Epithelial-to-mesenchymal transition in human esophageal cancer associates with tumor progression and patient’s survival

The epithelial-mesenchymal transition (EMT) is an essential step in invasion and metastasis of human cancers. Identification of EMT status would help us to properly understand the mechanism of cancer metastasis and progression. In the present study, tissue microarray and immunohistochemical staining of two important markers, E-cadherin and Vimentin, were used to characterize the EMT status in human esophageal cancer. We selected the appropriate cut-off values of expression levels of E-cadherin and Vimentin, and found 63 out of 105 cases of esophageal cancers underwent EMT. And we also found that in the subgroup with (T₃ + T₄), the ratio of patients undergoing EMT was significantly higher than that in the subgroup with (T₁ + T₂) (P = 0.0097), and in the subgroup with metastasis, the ratio of patients undergoing EMT was significantly higher than that in the subgroup with no metastasis (P = 0.0253). The log-rank survival analysis showed that the overall survival rate of the patients undergoing EMT was significantly poorer than that of the patients with wide type status (P = 0.0278, HR = 2.470, 95% CI: 1.971~2.970). In the COX model analysis, we also found that the EMT status of the esophageal cancer patients could be used as an independent risk factor for the prediction of prognosis of this malignancy (P = 0.026, HR = 2.306, 95% CI: 1.103~4.824). Thus, our present study successfully established a method by using tissue microarray and the markers, E-cadherin and Vimentin, to conveniently and properly identify the EMT status in human esophageal cancer, and revealed that the EMT status significantly associated with invasion, metastasis and prognosis in this malignancy.     

B7-H6 expression correlates with cancer progression and patient’s survival in human ovarian cancer

B7-H6, a newly identified B7 family member molecule, binds to its receptor on NK cells, NKp30, and then triggers the anti-tumor NK cell cytotoxicity and leads to the cytokine secretion. As of now, numerous studies have demonstrated that the higher B7-H6 expression could be found in certain human cancers and have important clinical significance. In our present study, we carried out the tissue microarray and the immunohistochemistry assay to investigate the clinical significance of B7-H6 expression in human ovarian cancer. Our results showed that the positive B7-H6 staining was predominantly observed on the membrane and in the cytoplasm of the ovarian cancer cells. In order to further investigate the correlation between clinical parameters and the B7-H6 protein levels in the ovarian tissues, we categorized all the 110 patients into two major subgroups according to the intensity of B7-H6 immunohistochemical staining, i.e., the lower B7-H6 expression group, 34 cases (0 ≤ H-score < 100), and the higher B7-H6 expression group, 76 cases (H-score ≥ 100), and we found that B7-H6 expression in the ovarian cancer tissues is significantly correlated with distant metastasis status (P = 0.028) and FIGO stage (P = 0.031), whereas it is not correlated with patient’s age, tumor size, tumor location, pathological stage or nodal metastasis. The survival analysis demonstrated that the overall survival rate of the subgroup with lower B7-H6 expression is significantly better than that of the subgroup with higher B7-H6 expression (P = 0.0456, Hazard Ratio: 1.707, 95% CI, 1.010-2.885). Thus, our present data revealed that higher B7-H6 expression in ovarian cancer tissues was positively correlated with tumor metastasis and cancer progression, and supports the notion that B7-H6 expression is involved in the progression of human ovarian cancer, the detailed mechanism merits further investigation.     

Hodgkin’s lymphoma in a patient with Jo-1 syndrome

Jo-1 syndrome is an autoimmune disease with autoantibodies against the histidyl tRNA synthetase. Characteristic clinical findings include inflammatory myopathy and interstitial lung disease. We present the first case of a patient with Jo-1 syndrome (positive Jo-1 autoantibodies, myositis, interstitial alveolitis) who developed Hodgkin’s lymphoma of nodular-sclerosing type. Thus, patients with Jo-1 syndrome and immunosuppressive therapy similar to other patients with autoimmune disease are at risk to develop lymphomas and should therefore be monitored carefully.     

Is human kallikrein-11 in gastric cancer treated with surgery and adjuvant chemoradiotherapy associated with survival?

