A case of primary signet-ring cell carcinoma of the cervix containing full genome of human papillomavirus 16.

We herein present a case of primary signet-ring cell carcinoma of the cervix. Pelvic magnetic resonance imaging revealed a 38-mm cervical tumor, and computed tomography revealed no findings suggestive of distal metastasis or other tumor origins. Gastrointestinal endoscopy showed no abnormal findings. Histopathology revealed signet-ring cell-type mucinous adenocarcinoma. By immunohistochemistry, tumor cells were negative for the mammary neoplasm marker, gross cystic disease fluid protein 15 and gastrointestinal neoplasm markers, MUC2, MUC6, and CDX2, but positive for p16. These findings suggested human papillomavirus (HPV)-related adenocarcinoma of the cervix. HPV genotyping assays with exfoliated cervical cells and formalin-fixed paraffin-embedded tissues demonstrated HPV16 positivity, suggesting that the primary origin of the tumor was the cervix. The full HPV16 genome was amplified by polymerase chain reaction from exfoliated cervical cells, and the full-genome sequence was determined by next-generation sequencing. This is the first report of primary signet-ring cell carcinoma of the cervix containing the full HPV16 genome.

Genetic analysis of a case of Helicobacter pylori-uninfected intramucosal gastric cancer in a family with hereditary diffuse gastric cancer.

Germline mutations in CDH1, encoding E-cadherin, are known to be the causative mechanism of hereditary diffuse gastric cancer (HDGC). We encountered two cases of gastric cancer in a Japanese family with HDGC. A 28-year-old man (Case 1) died of advanced gastric cancer. His younger sister aged 27 (Case 2) was diagnosed with intramucosal signet ring cell carcinoma (SRCC). Both had identical germline CDH1 mutations, but Case 1 was positive for Helicobacter pylori infection, whereas Case 2 was negative. Case 2 underwent total gastrectomy. Whole-exome sequencing of an intramucosal SRCC in Case 2 revealed seven somatic mutations including one in CDH1. The six non-CDH1 mutations were classified as non-driver mutations. Decreased expression of E-cadherin in intramucosal SRCC was confirmed by immunohistochemistry. Our report demonstrated that CDH1 mutation was the only active driver mutation in Helicobacter pylori-uninfected intramucosal SRCC.

Cervical Carcinoma With Divergent Neuroendocrine and Gastrointestinal Differentiation.

Neuroendocrine carcinomas of the uterine cervix are rare tumors with aggressive behavior. They comprise <4% of cervical carcinomas. They may coexist with both adenocarcinoma and squamous cell carcinoma of cervix. Signet ring carcinoma of cervix is a rarer entity and less than 20 cases have been described in the literature. We present a case of a 34-year-old female who presented with systemic thrombosis, splenic mass and a cervical mass which on biopsy showed divergent differentiation of primitive large cell neuroendocrine carcinoma with signet ring cells. The cervical tumor was positive for human papilloma virus 16/18 by in situ hybridization, confirming cervical origin of the tumor. This unusual presentation and morphology needs to be recognized and appropriately evaluated when patients present with tumors of unknown origin in metastatic sites.

Determinants of Epstein-Barr virus-positive gastric cancer: an international pooled analysis.

BACKGROUND: Meta-analyses of the published literature indicate that about 9% of gastric cancers contain Epstein-Barr virus (EBV), with consistent and significant differences by sex and anatomic subsite. This study aimed to identify additional determinants of EBV positivity and their joint effects. METHODS: From 15 international populations with consistent laboratory testing for EBV, we pooled individual-level data for 5081 gastric cancer cases including information on age, sex, subsite, histologic type, diagnostic stage, geographic region, and period of diagnosis. First, we combined population-specific EBV prevalence estimates using random effects meta-analysis. We then aggregated individual-level data to estimate odds ratios of EBV positivity in relation to all variables, accounting for within-population clustering. RESULTS: In unadjusted analyses, EBV positivity was significantly higher in males, young subjects, non-antral subsites, diffuse-type histology, and in studies from the Americas. Multivariable analyses confirmed significant associations with histology and region. Sex interacted with age (P=0.003) and subsite (P=0.002) such that male predominance decreased with age for both subsites. The positivity of EBV was not significantly associated with either stage or time period. CONCLUSION: Aggregating individual-level data provides additional information over meta-analyses. Distinguishing histologic and geographic features as well as interactions among age, sex, and subsite further support classification of EBV-associated gastric cancer as a distinct aetiologic entity.

Increased risk of tuberculosis after gastrectomy and chemotherapy in gastric cancer: a 7-year cohort study.

