Expression of RKIP,E-cadherin and NF-kB p65 in esophageal squamous cell carcinoma and their correlations

To detect the expression of RKIP, E-cadherin and NF-kB p65 in esophageal squamous cell carcinoma (ESCC) and study their correlations. Steptavidin-peroxidase (S-P) method was employed to detect the expressions of RKIP, E-cadherin and NF-kB p65 in ESCC tissues from 77 cases and paracancerous tissues from 77 cases. The correlations between their expressions and clinicopathological indices and between the expressions of these proteins themselves were analyzed. The expressions of RKIP and E-cadherin in ESCC tissues were obviously lower than those in the paracancerous tissues (P<0.01); the expressions in ESCC tissues from cases with lymph node metastasis were lower than those from cases without lymph node metastasis (P<0.01); the expression of RKIP was positively correlated with the expression of E-cadherin in ESCC tissues (P<0.01). The expression of NF-kB p65 in ESCC tissues was correlated with clinical staging, lymph node metastasis and tumor differentiation (P<0.01); the expression of RKIP was negatively correlated with the expression of NF-kB p65 in ESCC tissues (P<0.05). Downregulation or depletion of RKIP was related to the onset and progression of ESCC, and facilitated the invasion and metastasis of ESCC by downregulating E-cadherin and upregulating NF-kB p65.     

Human leukocyte antigen G is associated with esophageal squamous cell carcinoma progression and poor prognosis

Human leukocyte antigen G (HLA-G) is a non-classical HLA class I molecule thought to play a key role in maternal-fetal tolerance and cancer immune evasion. This study aimed to investigate the HLA-G expression in lesion sections and plasma sHLA-G levels of primary esophageal squamous cell carcinoma (ESCC) patients and its clinical significance in diagnosis and prognosis of ESCC. 60 ESCC patients and 28 healthy controls were recruited, and the positive expression of HLA-G in ESCC lesions and adjacent normal tissues were 70% (42/60) and 8.6% (5/60) (P < 0.05), respectively, while no expression was found in normal controls. HLA-G1 and HLA-G5 were determined to be dominating isoforms measured by RT-PCR. There was a significant difference in plasma sHLA-G levels between patients with ESCC (15.04 U/ml, range 4.33–250.00 U/ml) and healthy controls (6.81 U/ml, range 0–29.27 U/ml) (P < 0.01). The plasma IL-10 level was higher in ESCC patients than the controls (23.86 pg/ml vs. 12.81 pg/ml, P < 0.01). HLA-G expression in lesion tissues was correlated with cancer cell differentiation (P = 0.033), lymph node metastasis (P = 0.035) of ESCC. However, no obvious correlations were demonstrated between the plasma sHLA-G levels and the clinicopathological parameters. There was a significant correlation between sHLA-G and IL-10 expression (r = 0.353, P = 0.006) in patients with Esophageal squamous cell carcinoma. HLA-G positive expression showed poorer prognosis of ESCC. HLA-G positive expression might serve as a potential marker in the diagnosis or prediction of ESCC.     

Up-regulation of miR-335 predicts a favorable prognosis in esophageal squamous cell carcinoma

Introduction: MicroRNAs (miRNAs) are noncoding RNAs that regulate multiple cellular processes during cancer progression. MiR-335 has recently been identified to be involved in tumorigenesis of several cancers such as ovarian cancer and gastric cancer. However, the regulation of miR-335 in esophageal squamous cell carcinoma (ESCC) has not been reported yet. Methods: Expression of miR-335 in tumor and their normal matched tissues was determined by quantitative real-time PCR in 67 ESCC patients and its association with overall survival of patients was analyzed by statistical analysis. Results: The expression level of miR-335 was reduced in malignant tissue samples in comparison to normal matched tissue (P < 0.05). It was also proved that miR-335 expression was associated with ESCC histological grade, lymph node metastasis, tumor stage and clinical stage (P < 0.05). In addition, the Kaplan-Meier survival curves revealed that low miR-335 expression was associated with poor prognosis in ESCC patients. Multivariate analysis showed that miR-335 expression was an independent prognostic marker of overall survival of ESCC patients. Conclusions: The study proves for the first time that miR-335 is down regulated in a majority of ESCC patients. Our results indicate that miR-335 expression is an independent prognostic factor for patients with esophageal cancer, which might be a potential valuable biomarker for ESCC.     