Human kallikreins (hKs) have been reported to be involved in human cancers, and several hKs are promising biomarkers of various cancers, such as prostate, ovarian, breast, and testicular cancer. In the present study, we aimed to evaluate the prognostic value of immunohistochemical expression of hK11 in patients with gastric cancer. The study included 55 (36 men and 19 women; 58 ± 10 years of mean age) patients with gastric cancer treated with surgery and adjuvant chemoradiotherapy. Tissue sections were evaluated immunohistochemically with a monoclonal anti-hK11 antibody. Of the 55 patients, 35 (63.6%) were hK11-positive and 20 (36.4%) were hK11-negative. Disease-free and overall survival rates were significantly higher in patients with hK11 positive than in those with hK-11 negative expression (24 months vs. 11 months, p: 0.043; 29 months vs. 13 months, p: 0.038, respectively). In conclusion, hK11 expression in gastric cancer appears to be associated with a better prognosis. hK11 may be a prognostic biomarker of gastric cancer. On the other hand, it is needed to elucidate the mechanisms underlying the regulation of hK11 expression in gastric cancer.     

Increased expression of 14-3-3β promotes tumor progression and predicts extrahepatic metastasis and worse survival in hepatocellular carcinoma

14-3-3β is implicated in cell survival, proliferation, migration, and tumor growth; however, its clinical relevance in tumor progression and metastasis have never been elucidated. To evaluate the clinical significance of 14-3-3β, we analyzed the association of 14-3-3β expression and clinicopathologic characteristics in primary and subsequent metastatic tumors of hepatocellular carcinoma patients. 14-3-3β was expressed abundantly in 40 of 55 (70.7%) primary tumors. Increased 14-3-3β expression in primary tumors predicted a higher 5-year cumulative incidence of subsequent extrahepatic metastasis, and multivariate analysis revealed 14-3-3β overexpression was an independent risk factor for extrahepatic metastasis. Patients with increased 14-3-3β expression in primary tumors had worse 5-year overall survival rates, and 14-3-3β overexpression was an independent prognostic factor on Cox regression analysis. Furthermore, stably overexpressed 14-3-3β enhanced hepatocellular carcinoma cell migration and proliferation and increased anchorage-independent cell growth. In addition, in vivo study in a nude-mice model showed tumor formation significantly increased with 14-3-3β overexpression. In conclusion, this is the first report to show that increased 14-3-3β expression is associated with subsequent extrahepatic metastasis and worse survival rates, as well as cancer progression of hepatocellular carcinoma. Thus, 14-3-3β may be a novel prognostic biomarker and therapeutic target in hepatocellular carcinoma.     

Cytoplasmic HSP90α expression is associated with perineural invasion in pancreatic cancer

Pancreatic cancer (PC) is an aggressive and devastating disease with a dismal prognosis. The study aimed to investigate the role of HSP90α and PDIA3 in patients with PC. Immunohistochemistry was performed on tissue microarrays containing 186 pairs of PC and normal pancreatic tissues to assess the expression levels of HSP90α and PDIA3. The expression levels of cytoplasmic HSP90α (P = 0.032) and PDIA3 (P = 0.043) in PCs were significantly higher than those in normal pancreas tissues, but nuclear HSP90α showed lower expression in PC tissues (P = 0.002). In addition, cytoplasmic expression of HSP90α and PDIA3 was significantly associated with perineural invasion (PNI) (P = 0.004) and sex (P = 0.014), respectively. These results indicate that cytoplasmic HSP90α may serve as a biomarker for PNI in PCs.     

Altered expression of CD44 and DKK1 in the progression of Barrett’s esophagus to esophageal adenocarcinoma

Barrett’s esophagus (BE) is an acquired condition in which the normal lining of the esophagus is replaced by intestinal metaplastic epithelium. BE can evolve to esophageal adenocarcinoma (EAC) through low-grade dysplasia (LGD) and high-grade dysplasia (HGD). The only generally accepted marker for increased risk of EAC is the presence of HGD, diagnosed on endoscopic biopsies. More specific markers for the prediction of EAC risk are needed. A tissue microarray was constructed comprising tissue samples from BE, LGD, HGD, and EAC. Marker expression was studied by immunohistochemistry using antibodies against CD44, DKK1, CDX2, COX2, SOX9, OCT1, E-cadherin, and β-catenin. Immunostaining was evaluated semi-quantitatively. CD44 expression decreased in HGD and EAC relative to BE and LGD. DKK1 expression increased in HGD and EAC relative to BE and LDG. CDX2 expression increased in HGD but decreased in EAC. COX2 expression decreased in EAC, and SOX9 expression increased only in the upper crypt epithelial cells in HGD. E-cadherin expression decreased in EAC. Nuclear β-catenin was not significantly different between BE, LGD, and HGD. Loss of CD44 and gain of DKK1 expression characterizes progression from BE and LGD to HGD and EAC, and their altered expression might indicate an increased risk for developing an EAC. This observation warrants inclusion of these immunohistochemically detectable markers in a study with a long patient follow-up.     