BACKGROUND: Gastrectomy for peptic ulcers and chemotherapy for malignancy are known risk factors for tuberculosis (TB). However, this relationship has rarely been investigated in patients with gastric cancer. METHODS: In a retrospective cohort study from 2000 to 2006, data for 2215 patients diagnosed with gastric cancer at our hospital were compared with data from the Centers for Disease Control (CDC), Taiwan, to identify confirmed cases of TB. RESULTS: In patients with gastric cancer without a history of gastrectomy and without previous anti-TB treatment, the overall crude incidence of new-onset TB was 788 per 100,000 person-years. Compared with the general population, the overall age-sex standardized incidence (SI) in gastric cancer patients was 134.3 (SI ratio [SIR]: 2.11, p < 0.05), and the recurrence rate among patients with previous anti-TB treatment was 18% (4/22) after gastric cancer diagnosis. Gastrectomy was a significant risk factor for active TB incidence [SI 159 (95% confidence interval [CI], 80-237, SIR: 2.5, p < 0.05), and chemotherapy alone seemed to be a risk factor, but was without statistical significance (SIR: 12.5, p > 0.05). Multivariate analysis showed old age, male gender, previous anti-TB treatment, and gastrectomy as significant risk factors for TB. In stratified analysis, an interaction between old TB patterns on chest films and chemotherapy was observed. CONCLUSIONS: Old age, male gender, previous anti-TB treatment, and gastrectomy were significant risk factors for TB. An increased risk of TB incidence after chemotherapy was observed in patients with old TB pattern chest films initially, suggesting an interaction between chest film patterns and chemotherapy.

Epstein-Barr virus-associated gastric carcinoma among patients with pernicious anemia.

Approximately 9% of gastric carcinomas worldwide are associated with Epstein-Barr virus (EBV), making it the most frequent EBV-associated malignancy. Pernicious anemia, a condition with chronic gastritis and achlorhydria, is strongly associated with gastric carcinoma. Both chronic inflammation and the lack of stomach acid may influence the likelihood of EBV infection of the neoplastic gastric epithelium, but the prevalence of EBV-associated gastric carcinoma among patients with pernicious anemia is unknown. Therefore, we conducted a Danish nationwide case-control study comparing gastric carcinoma patients with pernicious anemia (PA-GC) with those without pernicious anemia (nonPA-GC), frequency matched 1:2. Tumor tissues were reclassified by expert histopathologists blinded to pernicious anemia and EBV status. In total, 186 samples (55 PA-GC and 131 nonPA-GC) were identified. EBV-associated gastric carcinoma (EBV-GC) was more common among PA-GC compared with nonPA-GC, adjusted odds ratio (OR) = 2.53 (CI: 0.88; 7.14), p = 0.08, with further adjustment for lymphocytic infiltrate OR = 2.94 (0.99-8.67), p = 0.05. Gastric carcinomas with signet-ring cell morphology were significantly less common in patients with PA-GC compared with nonPA-GC (OR = 0.05, CI 0.01; 0.24). Although these conditions are rare, we found suggestive evidence that EBV-associated gastric carcinomas are more common among gastric carcinoma patients with pernicious anemia compared with those without.

Primary signet-ring cell carcinoma of the cervix: case report and review of the literature.

Mucinous adenocarcinoma of the cervix has 5 subtypes: endocervical, intestinal, signet-ring cell, minimal deviation, and villoglandular. There are only rare reports of primary signet-ring cell carcinoma of the cervix in the literature. Herein we report a 53-year-old woman with cervical adenocarcinoma with signet-ring cell morphology. Thorough systemic examination did not reveal another primary focus. DNA extraction from paraffin-embedded tissue revealed the presence of human papilloma virus (HPV) type 18, which supports the cervical origin of the tumor. Signet-ring cell morphology can be observed in both benign and malignant lesions of the uterine cervix. Most of the malignant lesions are metastatic. Histological features and immunohistochemical profiles are discussed, and a review of signet-ring cell morphology in the uterine cervix is included.

[CD21-independent infection of a human signet ring cell gastric carcinoma cell line by Epstein-Barr virus].

OBJECTIVE: To understand Epstein-Barr virus (EBV) infection of gastric carcinoma cells. METHODS: The authors tested the infection of a signet ring cell line HSC-39 derived from human gastric carcinoma with Akata and P3HR-1 strains of EBV. Akata and P3HR-1 infected of EBV cell clones were isolated by a limiting dilution method. RESULTS: EBV-encoded small RNAs (EBERs) were expressed in the infected cells with each EBV strain by in situ hybridization. The EBV infected parental cells and most clones expressed EBNA1, but not EBNA2, latent membrane protein (LMP) 1 and LMP2A. Both EBV strains infected parental cells and clones presented type I latency. The uninfected HSC-39 cells were negative for CD21 expression; however, the Akata but not P3HR-1-infected clones were positive for CD21 expression at mRNA level. CONCLUSION: These results demonstrated that EBV infecting HSC-39 by CD21-independent pathway. This study also defined a signet ring cell line as a new target for EBV.

