Expression analysis of BRUCE protein in esophageal squamous cell carcinoma

Apoptosis is a form of cell death in response to diverse stressful physiological or pathological stimuli. One of the most important gene families involved in apoptosis is inhibitors of apoptosis. As a member of inhibitors of apoptosis, BRUCE can suppress apoptosis and promote cell division. Because esophageal squamous cell carcinoma (ESCC) cells, as well as other cancer cells, are immortal, our aim in this study was to analyze BRUCE protein expression in ESCC and evaluate its correlation with tumoral clinicopathologic features. Fifty ESCC specimens were examined for BRUCE protein expression using immunohistochemistry. A defined scoring method was applied. BRUCE protein was detected in 82% of tumors. Tumor progression stage and invasion depth correlated significantly with BRUCE protein expression (P = .019 and .005, respectively). Furthermore, association of BRUCE expression with tumor location was near significant (P = .058). The correlation of BRUCE overexpression in ESCC and disease aggressiveness may confirm the importance of BRUCE in ESCC progression and invasiveness. Therefore, BRUCE protein may be a molecular marker for aggressive ESCC and, thus, a potential therapeutic target to inhibit tumor cell progression and invasion.     

子宫颈、食管鳞状细胞癌人乳头状瘤病毒感染的比较研究

目的 探讨同一地区人乳头瘤病毒在宫颈鳞状细胞癌的感染特征及与食管鳞状细胞癌的关系.方法 采用PCR技术和快速导流杂交基因芯片技术分别对来自河南同一地区的75例宫颈鳞癌患者和78例食管鳞癌患者进行HPV 分型检测.结果 来自同一地区的宫颈、食管鳞癌组织中,HPV16型阳性检出率均最高,分别为83%(58/70)和82%(49/60),其余阳性类型宫颈鳞癌中依次为HPV58、18、31、6、52、33、11,分别占14%(10/70),9%(6/70),7%(5/70),6%(4/70),6%(4/70),4%(3/70),4%(3/70);食管鳞癌中依次为HPV18、58、31、6、11,分别占15%(9/60),8%(5/60),7%(4/60),5%(3/60),5%(3/60).对HPV16阳性的宫颈、食管鳞癌标本DNA进行HPV16致癌基因E6/E7扩增进行验证,阳性率为100%.结论 同一地区居民宫颈(食管)鳞癌HPV感染亚型相似,提示HPV可能是这两种肿瘤的共同致病因素,HPV16可能在宫颈和食管癌变过程中起重要作用,也为其生物防治提供了重要理论依据.     

Pathways of vulvar intraepithelial neoplasia and squamous cell carcinoma

Vulvar squamous cell carcinoma (VSCC) accounts for >90% of the malignant tumours of the vulva. Most VSCCs originate in intraepithelial lesions, named vulvar intraepithelial neoplasia (VIN), that precede the development of VSCC by a variable period of time. Strong evidence has accumulated showing that there are two different aetiopathogenic pathways for the development of VSCC and VIN, one associated with infection by human papillomavirus (HPV), and a second independent of HPV infection. These two different types of VSCC have different epidemiological, pathological and clinical characteristics, and should therefore be considered as two separate entities. Histologically, HPV‐associated VSCCs are of the basaloid or warty type, and arise from VIN of the usual type. Inactivation of p53 and the retinoblastoma tumour suppressor gene product by the viral gene products E6 and E7 is involved in the process of malignant transformation. HPV‐independent VSCCs are histologically keratinizing, are associated with differentiated VIN and lichen sclerosus, and frequently show mutations of p53. p16^INK4a and p53 immunostaining can be useful for classifying VSCC into HPV‐associated or HPV‐independent. Although large, multicentre studies are needed to definitively assess the involvement of HPV in the prognosis of VSCC, most studies have not found clear differences in survival between HPV‐associated and HPV‐independent tumours.     