Clinicopathologic significance of CXCR4 expressions in patients with esophageal squamous cell carcinoma

AimsThis study was designed to investigate the biological function of CXCR4 in esophageal squamous cell carcinoma and to explore the underlying mechanism to provide potential targets for esophageal squamous cell carcinoma.MethodsA total of 101 patients with esophageal squamous cell carcinoma were included, and the relationship between CXCR4 and clinicopathological factors was analyzed. Laser scanning confocal microscopy was used to observe numbers of autophagosomes in TE-1 cell line and the ability of proliferation and invasion were evaluated meanwhile.ResultsCXCR4 is overexpressed in ESCC specimens and is associated with poor differentiation and lymphocyte metastasis. In the survival analysis, CXCR4 predicted a poor overall survival prognosis. The number of autophagosomes in the siR-CXCR4 group was decreased compared with negative group (P < 0.05), while was increased in the pcDNA3.1-CXCR4 group (P < 0.05).Western blot result show upregulation of LC3II, the ratio of LC3II/LC3I and Beclin1 in pcDNA3.1-CXCR4 group and decreased expression of LC3II, the ratio of LC3II/LC3I and Beclin1 in siR-CXCR4 group. Transwell assay show CXCR4 overexpression promote the invasion of TE-1 cells and was attenuated by autophagy inhibitor 3-Methyladenine.On the contrary, invasion cell numbers decreased in siR-CXCR4 group and was rescued by autophagy inducer Rapamycin.ConclusionCXCR4 is an indicator of poor prognosis for ESCC. CXCR4 promote autophagy and regulate cell invasion through autophagy in ESCC. Our study provides new insights for the treatment of esophageal squamous cell carcinoma and CXCR4 may serve as a therapeutic target for ESCC.     

RANK overexpression as a novel esophageal cancer marker: validated immunohistochemical analysis of two different ethnicities

This study aimed to evaluate the receptor activator of nuclear factor-κB (RANK) expression statuses of esophageal squamous cell carcinoma (ESCC) patients, high-grade intraepithelial neoplasia (HGIN), low-grade intraepithelial neoplasia (LGIN), and normal esophageal tissues (NETs) in Chinese Han and Kazakh ethnic, as well as the correlation of RANK expression with clinicopathological characteristics. RANK immunohistochemical analysis was conducted to investigate the expression of RANK in 113 ESCC, 36 HGIN, 63 LGIN, and 98 NETs from Han ethnic patients and in 196 ESCC and 76 NETs from Kazakh ethnic patients. The associations of RANK expression with ethnic and clinicopathological characteristics were examined using χ²-test. Upregulated RANK expression was detected in both Han and Kazakh ethnic ESCC tissues, compared with NETs (P = 1.11×10^-5, 0.001, respectively). RANK expression was significantly increased during malignant transformation from normal epithelium into LGIN (P = 2.84×10^-7) and HGIN (P = 7.83×10^-6) tissues in Han ethnic patients. The increased expression of RANK also correlated with lymph node metastasis in Kazakh ethnic ESCC patients (P = 0.019). By contrast, no significant correlation existed between RANK expression and clinicopathological characteristics of Han ethnic ESCC patients. Furthermore, Kaplan-Meier survival analysis showed that ESCC patients with higher expression of RANK protein had significantly worse prognosis than ESCC patients with low or no expression (P = 0.001). In conclusion, this study is the first to identify RANK overexpression as a novel esophageal cancer marker in both Kazakh and Han ethnic ESCC patients. The results support the association of RANK with ESCC across ethnicities. In summary, RANK could be a therapeutic target in ESCC patients.     

ADAMTS-6 is a predictor of poor prognosis in patients with esophageal squamous cell carcinoma

Background

A disintegrin and metalloprotease with thrombospondin motif (ADAMTS) enzymes play important roles in cell functions including adhesion, invasion, migration, and proliferation. ADAMTS-6 is a member of the ADAMTS family; reports of its relationship with esophageal squamous cell carcinoma (ESCC) progression are rare. It is unclear whether ADAMTS-6 could be an independent ESCC biomarker.