[Expression of Epstein-Barr virus genes in EBV-associated gastric carcinoma].

BACKGROUND & OBJECTIVE: Epstein-Barr virus (EBV) is associated with the development of many malignant tumors. The forms of EBV and the expression of EBV genes in Burkitt's lymphoma and nasopharyngeal carcinoma (NPC) have been reported. These studies showed that the forms of EBV and the expression of EBV genes are various in different types of malignancies. However, there were only a few reports about the expression of EBV genes, especially the lytic genes, in gastric carcinoma tissues. This study was to determine the expression of EBV latent and lytic infection genes in gastric carcinoma at the transcriptional level by RT-PCR and Southern hybridization,and investigate the relationship between EBV-encoded genes and the tumorigenesis of gastric carcinoma at the molecular level. METHODS: One hundred and eighty-five gastric carcinoma and corresponding para-carcinoma tissues were tested for EBV genome by polymerase chain reaction (PCR)-Southern analysis. EBV-encoded small RNA 1 (EBER1) of the PCR positive specimens was determined by in situ hybridization (ISH). Gastric carcinoma with positive EBER1 signals was confirmed EBV- associated gastric carcinoma (EBVaGC). RT-PCR and Southern hybridization were used to determine the expression of nuclear antigen (EBNA) promoters (Qp, Wp and Cp), EBNA 1 and 2,latent membrane protein (LMP) 1, 2A, and 2B and lytic genes (immediate-early genes BZLF1 and BRLF1, early genes BARF1 and BHRF1, late genes BcLF1 and BLLF1) in EBVaGCs. RESULTS: There were 13 EBV positive samples in gastric carcinomas (7.03%), but no EBV positive sample in corresponding para-carcinomas. The transcripts of Qp were detected in all of the 13 EBVaGCs tissues, while both Wp and Cp were silent. All of the 13 cases expressed EBNA1 mRNA, but no EBNA2, LMP1, and LMP2B mRNA. LMP2A mRNA was detected in 5 of the 13 cases. Of the 13 EBVaGCs, 7 exhibited BcLF1 transcript and 2 exhibited BHRF1 transcript. The transcripts of BZLF1 were detected in 6 cases, and those of BARF1 also in 6 cases. No BLLF1 and BRLF mRNA were detected in the 13 EBVaGCs. CONCLUSIONS: The latent pattern of EBV in EBVaGCs corresponds to the latency I or unique latency I/II, intermediate between the latency I and II. Part of lytic infection genes are expressed in EBVaGCs tissues. BARF1 and BHRF1 genes express in part of gastric carcinomas and their roles in gastric tumorigenesis need to be further studied.

Characterization of Epstein-Barr virus infection in a human signet ring cell gastric carcinoma cell line, HSC-39.

In order to study the mechanism of Epstein-Barr virus (EBV) infection in gastric carcinoma cells, we characterized the EBV infection in signet ring cell line HSC-39, derived from a human gastric carcinoma. HSC-39 cells were highly susceptible to cell-free EBV infection by Akata and P3HR-1 EBV strains. EBV nuclear antigen (EBNA) and EBV-encoded small RNA (EBER) were detected in the infected cells. Akata and P3HR-1 EBV-infected cell clones were isolated by a limiting dilution technique. The Akata and P3HR-1 EBV-infected clones differed from each other in morphology and growth patterns. Akata EBV-infected clones had lower growth rates than did P3HR-1 EBV-infected clones in both liquid and soft agar mediums. Both the infected HSC-39 cells and the clones expressed EBNA1 and EBER, but did not express EBNA2, latent membrane protein (LMP) 1 and LMP2A. The Q promoter (p), but not the Cp/Wp for EBNA transcription, was active in the infected HSC-39 cells and all clones. No lytic infection was observed in either infected parental cells or any clones. Uninfected HSC-39 cells did not express a principal EBV receptor CD21; however, Akata but not P3HR-1 EBV-infected clones expressed low levels of CD21 mRNA. These results demonstrate that the cellular phenotypes of HSC-39 cells are altered by EBV infection in strain-specific manner. We propose the HSC-39 cell line as a model target for the study of the mechanism and significance of EBV infection in gastric carcinoma.