Gastric small-cell undifferentiated carcinoma with adeno and squamous cell carcinoma components

A unique gastric tumor is reported. A large portion of the tumor consisted of a diffuse sheet of undifferentiated cells reminiscent of a small cell undifferentiated carcinoma. The tumor cells showed a few dense core granules and a poorly developed attachment apparatus by electron microscopy. In addition, small portions of the tumor showed adenocarcinoma and squamous cell carcinoma. Another noteworthy finding was that some of the metastatic hepatic nodules consisted of relatively monotonous polygonal cells with a distinct cord-like pattern showing argentaffinity and argyrophilia which were indicative of an atypical carcinoid. This case is a rare example of a gastric tumor with differentiation towards endocrine as well as adeno and squamous cell carcinoma.     

Physical status of HPV-16 in esophageal squamous cell carcinoma.

BACKGROUND: Infection with high-risk human papillomavirus (HPV) has been implicated as one of the risk factors of esophageal squamous cell carcinoma (ESCC). Integration of viral DNA into host genome is essential for carcinogenesis since it promotes disruption of the HPV E2 gene, leading to abnormal expression of E6 and E7 oncoproteins. OBJECTIVES AND STUDY DESIGN: To investigate the viral integration status of HPV-16 infection in ESCC, 35 HPV-positive ESCC specimens collected from Chinese patients were subject to real-time quantitative PCR for determination of physical status of HPV-16 by analyzing the ratios of E2 to E6 genes. RESULTS: Our results showed that only 8.6% (3/35) of the HPV-16 positive specimens harbored exclusively the episomal form (i.e. E2/E6 ratio > or = 1), whereas the remaining 91.4% contained either only the integrated form (5.7%, with E2/E6 ratio = 0) or a mixture of episomal and integrated forms of viral molecules (85.7%, with E2/E6 ratios > 0 but < 1). Amongst the 30 cancer specimens carrying mixed integrated and episomal forms, 28 had E2/integrated E6 ratios of less than 1, indicating a predominance of integrated form of viral genes in these lesions. CONCLUSION: Our finding of frequent integration of viral DNA in the host genome suggests that integration HPV-16 is common in ESCC from Chinese patients and implies that HPV infection may play a role in the pathogenesis of ESCC.     

Candidate susceptibility variants for esophageal squamous cell carcinoma

Esophageal cancer is common worldwide, and often fatal. The major histological subtype is esophageal squamous cell carcinoma (ESCC). ESCC shows familial aggregation and high heritability. Mutations in RHBDF2 cause tylosis, a very rare disorder characterized by high life‐time risk of ESCC, but no other well‐established predisposition genes have been identified. To identify candidate susceptibility variants for ESCC we utilized the Population Information System and the Finnish cancer registry to find study materials by clustering ESCC patients by family name at birth and municipality at birth. We collected archival tissue material and exome sequenced a total of 30 ESCC cases. We prioritized shared, deleterious and rare variants that were significantly enriched in our sample set compared to Finnish and population subset specific controls. Six variants passed filtering, the most frequent being a nonsense mutation in DNAH9 (p.Tyr1573Ter) found in four unrelated patients. DNAH9 has been reported to be frequently lost in ESCC tumors. In this study, one patient's tumor showed loss of the wild type allele of DNAH9 suggesting a tumor suppressive function. A missense variant in GKAP1 was shared by three patients, and missense variants in BAG1, NFX1, FUK, and DDOST by two each. EP300 which has previously been implicated in the genesis of ESCC had a missense variant segregating in three affected individuals in a single family. If validated in independent patient sets, these variants could serve as a tool towards prevention and early diagnosis of ESCC.     

Small cell (endocrine cell) carcinoma of the gallbladder with squamous and adenocarcinomatous components.

Small cell (endocrine cell) carcinoma of the gallbladder in a 62-year-old woman is reported. The palliative cholecystectomy specimen revealed a submucosally invading tumor with extensive hemorrhagic necrosis. At autopsy, performed five months after surgery, a huge tumor measuring 14 x 12 x 8 cm was located at the liver hilus. No signs or symptoms related to overproduction of hormones were recorded throughout her illness. Neither lung lesions nor gall stones were identified. Histologically, diffuse proliferation of small, spindle-shaped atypical tumor cells with numerous mitoses was evident. Intraepithelial tumor cell proliferation in the gallbladder mucosa was seen focally. The neuroendocrine nature of the tumor cells was confirmed by the histologic pattern of growth with pseudo-rosette formation, positive reaction for Grimelius' argyrophilia, neuron-specific enolase and Leu 7, and ultrastructural demonstration of neuroendocrine-type granules. Immunostaining for a variety of hormones was all negative. Characteristically, foci with squamous and adenocarcinomatous differentiation were identified in the tumor tissue. The glandular components were immunoreactive for carcinoembryonic antigen, secretory component, epithelial membrane antigen and CA19-9. The histogenesis and totipotentiality of the neoplasm were discussed.     