Significance of elevated ERK expression and its positive correlation with EGFR in Kazakh patients with esophageal squamous cell carcinoma

Extracellular signal-regulated kinases (ERKs) are activated by the MAPK pathway. ERKs are downstream effectors of the epidermal growth factor receptor (EGFR), which belongs to the receptor tyrosine kinases family. Studies on the activation of the EGFR-ERK pathway in Kazakh patients with esophageal squamous cell carcinoma (ESCC) have not been reported. Using immunohistochemical staining on tissue microarrays, we investigated the protein expression of EGFR and ERK in 90 ethnic Kazakh patients with ESCC and 48 adjacent normal esophageal tissues (NETs). EGFR and ERK1 expression was localized in the cytoplasm, whereas ERK2 expression was localized in the nucleus. Both were more highly expression in the ESCC tissues than in the NETs, and the difference was considered significant (P = 0.003, 0.002, and 0.005, respectively). ERK1 and EGFR expression was positively correlated with lymph nodes metastasis (P = 0.011 and 0.013, respectively). ERK1 staining was also significantly associated with tumor-node-metastases stage of ESCC (P = 0.044). ERK2 staining was significantly associated with Histological grade (P = 0.012). Furthermore, ERK1 and EGFR expression in the ESCC tissues were positively correlated (r = 0.413, P < 0.001); EGFR was more highly expressed in the ESCC tissues with high ERK1 expression than in the ESCC tissues with low ERK1 expression (4.95 ± 0.57 vs. 3.21 ± 0.35, P = 0.01). This study is thus far the first to demonstrate the correlation between EGFR overexpression and ERK overexpression in Kazakh patients with ESCC. This correlation suggests that the EGFR-ERK signaling pathway participates in ESCC progression and can thus be used as a prognostic marker.     

Overexpression of CD9 correlates with tumor stage and lymph node metastasis in esophageal squamous cell carcinoma

Objective: Esophageal squamous cell carcinoma (ESCC) is one of the leading causes of cancer deaths worldwide. CD9 has been reported to play a critical role in cell motility, growth and metastasis of multiple cancers. The present study investigated the clinicopathological features of CD9, and its biological characteristics in ESCC. Methods: Fifteen normal esophageal tissue specimens, fifty-three ESCC adjacent tissues and one hundred and four ESCC tissues were included in this study. Using immunohistochemistry (IHC), the expression levels of CD9 were evaluated among different samples. And its clinicopathological parameters and its prognostic factors were analyzed. Western blotting was used to measure CD9 expression and colony formation was performed to determine the effect of CD9 on cell growth in ESCC TE-1 cells. Results: Compared with normal esophageal tissues and tumor adjacent tissues, CD9 expression level is significantly higher in ESCC tissues. CD9 expression correlated with tumor stage (P = 0.022) and lymph node metastasis (P = 0.019) in ESCC patients. Furthermore, the small interfering RNA-mediated silencing of CD9 expression in TE-1 cells resulted in increased proliferation as evidenced by increased colony number and colony size. Conclusion: CD9 expression is upregulated in ESCC tissues and its expression is correlated with tumor stage and lymph node metastasis in ESCC patients. CD9 suppresses the proliferation of TE-1 cells. CD9 may present a potential in tumor progression in ESCC.     

FEZ1基因在食管癌变过程中的作用

目的探讨FEZ1基因在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)组织中的表达及其与患者预后的关系。方法采用Western blot和免疫组化法分析FEZ1基因在ESCC中的表达情况,应用SPSS 13.0软件分析其与临床病理资料及预后的关系。结果 FEZ1蛋白阳性表达在食管癌中的表达与浸润深度、肿瘤大小相关(P<0.05);而与肿瘤组织分化程度、淋巴结转移无关(P>0.05)。正常食管组织中FEZ1的蛋白表达(4.3±0.39)明显高于食管癌组织中(1.6±0.25),且差异有显著性(P<0.05)。FEZ1的高表达提示患者的5年生存率高。结论 FEZ1基因在食管癌的发生、发展过程中起着重要的作用;FEZ1基因在食管癌中可能是候选的抑癌基因。     

Altered expression and localization of desmoglein 3 in esophageal squamous cell carcinoma

