Alteration of HLA‐F and HLA I antigen expression in the tumor is associated with survival in patients with esophageal squamous cell carcinoma

Alteration of human leukocyte antigen (HLA) expression, such as decreased HLA I (HLA‐A, ‐B and ‐C) antigens and elevated nonclassical HLA I antigens (HLA‐E, ‐F and ‐G), was reported to have an unfavorable prognosis in various cancers. In our study, HLA‐F expression in 105 primary esophageal squamous cell carcinoma (ESCC) lesions and 62 case‐matched adjacent normal tissues, and HLA I antigens among 68 cases were analyzed by immunohistochemistry. Data revealed that HLA‐F expression was observed in 58.1% (61/105) of the ESCC lesions and in 54.8% (34/62) of the normal esophageal tissues. Among the 62 case‐matched samples, HLA‐F expression (lesion vs. normal tissue) was upregulated, unchanged and downregulated in 13 (21.0%), 6 (9.6%) and 43 (69.4%) cases, respectively. Patients with HLA‐F positive had a worse survival than those with HLA‐F negative (p = 0.040). Patients with upregulated HLA‐F expression (lesion vs. normal tissue) had significantly worse survival than those with HLA‐F unchanged and downregulated (p = 0.010). Furthermore, decreased HLA I expression was observed in 41.2% (28/68) patients and was with worse prognosis in comparison to those with preserved HLA I expression (p = 0.001). Multivariate analysis using Cox's proportional hazards model revealed that upregulated HLA‐F expression (p = 0.026) and downregulated HLA I expression (p = 0.013) could be an independent unfavorable prognostic factor. In conclusion, our study provided the evidence that alteration of HLA I and HLA‐F antigen expression was associated with survival in patients with ESCC.     

Expression of RCAS1 in esophageal squamous cell carcinoma is associated with a poor prognosis

BACKGROUND: Receptor-binding cancer antigen expressed on SiSO cells (RCAS1) has been reported to act as a ligand for a receptor present on normal peripheral lymphocytes and to induce apoptotic cell death. We aimed to elucidate the significance of RCAS1 expression in human esophageal squamous cell carcinomas (ESCC). METHODS: RCAS1 expression was examined immunohistochemically in surgically resected esophageal carcinoma tissues from 114 patients. We also examined the relationships between RCAS1 expressions, the tumor Ki-67 indices (a marker of proliferation), and the number of CD8+ tumor-infiltrating lymphocytes (TILs). RESULTS: RCAS1 immunoreactivity was detected in the membranes and cytoplasm of the tumor cells. Of the 114 esophageal carcinomas, 39 (34.2%) were strongly positive for RCAS1 immunostaining on the membranes of the cancer cells, 41 (36.0%) were weakly positive, and 34 (29.8%) were negative. A comparison of RCAS1 expression and clinicopathological characteristics in all 114 patients revealed significant associations between RCAS1 expression and lymph node status (P < 0.05), and pathologic stage (P < 0.05). The survival rates of patients with RCAS1-negative tumors were significantly higher than those of patients with both RCAS1-weak positive tumors and RCAS1-strong positive ones (log-rank P < 0.05). Multivariate analysis revealed that RCAS1 positivity was an independent prognostic factor (P < 0.05). The relationship between RCAS1 expression and the numbers of CD8+ T-cells in the primary tumors revealed that RCAS1-negative tumors tended to contain more of these cells than both RCAS1-weak positive tumors and RCAS1-strong positive ones (P = 0.2495). CONCLUSIONS: RCAS1 may play a significant role in tumor progression via an immune escape mechanism; thus, RCAS1 expression could be used as a predictor of poor prognosis in patients with ESCC.     

Downregulation of HLA Class I molecules in the tumour is associated with a poor prognosis in patients with oesophageal squamous cell carcinoma

As antigenic peptides in the context of human leukocyte antigen (HLA) class I molecules are recognised by cytotoxic T lymphocytes (CTL), the downregulation of HLA class I molecules is one of the reasons why tumour cells can evade CTL-mediated anti-tumour immunity. In this study, we investigated HLA class I expression in oesophageal squamous cell carcinoma (ESCC) (n=70) and in their metastatic lesions (lymph nodes (n=40) and liver (n=3)), by immunohistochemistry with anti-HLA class I monoclonal antibody (EMR8-5). As a result, the downregulation of HLA class I expression in primary lesions of ESCC was observed in 43%, and that in metastatic lymph nodes was noted in 90%. Furthermore, patients with preserved HLA class I expression in primary tumours showed a better survival in comparison to those with downregulated HLA class I molecules (P<0.01). Furthermore, multivariate analysis using Cox's proportional hazards model revealed that the downregulated expression of HLA class I in primary lesions was an independent, unfavourable prognostic factor (P<0.01). In conclusion, the downregulation of HLA class I expression frequently occurred in primary tumour and, to a greater extent, in metastatic lesions of patients with ESCC and was associated with patient survival.     