Cytogenetic heterogeneity and progression of esophageal squamous cell carcinoma

It is widely believed that most human tumors, including esophageal squamous cell carcinoma (ESCC), arise through multistep genetic and cytogenetic alterations. The time sequence of these alterations, however, is still unknown. The present study was designed to differentiate common early changes from uncommon later ones with combined comparative genomic hybridization (CGH) and ploidy analyses in multiple or single samples of 12 ESCCs. We first demonstrated that the mean copy numbers of chromosomes 3 and 11, determined directly by fluorescence in situ hybridization, showed linear correlation with the mean copy numbers calculated from the G/R ratio of CGH and DNA ploidy (R(2)=0.714, P<0.0001). On this basis, we estimated the absolute copy numbers of chromosomal parts by applying the ploidy-dependent threshold criteria to the G/R ratio data after the criteria were corrected by the percentage of tumor cells in each sample. One-copy changes in the DNA-diploid stage may give large shifts of the G/R ratio, even after tetraploidization, whereas those after tetraploidization undergo small shift. Using the tumors with multiple samples, it was actually demonstrated that most of the gains common to the samples in individual tumors showed the large shifts. Though early changes varied from tumor to tumor in the nine informative cases, it was found that gains of 3q (5/7: number of cases with large-shift 3q+/total number of cases with 3q+), 8q (3/4), 11q13 (4/5), and 14q (3/4) were early events, while losses of 3p (2/8), 5q (1/5), 13q (1/5), and 21q (1/5), and gains of 1p (1/4) and Xq (1/4) were later events in progression of individual tumors.     

Accumulation of p53 in esophageal squamous cell carcinoma

D-mannoheptulose is a specific inhibitor of D-glucose phosphorylation by hexokinase isoenzymes. In the present study, the phosphorylation of this heptose was investigated by either a spectrophotometric or radioisotopic procedure. Using yeast hexokinase, the phosphorylation of 25 mM D-mannoheptulose only represented 0.02% of that of 5 mM D-glucose. Such a percentage was increased to 3.93% in the case of bovine heart hexokinase. In the latter case, the Km for D-mannoheptulose was close to 0.2 mM and both D-glucose (0.1-1.0 mM) and D-glucose 6-phosphate (also 0.1-1.0 mM) inhibited the phosphorylation of the heptose (0.03-0.60 mM). Human B-cell glucokinase also catalyzed the phosphorylation of D-mannoheptulose (0.1 mM), which was now increased in a bell-shaped manner by D-glucose (1.0-20 mM). Likewise, rat parotid gland, liver and pancreatic islet homogenates catalyzed the phosphorylation of D-[3H]mannoheptulose. The results obtained in these three tissues differed from one another by their absolute values (per mg wet wt.), relative values (by reference to the phosphorylation rate of 10 mM D-glucose), and sensitivity to inhibition by D-glucose (10 mM).     

Small cell carcinoma of the ureter with squamous cell and transitional cell carcinomatous components associated with ureteral stone.

We report a case of primary small cell carcinoma of the ureter with squamous cell and transitional cell carcinomatous components associated with ureteral stone, which is unique in that the patient has remained free of tumor recurrence for 36 months after the surgery without adjuvant chemotherapy or radiotherapy. A 60-yr-old man presented himself with a right flank pain. Computed tomography revealed an ill-defined mass and a stone in the lower one third of the right ureter, and hydronephroureterosis above the stone-impacted site. The patient underwent right nephroureterectomy and stone removal. Upon gross examination, a 3.8 x 1.8 x 1.2 cm white and partly yellow mass was noted in the anterior part of the ureter, resulting in indentation of the ureteral lumen on the posterior side. Light microscopic examination revealed that the mass was mainly composed of small cell carcinoma, and partly squamous cell and transitional cell carcinomatous components. The overlying ureteral mucosa and renal pelvis also contained multifocal dysplastic transitional epithelium and transitional cell carcinoma in situ. There was no vascular invasion, and the surgical margins were free of tumor. The small cell carcinomatous component was positive for chromogranin, neuron specific enolase, synaptophysin, and pancytokeratin but negative for high molecular-weight cytokeratin (K-903) by immunohistochemistry.     

