoropyrimidine (DIF), S-1, is reportedly effective against gastric scirrhous cancer. Objective of this study was to assess intra-tumoral levels of DPD immunohistochemically using highly specific anti-DPD polyclonal antibodies, and investigated the relationship between the immunoreactivity of DPD and the anti-tumor effects of S-1 as a DIF in gastric scirrhous cancer patients. Methods: The relationship between immunoreactivity to DPD in biopsy specimens and the effects of chemotherapy was investigated in 61 patients treated with first-line fluoropyrimidine-based chemotherapy (S-1:DIF, 5-FU:non-DIF) for gastric scirrhous cancer. Immunohistochemical staining intensity was semiquantitatively graded (− to 3+) on the basis of the proportion of positively stained cancer cells in the lesions. Results: Total 61 gastric scirrhous cancer patients (M/F: 34/27) were analyzed to date with a median age of 56 (range 30–73). Response rate (partial response) was significantly higher in patients with DPD positive tumors than in those with negative in S-1 group (45.5%, 10.0% : p&lt;0.05), as compared with in 5-FU group (0%, 5.6%: p=0.398). According to median survival time, there was no significant difference between patients with DPD positive tumors (364 days) and those with negative (406 days; p=0.626) in S-1 group or in 5-FU group (181 days and 256 days, respectively; p=0.543). Conclusions: This study indicates that S-1 is effective even in gastric scirrhous cancer with the high level of DPD activity. Further immunohistochemical studies using DIF-based regimens are needed to confirm these results with larger numbers of patients. No significant financial relationships to disclose.

Gastric cancer treated in 1991 in Japan: data analysis of nationwide registry

The Japanese Gastric Cancer Association Registration Committee reported the treatment results and causes of death of patients with primary gastric cancer treated in 1991 at the leading hospitals in Japan. Data of 8851 patients with primary gastric cancer were collected from 113 hospitals, and data of 7935 patients with gastric resection were finally analyzed. The lost-to-follow-up rate was 6.9%; the direct death rate was 1.0%. The cumulative 5-year survival rate (5YSR) of all the patients was 68.2%; 89.9% for Stage I, 69.1% for Stage II, 43.5% for Stage III, and 9.9% for Stage IV. Characteristic findings of the analyzed data were (1) high proportion of early-stage cancer, (2) high resection rate, (3) low mortality rate, (4) low incidence of upper-third cancer, (5) poor treatment results in cases with scirrhous cancer, infiltrating growth, and marked lymphatic or venous invasion, and (6) predominance of systematic (D2) and extended lymphadenectomies possibly resulting in reducing local recurrence and improving survivals.

The death of Napoleon

It was with fascination that I read the paper ‘Channelling the Emperor: what really killed Napoleon?’(August 2004 JRSM1), but I must emphasize that despite its title it is only concerned with the immediate cause of Napoleon’s death. As indicated in my 1996 paper on Napoleon’s health,2 the Emperor first complained of nausea and upper abdominal discomfort after food in July 1820. The dyspepsia worsened to give loss of appetite, constant nausea, frequent vomiting, upper abdominal pain and constipation. There were periods of remission but the pain and vomiting became increasingly insistent. He lost weight and by the end of January 1821 could only manage fluid nourishment, finally taking to his bed on 17 March. On 27 April he vomited blood and his condition deteriorated with further bleeding and lapses into unconsciousness before death on 5 May. At necropsy in front of sixteen observers including seven British doctors, of whom five signed the official report, the external surface of the stomach appeared healthy. On opening, however, it contained altered blood and almost its entire lining was cancerous: a scirrhous cancer had converted the stomach into a leather-bottle stomach with spread to the adjacent lymph nodes and with a perforation that had become sealed off. There was a family history of gastric carcinoma, and Napoleon realized he was dying of the same disease as his father and grandfather, remarking on 15 April, ‘I know the truth and I am resigned’. Clinically the cause of death was clearly gastric carcinoma, but on the publication of my paper in 1996, I received abusive letters with accusations that Napoleon had died from deliberate arsenical poisoning. This theory had received encouragement in 1961 when some, though not all, samples of his hair were found to contain abnormal amounts of arsenic. Arsenic was used in medicaments such as Fowler’s and Donovan’s solutions, and liquor arsenicalis was still in use as a general tonic and for loss of appetite when I qualified. The drawing room at Longwood House in St Helena was redecorated in 1819, but a specimen of the wallpaper applied at that time contained only low levels of arsenic when analysed in 1982. As the immediate cause of death from, say, carcinoma of the prostate may be cardiac failure, so the immediate cause of Napoleon’s death has now been explained to those who still believe he died from arsenic toxicity and not from a carcinoma of the stomach.