EphA2 overexpression correlates with poor prognosis in esophageal squamous cell carcinoma

EphA2 is a member of the Eph family of receptor tyrosine kinases, which interact with cell-bound ligands known as ephrins. EphA2 expression was investigated by immunohistochemistry with an anti-EphA2 monoclonal antibody in 80 patients with esophageal squamous cell carcinoma (ESCC) who had undergone surgery. EphA2 overexpression was positive in 40 of the 80 patients (50%). A significant correlation was observed between EphA2 expression and regional lymph node metastasis (p=0.023), number of lymph node metastases (p=0.011) and poor degree of tumor differentiation (p=0.004). The survival rates of EphA2-positive patients were poorer than those of EphA2-negative patients (p=0.014). The 5-year survival rate of patients without EphA2 overexpression was 68%, whereas that of patients with EphA2 overexpression was 29%. EphA2 expression was also investigated in 7 ESCC cell lines (TE-1, -2, -8, -13, -15, TT and TTn) and 1 immortalized human esophageal keratinocyte cell line (CHEK-1). Western blotting revealed different levels of EphA2 expression in the 8 cell lines. EphA2 was expressed at a high level in the ESCC cell lines compared to CHEK-1. EphA2 phosphorylation was demonstrated in all cell lines. Northern blot analysis showed that EphA2 mRNA expression in TE-1 was greater than that in the other ESCC cell lines. The observation of small gaps on Western blot analysis of the ESCC cell lines suggests that there may be a mechanism for EphA2 regulation at the point of translation. In conclusion, EphA2 overexpression appears to be related to poor degree of tumor differentiation and lymph node metastasis in ESCC. Consequently, patients with EphA2 overexpression have a poorer prognosis than those without. EphA2 is a potential target to prevent ESCC cells spreading into the lymphatic drainage.     

Bcl-X expression in esophageal squamous cell carcinoma: association with tumor progression and prognosis

BACKGROUND AND OBJECTIVES: Bcl-2 family proteins are regulators of programmed cell death and important in the development and progression of human various tumors. The role of these proteins in the development, progression and differentiation of esophageal squamous cell carcinoma (ESCC) is unclear. METHODS: We investigated the expression of Bcl-2, Bcl-X, and Bax using immunohistochemistry in 86 ESCCs, and scored the expression by the weighted score. RESULTS: Bcl-2 expression related to pT category (P=0.043) and histological grade (P = 0.001). Bcl-X expression related to pT category (P = 0.003), pN category (P = 0.041) and the number of positive nodes (P = 0.036), and had a tendency to relate to histological grade (P = 0.086). Bax expression had a tendency to relate to pN category (P = 0.081). The inverse relationship between Bcl-2 and Bcl-X expression was detected (P = 0.001), while the positive one between Bcl-X and Bax expression was detected (P = 0.014). Patients with low Bcl-X weighted score had a significantly longer survival compared with those with high Bcl-X weighted score. Multivariate analysis revealed Bcl-X expression as the independent prognostic factors (P = 0.022). CONCLUSION: These results imply that Bcl-2 family proteins, especially Bcl-X, may contribute to the progression in ESCC.     

Phosphorylated AKT1 is associated with poor prognosis in esophageal squamous cell carcinoma

Background

The epidermal growth factor receptor (EGFR) signaling pathway is important in regulating biological behaviors in many malignancies. We explored whether expression and activation of EGFR and several components on its downstream pathways have prognostic significance in patients with esophageal squamous cell carcinoma (ESCC).

Methods

Expression of EGFR, phosphorylated (p)-EGFR, AKT1, p-AKT1, AKT2, p-AKT2, ERK1, ERK2, p-ERK1/2, STAT3, and p-STAT3 was assessed by immunohistochemical analysis of tissue microarrays for 275 ESCC patients who had undergone complete three-field lymphadenectomy. Spearman rank correlation tests were used to determine the relationships among protein expression, and Cox regression analyses were performed to determine the prognostic factors on overall survival (OS).

Results

p-EGFR expression was correlated statistically with all of the other phosphorylated markers. Gender, N stage, and p-AKT1 expression were found to be independent prognostic factors for OS. Increased expression of p-AKT1 was associated with decreased patient survival. EGFR and p-EGFR expression was not significantly associated with patient survival.