Overexpression of matriptase correlates with poor prognosis in esophageal squamous cell carcinoma

Matriptase is one of the type II transmembrane serine proteases and is known to be involved in cancer progression. Increased matriptase expression has been reported in a variety of human cancers, and its association with poor prognosis has been highlighted in some cancer types. However, its exact role in cancer progression and its effect on patient survival in esophageal squamous cell carcinoma (ESCC) are still unclear. We performed immunohistochemical staining of matriptase in 171 ESCC samples after antibody validation and evaluated the association of its expression with clinicopathological parameters and prognosis. High matriptase expression was observed in 38.6 % (66/171) of ESCC samples and more frequently in N3 stage and in poorly differentiated tumors. Both overall survival (OS) and disease-free survival (DFS) were significantly lower for patients with high expression of matriptase than for patients with low expression (5-year OS rate, 38.6 vs 55.3 %; p?=?0.034 and 5-year DFS rate, 30.5 vs 49.4 %; p?=?0.007). High matriptase expression was an independent prognostic factor for OS [hazard ratio (HR), 1.65 (95 % confidence interval (CI), 1.01–2.68); p?=?0.045] and for DFS [HR, 1.79 (95 % CI, 1.14–2.81); p?=?0.012]. In conclusion, higher expression of matriptase is an independent prognostic factor involved in the progression of ESCC, which suggests that matriptase is a factor in ESCC tumor progression and also a potential molecular therapeutic target.     

Expression of HLA-G is associated with prognosis in esophageal squamous cell carcinoma

The detection of HLA-G expression might serve as a clinical marker in the diagnosis or prediction of clinical outcomes for certain types of carcinoma. The aim of this study was to determine whether the detection of HLA-G has any important clinical applications for patients with esophageal squamous cell carcinoma (ESCC) by using immunohistochemical methods. We observed that the HLA-G protein was expressed in 90.9% (110/121) of the primary sites of ESCC but not in the normal esophageal tissues. The expression of HLA-G in the tumors was significantly correlated with histologic grade, depth of invasion, nodal status, host immune response, and clinical stage of disease. Patients with positive HLA-G expression had a significantly worse prognosis. In multivariate analysis, HLA-G was an independent prognostic factor. Our results indicate that expression of HLA-G is a characteristic feature of ESCC and suggest that immunostaining by anti-HLA-G antibodies may be a potentially useful prognostic indicator.     

FHIT expression and hypermethylation in esophageal squamous cell carcinoma

The expression of the fragile histidine triad (FHIT) gene has been proposed to play an important role in early events of carcinogenesis and to be correlated with the progression or clinical outcomes of various cancers. Attention has focused recently on the regulation of FHIT expression, and loss of heterozygosity or hypermethylation of the CpG island in the promoter region has been suggested as clues to a possible mechanism. Methylation status and FHIT expression were investigated in the present study to clarify the clinicopathologic impact of FHIT in vivo. One hundred and five patients with esophageal cancer were admitted to the study. Cancer tissues were immunohistochemically stained for FHIT, and FHIT methylation status was examined in 36 patients by the methylation-specific polymerase chain reaction. FHIT methylation and expression were analyzed with respect to both clinicopathologic parameters and their interactions. Tissue specimens from 35 of the 105 patients (33.3%) stained positively for FHIT. In contrast, the CpG island in the FHIT promoter region was hypermethylated in 25 of the 36 (69.4%) analyzed cases of esophageal cancer. Hypermethylation was significantly correlated with the deletion of FHIT protein expression (P<0.001). FHIT hypermethylation was not associated with any clinicopathologic parameters. In contrast, deletion of FHIT expression significantly promoted tumor invasion (P<0.05) and lymphatic vessel invasion (P<0.01). Lymph node metastasis also appeared higher in the absence of FHIT protein expression, but the result was not significant (P=0.069). Patients with a preserved FHIT gene expression possibly exhibited an improved prognosis compared with those with deleted FHIT expression (P=0.093). Hypermethylation of the FHIT promoter region may be a mechanism for regulating FHIT expression. FHIT gene expression was closely correlated with cancer progression, as indicated by tumor invasion and lymphatic spread, and it may provide insight into the mechanism of progression of esophageal cancer.     