B4GALNT3 expression predicts a favorable prognosis and suppresses cell migration and invasion via β₁ integrin signaling in neuroblastoma

β1,4-N-acetylgalactosaminyltransferase III (B4GALNT3) promotes the formation of GalNAcβ1,4GlcNAc (LacdiNAc or LDN). Drosophila β1,4-N-acetylgalactosaminyltransferase A (B4GALNTA) contributes to the synthesis of LDN, which helps regulate neuronal development. In this study, we investigated the expression and role of B4GALNT3 in human neuroblastoma (NB). We used IHC analysis to examine 87 NB tumors, and we identified correlations between B4GALNT3 expression and clinicopathologic factors, including patient survival. Effects of recombinant B4GALNT3 on cell behavior and signaling were studied in SK-N-SH and SH-SY5Y NB cells. Increased expression of B4GALNT3 in NB tumors correlated with a favorable histologic profile (P < 0.001, χ² test) and early clinical staging (P = 0.041, χ² test) and was a favorable prognostic factor for survival as evaluated by univariate and multivariate analyses. Reexpression of B4GALNT3 in SK-N-SH and SH-SY5Y cells suppressed cell proliferation, colony formation, migration, and invasion. Moreover, B4GALNT3 increased the LacdiNAc modification of β₁ integrin, leading to decreased phosphorylation of focal adhesion kinase (FAK), Src, paxillin, Akt, and ERK1/2. B4GALNT3-mediated suppression of cell migration and invasion were substantially reversed by concomitant expression of constitutively active Akt or MEK. We conclude that B4GALNT3 predicts a favorable prognosis for NB and suppresses the malignant phenotype via decreasing β₁ integrin signaling.Neuroblastoma (NB) is the most common extracranial solid tumor in childhood, accounting for 8% to 10% of all pediatric malignancies.¹ This tumor arises from primitive neuroepithelial cells of the neural crest.² The behavior of NB is markedly heterogeneous, ranging from highly undifferentiated tumors with very poor outcomes to well-differentiated benign ganglioneuroma or NB that may spontaneously regress with favorable prognosis.³ Half of all patients with newly diagnosed NB are in a high-risk subset with poor overall survival despite intensive therapy. Therefore, it is important to develop useful prognostic tools and to understand NB pathogenesis to help design improved NB therapies.Glycosylation is regulated spatiotemporally during development of the nervous system.⁴ Altered carbohydrate structures on tumors are often associated with tumor metastasis and progression. Tumor-associated carbohydrate epitopes commonly found in cancers include GM2, GD2, GD3, T, Tn, Globo H, Lewis y, sialyl Lewis x, sialyl Lewis a, and polysialic acid.^5,6 Among these, GM2, GD2, and GD3 are expressed in NB.⁶ Changes in expression levels of glycogenes may play an important role in alterations of carbohydrate structures in tumors. However, the differential expression of these molecules in NB and their effects on tumor cell behavior are poorly understood.β1,4-N-acetylgalactosaminyltransferase III (B4GALNT3) has been cloned and is expressed in various tissues.⁷ This enzyme can transfer GalNAc to any nonreducing terminal GlcNAc-β in vitro, resulting in synthesis of GalNAcβ1,4GlcNAc (LacdiNAc or LDN). The terminal β1,4–linked GalNAc of LDN can be recognized by a lectin, Wisteria floribunda agglutinin (WFA). This special terminal β1,4GalNAc structure is found in certain glycoproteins and glycohormones, including lutropin,⁸ thyrotropin,^8–10 proopiomelanocortin,^11–13 and the sorting protein–related receptor SorLA/LR11.¹⁴ SorLA/LR11, highly expressed by neurons in the central and peripheral nervous systems, bears N-linked oligosaccharides modified with terminal β1,4–linked GalNAc-4-SO₄ that can be synthesized by B4GALNT3 in CHO cells.¹⁴ In Drosophila, β1,4-N-acetylgalactosaminyltransferase A (B4GALNTA) catalyzes synthesis of LacdiNAc on glycoconjugates.¹⁵ Drosophila mutants deficient in B4GALNTA have defects in behavior and in the neuromuscular system.¹⁶ In the present study, we hypothesized that B4GALNT3 may be involved in the pathogenesis of NB development.     