Conclusion

Activation of AKT1 was associated with poor prognosis in ESCC.     

Elevation of preoperative serum C-reactive protein level is related to poor prognosis in esophageal squamous cell carcinoma

BACKGROUND AND OBJECTIVES: An increased serum C-reactive protein (CRP) level was found in patients with various malignant tumors and was associated with poor prognosis. The aim of this study was to analyze the clinicopathological significance and the prognostic value of preoperative CRP levels in patients with esophageal squamous cell carcinomas. PATIENTS AND METHODS: The preoperative CRP level was measured by enzyme-linked immunosorbent assay (ELISA) in 150 patients with primary esophageal squamous cell carcinomas. All patients underwent radical surgery without any preoperative therapy. The patients were divided into two groups using a cut-off value of 1.0 mg/dl. The pathological classifications of the tumor were examined according to the TNM/UICC classification. The associations between the clinicopathological factors and CRP level were determined. The prognostic value of CRP was determined using Cox's proportional hazards model. RESULTS: Thirty-five patients (23%) showed high CRP levels (more than 1.0 mg/dl). Statistically significant differences in CRP levels were observed depending on tumor depth (P = 0.022) and TNM/UICC stage (P = 0.001). A high CRP level was associated with poor survival (P = 0.005) and was confirmed by multivariate analysis. CONCLUSIONS: A high CRP level is associated with tumor progression and poor survival in patients with esophageal squamous cell carcinoma.     

食管鳞癌组织LSDl表达与预后相关性分析

目的：研究组蛋白赖氨酸特异性脱甲基酶1（1ysinespecificdemethylase1,LSDl）在食管鳞癌组织中的表达及其与临床病理因素的关系,并探讨其与预后的相关性。方法：收集2005—01—01—2006—12—31在福建省肿瘤医院胸外科接受食管癌三野根治术且术前未接受放疗或化疗的食管鳞癌患者135例。免疫组化检测135例食管鳞癌及其配对癌旁正常食管黏膜组织的LSDl表达水平。运用Y。检验分析肿瘤组织LSDl表达与临床病理因素的关系。运用Kaplan-Meier方法和Log—rank检验分析肿瘤组织LSDl表达与患者术后总生存时间的关系。采用Cox模型对食管癌患者预后相关因素进行多因素回归分析。结果：食管鳞癌组织LSDl强阳性表达率为53．3％,在正常食管黏膜组织中为7．6％,差异有统计学意义,x2=9．016,P=0．002。LSDl高表达与性别（x2=0．396,P=0．546）、年龄（x2=2．530,P=0．123）、T分期（x2=1．264,P=0．286）、淋巴结转移（x2=1．136,P=0．343）、TNM分期（x2=0．396,P=0．546）和分化（x2=0．415,P=0．537）无显著相关性,而与生存时间是否超过5年（x2=6．699,P=0．013）显著相关。Cox多因素回归分析显示,淋巴结转移（p=0．001）、肿瘤浸润深度（P=0．004）和LSDl表达水平（P=0．020）是食管鳞癌患者的独立预后因素。生存分析提示,LSDl强阳性组患者预后明显差于LSDl弱阳性组患者,P=0．008。亚组分析显示,在有淋巴结转移的患者中,肿瘤LSDl强阳性与患者预后相关,P=0．014;而在无淋巴结转移的患者中,LSDl强阳性与预后无显著相关性,P=0373结诊．T-Snl在仓管鳞痛钼织申表达上调．其强阳性表达与不良预后相关。     

Increased Numb protein expression predicts poor clinical outcomes in esophageal squamous cell carcinoma patients

Numb is a protein whose asymmetric segregation during cell division determines cell fate and has numerous functions relevant to multiple fields of study, including developmental neurobiology and cancer biology. Little is known about the role of Numb in esophageal squamous cell carcinoma (ESCC), the predominant histological esophageal carcinoma in Asian populations. In this study, we focused on the expression and biologic functions of Numb in the context of ESCC. From analysis of tissue microarrays with 212 patients, it was found that Numb was significantly upregulated in ESCC tissues compared with corresponding non-cancerous tissues. Kaplan-Meier survival analysis suggests that higher expression of Numb was significantly associated with a high tumor recurrence (p = 0.015) and poor overall post-surgical survival (p = 0.016). Using multiple Cox regression, the expression of Numb was determined to be an independent predictor of poor prognosis. When siRNA was used to knockdown Numb in ESCC cell lines, there was a consistent increase in caspase-3 dependent apoptosis and inhibition of cellular proliferation, as well as downregulation of expression of the cancer stem cell markers Oct-4, SOX-2 and Nanog. In addition, downregulated Numb expression was not significantly associated with the migration of ESCC cells. These results indicate that Numb acts as an oncoprotein and has potential as a novel prognostic biomarker and therapeutic target in ESCC patients.     