Cytogenetic studies of primary cultures of esophageal squamous cell carcinoma

Cytogenetic analysis was performed on primary cultures of 21 squamous cell carcinomas of the esophagus (SCCE). Seven cases exhibited mosaic clonal chromosome abnormalities distributed as follows: two contained tetraploid cell populations, one with t(3;7)(p21;q11); two showed loss of the Y chromosome, one with double minutes; single cases demonstrated der(11)t(4;11)(q?27;q23); add(1)(p35) and del(4)(p12); and del(7)(p13), del(7)(q22q34), and der(11)t(7;11)(p?15;p?13). The remaining 14 cases had apparently normal karyotypes, possibly derived from stromal elements. These results demonstrate numerical abnormalities and the multiple occurrence of rearrangements involving chromosomes 7 and 11 in SCCE.     

High endothelial venules associated with better prognosis in esophageal squamous cell carcinoma

BackgroundHigh endothelial venules (HEVs) are specialized microvessels for recruiting naïve T cells and B cells from the circulation into secondary lymphoid organs. Its involvement in esophageal squamous cell carcinoma (ESCC) is still unknown. This study mainly investigated the possible presence of HEVs in ESCC and explore its relationship with prognosis.MethodFormalin fixed paraffin embedded (FFPE) tissue samples of 52 ESCC patients were stained with immunohistochemically (IHC) to assess the association of HEVs with histological and clinical factors by immunohistochemistry. Furthermore, multiplexed immunofluorescence was performed to explore the microenvironment around HEVs.ResultHEVs was widely present in ESCC and was significantly associated with better overall survival (OS). In addition, multiplexed immunofluorescence imaging demonstrated that HEVs is mainly present in the tertiary lymphoid structures (TLS) of the tumor and is surrounded by a large number of lymphocyte cells.ConclusionHEVs represent a better prognostic factor in ESCC.     

Clinicopathology significance of podoplanin immunoreactivity in esophageal squamous cell carcinoma

Backgroud and aim: Podoplanin (D2-40) is a specific marker for lymphatic endothelium. The vast majority of previous studies on podoplanin immunostaining in esophageal squamous cell carcinoma (ESCC) focused on identifying lymphatic vessel invasion (LVI) and counting lymphatic vessel density (LVD) and had contradictory results. Recent studies show podoplanin expression on cancer cells or tumor stroma in several cancers, which have specific significance; but the status in ESCC remains unclear. Therefore, the aim of this study was to further study and summarize the clinicopathological significance of podoplanin immunoreactivity in ESCC. Materials and methods: We examined podoplanin expression in tissue specimens from 107 patients with ESCC by immunohistochemistry. Podoplanin positive lymphatic vessels in intratumoral and peritumoral tissues and podoplanin positive expression in cancer cells and tumor stroma were analyzed, and correlated with clinicopathologic parameters and three-year overall and free-disease survival. Results: 34 (31.8%) and 28 (26.2%) of 107 specimens had podoplanin positive expression in cancer cells and tumor stroma, respectively. Logistic regression analysis showed high intratumoral lymphatic vessel density (I-LVD) and podoplanin positivity in cancer cells were increased risks of lymph node metastasis (LNM) (OR = 2.45, P = 0.03; OR = 0.35, P = 0.01, respectively). Survival analysis showed that I-LVD was a significant factor related to poor three-year overall and free-disease survival (P = 0.04, P = 0.03, respectively). Conclusions: Previous data and our results show that podoplanin seems to be a useful marker to predict LNM, recurrence, and worse prognosis in ESCC; in particular, LVI, high I-LVD, and podoplanin positivity in cancer cells are associated with LNM, recurrence and overall survival.