Clinicopathologic and Prognostic Characterization of Poorly Differentiated Medullary‐type Gastric Adenocarcinoma

Poorly differentiated medullary-type gastric adenocarcinoma has been reported to differ biologically from poorly differentiated scirrhous adenocarcinoma, and we have characterized the differences in greater detail. Clinicopathologic comparisons were made between the former and latter tumor types as well as well-differentiated gastric adenocarcinoma. Poorly differentiated medullary cancers were smaller, invaded less deeply, and were less likely to arise from the upper part of the stomach than scirrhous cancers. Poorly differentiated medullary cancers less often gave rise to lymph node and peritoneal metastases than scirrhous cancers but more often metastasized to the liver. Venous invasion was more common in medullary than scirrhous or well-differentiated adenocarcinomas. Postoperative survival for patients with poorly differentiated medullary cancers was significantly better than for patients with scirrhous-type cancers but did not differ significantly from survival with well-differentiated adenocarcinoma. The most common cause of death was hematogenous metastasis in patients with medullary or well-differentiated adenocarcinomas, whereas in patients with scirrhous cancer peritoneal metastasis was the most frequent cause. In conclusion, the biologic behavior of poorly differentiated medullary gastric adenocarcinoma was similar to that of well-differentiated adenocarcinoma.

Histopathology of a harbor seal Phoca largha stranded on the coast of Toyama Bay, Sea of Japan

SUMMARY: To estimate the debility and death of a harbor seal Phoca largha stranded on the coast of Toyama Bay, Sea of Japan, on 3 March 1998, histopathological studies on the visceral organs were carried out. Abnormalities were found in four of the organs examined: considerable proliferation of actively phagocytic macrophages in the lymph node; heavy inflammation of the jejunum villi; a benign cortical adenoma in nodules present in the kidney; and a signet ring cell carcinoma (adenocarcinomatous mucocellulare) associated with a marked scirrhous cancer developed in the stomach. The mucocellular carcinoma seemed to have been a main factor in the death of the seal.

Bilateral synchronous spindle cell, apocrine and scirrhous carcinoma breast cancer in a case or malignant lymphoma

An unusual case of synchronous bilateral breast cancer occurring during combination chemotherapy and radiation to the outside of the breast for malignant lymphoma is reported. Two histologically rare carcinomas, spindle cell carcinoma and apocrine carcinoma, were observed in this case. A 77-year-old woman, who had been treated for stage IIIA non-Hodgkin's lymphoma, developed bilateral breast tumors. Aspiration biopsy cytology findings of the tumor in the left breast showed several clusters of adenocarcinoma cells and some large atypical spindle shaped cells, which suggested spindle cell carcinoma. The cytologic findings of the right breast tumor were highly suggestive of scirrhous carcinoma. A modified radical mastectomy was performed on both breasts. Pathological examination disclosed two separate cancer lesions in the left breast. The lesion which had been detected before the operation, was a spindle cell carcinoma. Another lesion, detected for the first time by pathological examination, was an apocrine carcinoma. The lesion in the right breast was a scirrhous carcinoma. Since non-invasive foci were detected in these three cancer lesions, each lesion was thought to be a primary cancer. All dissected bilateral axillary lymph nodes showed malignant lymphoma. Immuno-histochemistry of the spindle cell carcinoma revealed positive immunoreactivity for cytokeratin, which suggested the epithelial as well as mesenchymal nature of this tumor. Synchronous existence of malignant lymphoma and three independent breast cancers including spindle cell carcinoma and apocrine carcinoma is very rare.

Frequent microsatellite instability and loss of heterozygosity in the region including BRCA1 (17q21) in young patients with gastric cancer.

It is known that nearly 5% of gastric carcinomas arise under the age of 40. To elucidate genetic alterations in these patients, we performed studies using microsatellite assay in 27 gastric cancers under 35 years of age, composed of 5 well and 22 poorly differentiated adenocarcinomas. We detected replication errors (RERs) in 18 (67%) of 27 tumors, but no germline mutation in DNA mismatch repair genes (hMLH1 and hMSH2), except fory 3 somatic mutations in the hMLH1 gene. Loss of heterozygosity (LOH) at D17S855, located on chromosome 17q21 (BRCA1), was detected in 8 (40%) of 20 informative cases. In 12 (44%) of 27 cases, LOH on chromosome 17q12-21 including the BRCA1 was found in several neighboring markers in this region, while no mutation was found in the BRCA1 gene. Four (40%) of 10 scirrhous type gastric cancers exhibited wide allelic deletions on chromosome 17q12-21. These results overall suggest that young gastric cancer patients display highly frequent micro-satellite instability that might be due to defect of DNA repair system rather than hMLH1 and hMSH2. In addition, chromosome 17q12-21 including BRCA1 locus may contain a candidate for tumor suppressor gene, particularly in scirrhous type gastric cancers arising in young patients.