FGFR1 signaling potentiates tumor growth and predicts poor prognosis in esophageal squamous cell carcinoma patients

Fibroblast growth factor receptor-1 (FGFR1) over-expression was broadly found in squamous cancer, where it induced cellular proliferation, differentiation, and metastasis by activating various signaling pathway. However, the role of FGFR1 gene expression in predicting prognosis of Esophageal Squamous Cell Carcinoma (ESCC) and its regulatory function in the progression of ESCC are not well understood. Therefore, we performed an analysis of FGFR1 mRNA expression by quantitative RT-PCR in tumor tissue of 145 patients with ESCC. The relationships between FGFR1 gene expression and clinicopathological parameters, also the prognosis were further examined. Results suggested that higher FGFR1 gene expression predicted worse overall survival (HR = 1.502, 95%[CI] = 1.005–2.246, P = 0.045). Disease-free survival tends to be shorter in patients with higher FGFR1 expression but without statistical significance (HR = 1.398, 95%[CI] = 0.942–2.074, P = 0.096). FGFR1 was up regulated in multiple ESCC cell lines. Subsequent in vitro experiments demonstrated that anti-FGFR1 treatment by PD173074 inhibited TE-1 and EC9706 cell viability along with the attenuation of MEK-ERK signaling pathway. In vivo, PD173074 administration also had shown potent ESCC growth arresting effect. Overall, our study suggested that FGFR1 gene expression could be an independent prognosis predictive factor in patients with ESCC. Anti-FGFR1 inhibited ESCC growth and could be a potential strategy in ESCC targeted therapy.     

eIF4E在食管鳞状细胞癌中表达及其与预后关系的研究

目的：研究真核细胞翻译起始因子4E（eukaryotic translation initiation factor 4E,eIF4E）在食管癌的表达及其与预后的关系。方法：采用RT-PCR及免疫组化SP法检测食管癌组织及正常食管黏膜组织中eIF4E的表达,分析其表达与食管癌临床病理特征及预后的关系。结果：eIF4E mRNA在食管癌及癌旁组织中相对表达量分别为0.561±0.079,0.442±0.061（P〈0.05）;不同分化的食管鳞癌eIF4E表达量不同（P=0.012）,多重比较显示低分化鳞癌的表达量高于高分化鳞癌的表达（P〈0.05）;不同浸润深度的eIF4E mRNA的表达量不同（P=0.003）,多重比较显示在深肌层和纤维膜的表达量高于黏膜层的表达量（P〈0.05）。eIF4E蛋白在食管癌和正常食管黏膜中的阳性表达率分别为75.00%、30.80%（P=0.001）;与病理学分级呈正相关（rs=0.228,P〈0.05）;不同浸润深度的eIF4E蛋白表达量不同（P〈0.05）。而两者均与食管癌的患者年龄、性别、淋巴结转移和临床分期等临床病理学特征无关（P〉0.05）。eIF4E蛋白高表达组及低表达组5年累积生存率分别为31.8%、53.3%（P〈0.05）。Cox模型分析得出eIF4E蛋白不能作为判断食管鳞癌预后的独立指标（P＝0.211）。结论：eIF4E表达与食管鳞状细胞癌的发生密切相关,其可能参与了肿瘤的浸润和发展,并可以和其他指标共同判断预后。     

Up regulation of the long non-coding RNA NEAT1 promotes esophageal squamous cell carcinoma cell progression and correlates with poor prognosis

Dysregulation of NEAT1 plays critical oncogenic roles and facilitates tumorigenesis on various human tumor entities. However, little information is available about the expression pattern of NEAT1 in esophageal squamous cell carcinoma (ESCC). The contributions of this lncRNA to tumorigenesis and progression of ESCC aslo remains unclear. By performing quantitative real-time polymerase chain reaction (qRT-PCR) in 96 cases of ESCC, we found that the expression of NEAT1 was higher in ESCC tissues and cells compared with the normal counterparts. Pearson analysis showed that elevated NEAT1 levels were extraordinarily correlated with the tumor size (P=0.026), lymph node metastasis (P=0.035) and clinical stage (P=0.004). Moreover, Kaplan–Meier curves with the log-rank test showed that higher expression of NEAT1 led to a significantly poorer survival and multivariate Cox proportional hazards analysis revealed that NEAT1 was an independent risk factor of overall survival (OS). We also assessed the function of NEAT1 in vitro by gain-/loss-of-function studies. Results showed that enhanced expression of NEAT1 stimulated the proliferation of ESCC cells, and promoted their ability of forming foci, migration, and invasion. Conversely, knockdown of NEAT1 showed the opposite effect. Overall, our study indicated that the inappropriate activation of NEAT1 predicts poor prognosis and has a crucial regulatory role in in ESCC. Targeting NEAT1 could be a novel therapeutic choice for treating ESCC patients.     