Detection of the Epstein-Barr virus in primary adenocarcinoma of the lung with Signet-ring cells.

The Epstein-Barr virus (EBV) is directly implicated in the pathogenesis of a variety of undifferentiated carcinomas and has also been identified in conventional adenocarcinomas of the stomach. To date, the association of EBV with non-small cell lung carcinoma is restricted to Asian patients. To evaluate the presence of EBV in lung cancers from Europeans, we investigated primary lung adenocarcinomas with a similar morphological tumour pattern to those of the stomach, specifically rare tumours with components of signet-ring cells. Three tumours of signet-ring cell type were examined by means of polymerase chain reaction (PCR). To localise the virus to the neoplastic cells, in situ hybridisation (ISH) was performed using an antisense Epstein-Barr virus encoded RNA probe. Immunohistochemistry was performed to evaluate the expression of latent membrane protein-1 (LMP-1) and EBV nuclear antigen 2 (EBNA-2). PCR investigation confirmed the presence of EBV in one case. Positive signals confined to tumour cells were present on ISH. None of the tumours showed expression of LMP-1 and EBNA-2. To our knowledge, this is the first report on the presence of EBV in primary adenocarcinoma of the lung in a Caucasian patient. The present study indicates that EBV may infect some lung cancers with a specific tumour pattern.

EBV-expressing AGS gastric carcinoma cell sublines present increased motility and invasiveness.

Tumor invasion marks a critical point in cancer progression; it is a harbinger of morbidity and mortality. Thus, the cellular events that enable the invasive phenotype are under intense investigation. Epstein-Barr virus (EBV) is associated with a number of cancers, including Burkitt lymphoma (BL) and nasopharyngeal carcinoma (NPC) and is suspected to contribute to their tumorigenesis. On average, 8% of gastric carcinomas have been shown to carry this virus. To explore whether the presence of EBV in gastric carcinoma contributes to tumor progression in this predominantly invasive carcinoma, we examined a panel of 2 in vitro EBV-infected human gastric cancer cell line sublines and their mock-infected AGS parental control line. We found EBV infection caused a marked increase in transmigration of a Matrigel barrier (415% and 303%, p < 0.05, for the 2 infected lines). This correlated with increased motility of these sublines (233% and 140%, p < 0.05). As this pattern of increased motility leading to a more pronounced enhancement of invasion has been noted in other tumor cells, we explored the roles of autocrine signaling pathways previously implicated in carcinoma motility and invasion. Inhibitors to the epidermal growth factor receptor (EGFR) (PD153035), phospholipase C (PLC) (U73122), extracellular-signal regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) (PD089035) and PI-3 kinase (Wortmannin) were not informative. These data suggest that EBV increases migration of AGS cells by a mechanism independent of these autocrine growth factor-induced pathways. Instead, we found that the EBV-infected cells presented increased focal adhesion kinase (FAK) phosphorylation. These findings suggest a role for integrin-mediated signaling in promoting EBV-associated invasiveness.

Epstein-Barr virus in gastric carcinoma in Suzhou, China and Osaka, Japan: association with clinico-pathologic factors and HLA-subtype.

Information on geographic differences of Epstein-Barr virus (EBV) positivity and association with HLA in gastric carcinoma are limited. Therefore, the association of gastric carcinoma with EBV was examined in 118 patients from Suzhou, China, where the incidence of nasopharyngeal carcinoma (NPC) is high, and in 216 patients from Osaka, Japan, where the incidence of NPC is low, NPC being one of the EBV-associated carcinomas. Expression of HLA-A2 was also examined in some of these cases. The EBV genome was evidenced by PCR and in the tumor cells by in situ hybridization in 7/90 and 9/151 of cases from Suzhou and Osaka, respectively. Immunohistochemistry revealed that cancer cells in all cases with EBV did not express latent membrane protein-I. Type A was found in all cases positive for EBV. Among several histologic and clinical factors, only age of patients showed a correlation with EBV positivity: patients over 60 showed a higher frequency than patients below 60 years of age (p < 0.05). Typing of the HLA-A locus was possible in 16 cases positive for the EBV genome: 3 of 4 cases from Suzhou and 4 of 7 from Osaka were positive for HLA-A2 products. Severe lymphoid cell infiltration was found in 2 of 7 cases and 1 of 4 cases with and without the HLA-A2 type, respectively. The reported frequency of EBV positivity in Chinese living in Taiwan and in Japanese living in Hawaii is higher than in Suzhou, China, and Osaka, Japan, respectively. Our findings suggest that EBV association with gastric carcinoma is influenced by environmental and cultural factors.