PinX1 is up-regulated and associated with poor patients’ survival in gliomas

PinX1, a conserved nuclear protein, could maintain telomere integrity and plays an important role in regulating telomerase activity. It has been reported that the expression of PinX1 is down-regulated in some cancer and associated with cancer prognosis. However, the value of PinX1 in gliomas has not been studied. In this study, two independent retrospective gliomas cohorts with the corresponding gliomas tissue microarrays (TMAs) were established to detect the expression level of PinX1 and the correlation of PinX1 expression with the clinicopathological features and the patients’ survival. Compared with non-cancerous brain tissues, PinX1 protein levels were remarkably up-regulated in gliomas (P = 0.001), and further increased from benign gliomas tissues to malignant gliomas tissues (P = 0.090). Moreover, high PinX1 expression was significantly positively associated with gliomas WHO grade in the training set (P = 0.019) and the validation set (P = 0.037). High PinX1 expression significantly correlated with a worse 5-year overall (P = 0.016) and disease-specific survival (P = 0.026). Simultaneously, the multivariate COX regression analysis showed that PinX1 was an independent unfavorable prognostic factor for 5-year overall survival (hazard ratio (HR) = 2.078, P = 0.015) and disease-specific survival (HR = 2.429, P = 0.012) after adjusting with age, sex and WHO grade in gliomas. In conclusion, PinX1 expression may serve as a prognostic and predictive biomarker for gliomas.     

B7-H4’s Role “Beyond the Tumor”

B7-H4 is a ligand in the B7 costimulatory family, executing suppressive function on the immune system in many diseases, such as cancer, allograft rejection, and autoimmune diseases. The receptor for this molecule has yet to be clarified. The engagement of B7-H4 inhibits proliferation of immune cells by stopping the cell cycle at the G0/G1 phase and leads to apoptosis via the Fas/FasL pathway consequently accelerating tumor progression and alleviating allograft rejection. The pathogenic role of B7-H4 in tumors has been widely established, but few studies have focused on its function in other disorders. Here, we review recent advances in our understanding of B7-H4 biology in disease settings other than tumors and document the beneficial values to treat those diseases by targeting this molecule and related signaling pathways.     

Gene expression in 16q is associated with survival and differs between Sørlie breast cancer subtypes

We have investigated the relationship between gene expression and chromosomal positions in 402 breast cancer patients. Using an overrepresentation approach based on Fisher's exact test, we identified disproportionate contributions of specific chromosomal positions to genes associated with survival. Our major finding is that the gene expression in the long arm of chromosome 16 stands out in its relationship to survival. This arm contributes 36 (18%) and 55 (11%) genes to lists negatively associated with recurrence-free survival (set to sizes 200 and 500). This is a highly disproportionate contribution from the 313 (2%) genes in this arm represented on the used Affymetrix U133A and B microarray platforms (Bonferroni corrected Fisher test: P < 2.2 x 10(-16)). We also demonstrate differential expression in 16q across tumor subtypes, which suggests that the ERBB2, basal, and luminal B tumors progress along a high grade-poor prognosis path, while luminal A and normal-like tumors progress along a low grade-good prognosis path, in accordance with a previously proposed model of tumor progression. We conclude that important biological information can be extracted from gene expression data in breast cancer by studying non-random connections between chromosomal positions and gene expression. This article contains Supplementary Material available at http://www.interscience.wiley.com/jpages/1045-2257/suppmat.     