p16 protein expression is associated with a poor prognosis in squamous cell carcinoma of the lung

An immunohistochemical analysis for p16 protein was performed in 171 patients with non-small-cell lung cancer (NSCLC). Sixty-two carcinomas (36.3%) were classified as p16-negative. p16-negative tumours in squamous cell carcinomas (SCCs) were significantly more than those in adenocarcinomas (P = 0.039). There was no significant difference in survival according to tumour p16 status in patients with NSCLCs or in patients with adenocarcinomas. In contrast, of patients with SCCs, the 5-year survival rate of patients with p16-negative tumours was significantly lower than those with p16-positive tumours (P = 0.001). Especially, the survival of patients with p16-negative tumours was significantly worse than that of patients with p16-positive tumours in the early stage of the SCC, e.g. stage I (P = 0.005). Multivariate analysis showed that p16 status and nodal status were significant prognostic factors for the survival of patients with SCCs of the lung (P = 0.024 and P = 0.008 respectively). In conclusion, our study showed that alteration of p16 was one of the significant factors of a poor prognosis in SCCs of the lung, and that p16 might play an important role in some SCCs of the lung due to its high prevalence and prognostic value.     

RACK1 predicts poor prognosis and regulates progression of esophageal squamous cell carcinoma through its epithelial-mesenchymal transition

Background. RACK1 is known to be involved in tumor progression, and its prognostic value on many kinds of tumors has been identified. However, there are limited studies about the functional role of RACK1 in esophageal squamous cell carcinoma (ESCC). Patients and methods. RACK1 expression was examined in 100 ESCC tissue samples using immunohistochemistry staining. RACK1 was knocked-down in ESCC cell lines by shRNA. The effects on cell proliferation, invasion and migration were examined in ESCC cell lines and nude mouse model. Vimentin and E-cadherin were introduced to further study the association between RACK1 and EMT. Results. RACK1 expression was significantly associated with the tumor length (P = 0.012), diameter<3 cm (P = 0.047), T stage (P = 0.032), and lymph node metastasis (P = 0.038), respectively. Kaplan-Meier survival analysis and Cox analyses revealed RACK1 expression was an independent predictor for OS (P = 0.030) and DFS (P = 0.027) in ESCC. Down-regulation of RACK1 inhibited cell proliferation, along with invasion and migration in vitro and in vivo. A significant positive correlation between RACK1 expression and vimentin (P = 0.0190) and an inverse correlation between RACK1 expression and E-cadherin (P = 0.0047) were found. Conclusions. RACK1 predicted poor prognosis in ESCC, promoted tumor progression, and was involved in EMT of ESCC.     

Spindle cell morphology is related to poor prognosis in vulvar squamous cell carcinoma

Background:

Vulvar cancer is the fourth most common gynaecological malignancy, with an annual incidence of 2 out of 100 000 women. Although most cases of early stage vulvar cancer have a good prognosis, recurrence and rapid tumour progression can occur. We investigated the prevalence of spindle cell morphology in vulvar cancer and its association with survival.

Methods:

This retrospective cohort study included 108 patients with primary vulvar squamous cell carcinoma who were treated at the Leiden University Medical Center during 2000–2009. Paraffin-embedded tissue was examined for the presence of spindle cell morphology. Survival and histology data were compared between cases with spindle and without spindle cell morphology.

Results:

Twenty-two (20%) tumours showed spindle cells infiltrating the stromal tissue. All spindle cell tumours were human papillomavirus (HPV) negative. Spindle cell morphology was strongly associated with poor prognosis and with a high risk of lymph node involvement at the time of diagnosis (relative risk 2.26 (95% CI 1.47–3.47)). Five-year disease-specific survival was lower in patients with vs without spindle cell morphology (45.2% vs 79.7%, respectively; P=0.00057).

Conclusion:

Vulvar spindle cell morphology occurs frequently and seems to develop through the non-HPV pathway. It is associated with a worse prognosis than conventional vulvar squamous cell carcinoma.