Ticagrelor inhibits platelet–tumor cell interactions and metastasis in human and murine breast cancer

Activated platelets promote the proliferation and metastatic potential of cancer cells. Platelet activation is largely mediated through ADP engagement of purinergic P2Y12 receptors on platelets. We examined the potential of the reversible P2Y12 inhibitor ticagrelor, an agent used clinically to prevent cardiovascular and cerebrovascular events, to reduce tumor growth and metastasis. In vitro, MCF-7, MDA-MB-468, and MDA-MB-231 human mammary carcinoma cells exhibited decreased interaction with platelets treated with ticagrelor compared to untreated platelets. Prevention of tumor cell–platelet interactions through pretreatment of platelets with ticagrelor did not improve natural killer cell-mediated tumor cell killing of K562 myelogenous leukemia target cells. Additionally, ticagrelor had no effect on proliferation of 4T1 mouse mammary carcinoma cells co-cultured with platelets, or on primary 4T1 tumor growth. In an orthotopic 4T1 breast cancer model, ticagrelor (10 mg/kg), but not clopidogrel (10 mg/kg) or saline, resulted in reduced metastasis and improved survival. Ticagrelor treatment was associated with a marked reduction in tumor cell–platelet aggregates in the lungs at 10, 30 and 60 min post-intravenous inoculation. These findings suggest a role for P2Y12-mediated platelet activation in promoting metastasis, and provide support for the use of ticagrelor in the prevention of breast cancer spread.     

The interplay between partners’ responsiveness and patients’ need for emotional expression in couples coping with cancer

The central aim of this longitudinal observational study was to test whether patients with a high need for emotional expression are especially sensitive to their partners’ responsive behavior, and therefore at risk for depressive symptoms when responsiveness is withheld. Patients with colorectal cancer and their partners (n = 58) participated in a longitudinal study (3, 5 and 9 months after the diagnosis). Additionally to self-report measurements (i.e., patients’ need for emotional expression, patients’ depressive symptoms and patients’ relationship satisfaction) couples were videotaped discussing cancer-related concerns. External observers coded partners’ responsiveness (i.e., understanding, validation and caring) and patients’ self-disclosures. Partner responsiveness predicted lower levels of depressive symptoms over time in patients who had a relatively high need for emotional expression above and beyond the effect of relationship satisfaction. We demonstrated that partners’ understanding and validation are more important in explaining patients’ depressive symptoms than partners’ caring behavior. Our findings highlight the importance of the relational context in improving adaptation to cancer taking into account individual differences.     

P15, MDM2, NF-κB,and Bcl-2 expression in primary bone tumor and correlation with tumor formation and metastasis

Primary bone tumor is one of the most common malignant tumors in skeletal system. It seriously affected bone movement and development with unclear pathogenesis. In this paper, rabbit VX-2 malignant bone tumor model was applied to explore apoptotic genes P15, MDM2, NF-κB and Bcl-2 correlation with primary bone tumor occurrence and metastasis. 0.3 ml rabbit VX-2 tumor cell suspension (1×10⁶/ml) was injected to the marrow cavity of the right tibia condyle to establish the rabbit malignant bone tumor model, while equal amount of the saline was injected to the left tibia as control. Real-time PCR was applied to determine P15, MDM2, NF-κB and Bcl-2 expression level. Immunohistochemistry was performed to detect the abovementioned genes expression in lung, stomach, kidney and bladder. Compared with control, P15 expression level in the inoculation site surrounding tissues decreased obviously following the inoculate time elongation (P<0.05), while Bcl-2, MDM2 and NF-κB expression significantly increased (P<0.05). Bcl-2 showed significant correlation with MDM2 and NF-κB (P<0.05). At the 2, 4, 6 weeks, Bcl-2, MDM2 and NF-κB in lung, Bcl-2 in kidney, and Bcl-2 and MDM2 in bladder positively expressed (P<0.05), whereas P15 gene exhibited no significant positive expression in these tissues (P>0.05). P15, MDM2, NF-κB, and Bcl-2 genes expression levels can effectively reflect malignant bone tumor growth of rabbit tibia. MDM2, NF-κB and Bcl-2 genes involved in primary bone tumors metastasis directly. It has important clinical significance for early diagnosis and treatment of primary bone tumor.     

Immunohistochemical study of expression of trophoblasticβ 1-globulin in gastric epithelium of a patient with stomach cancer

P. A. Gertsen Moscow Oncologic Research Institute. N. V. Sklifosovskii Emergency Aid Research Institute, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. K. Permyakov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 9, pp. 330–332, September, 1991.