Desmoglein 3 (DSG3), a transmembrane cadherin of the desmosomal cell–cell adhesion structure, plays vital roles in the maintenance of normal epithelial tissue architecture. Reports implicating a role for DSG3 expression in cancer are few and contradictory. In this study, immunohistochemical staining was employed to investigate DSG3 expression and subcellular localization in esophageal squamous cell carcinoma (ESCC), and to correlate changes with clinical characteristics. Results indicate that in normal squamous cell epithelia, strong DSG3 immunoreactivity was observed in the Stratum spinosum, and localization occurred only at the cell membrane. In ESCC, DSG3 immunoreactivity displayed an abnormal cytoplasmic localization that was correlated with cell differentiation (P = 0.018). Most strikingly, in 74.1% of the tumors, DSG3 expression was up-regulated and correlated with regional lymph node metastasis (P = 0.036). Moreover, in patients without lymph node metastasis, cytoplasmic localization of DSG3 correlated with poor prognosis (P = 0.044). These results suggest that DSG3 is involved in the development of ESCC and imply that DSG3 overexpression is likely to be an essential contributor to the aggressive features of esophageal cancer.     

A four actin-binding protein signature model for poor prognosis of patients with esophageal squamous cell carcinoma

The actin cytoskeleton is a dynamic structure with actin-binding proteins (ABPs) playing an essential role in the regulation of migration, differentiation and signal transduction in all eukaryotic cells. We examined the relationship between altered expression of four ABPs and clinical parameters in esophageal squamous cell carcinoma (ESCC). To this end, we analyzed 152 formalin-fixed and paraffin-embedded esophageal curative resection specimens by immunohistochemistry for tensin, profilin-1, villin-1 and talin. A molecular predictor model, based on the combined expression of the four proteins, was developed to correlate the expression pattern of the four ABPs with clinical factors and prognosis of ESCC. According to the results, weak significance was found for tensin in lymph node metastasis (P=0.033), and profilin-1 in pTNM stage (P=0.031). However, our four-protein model showed strong correlation with the 5-year overall survival rate (P=0.002). Similarly, Kendall’s tau-b test also showed the relationship between the collective expression pattern of the four ABPs with lymph node metastasis (P=0.005) and pTNM stage (P=0.001). Our results demonstrate that the collective protein expression pattern of four actin-binding proteins could be a biomarker to estimate the prognosis of ESCC patients.     

Clinicopathological significance and angiogenic role of the constitutive phosphorylation of the FOXO1 transcription factor in colorectal cancer

PurposeThis study aimed to evaluate the clinicopathological significance of phospho-forkhead box O1 (pFOXO1) expression and its impact on the angiogenesis of colorectal cancer (CRC).MethodsWe performed immunohistochemistry in 266 human CRC tissues for pFOXO1, and evaluated its cytoplasmic expression, regardless of its nuclear expression. We also investigated the correlation between pFOXO1 expression and clinicopathological characteristics, survival, microvessel density (MVD), and angiogenesis-related molecules in CRC.ResultspFOXO1 was expressed in the cytoplasm of 100 (37.6 %) of the 266 CRC tissues. Furthermore, pFOXO1 expression was significantly correlated with the left colon and rectum, and with vascular invasion, lymph node metastasis, distant metastasis, and higher pTNM stage. However, there was no significant correlation between pFOXO1 expression and other clinicopathological parameters. MVD was significantly higher in pFOXO1-positive tumors than in pFOXO1-negative tumors (P = 0.025). Among the angiogenesis-related molecules examined, pFOXO1 expression was significantly correlated with SIRT1 (P = 0.002) and VEGF expression (P < 0.001), but not with HIF-1α expression. pFOXO1 expression was significantly correlated with poor overall and recurrence-free survival rates (P = 0.001 and P < 0.001, respectively).ConclusionsTaken together, our results showed that the pFOXO1 expression was significantly correlated with aggressive tumor behavior and poor survival rates. Moreover, pFOXO1 expression may affect tumor progression through SIRT1- and VEGF-induced angiogenesis.     

miR-145 expression level in tissue predicts prognosis of patients with esophageal squamous cell carcinoma

BackgroundMicroRNA-145 (miR-145) is commonly down-regulated and has been identified as a tumor-suppressive miRNA in multiple types of cancers, as well as in esophageal squamous cell carcinoma (ESCC). In the present study, the clinical significance and prognostic value were investigated in ESCC.MethodsA total of 126 patients with ESCC who underwent surgery were included in the present study. miR-145 expression was determined using quantitative real-time polymerase chain reaction assay (qRT-PCR) and was further correlated with patients’ clinicopathological parameters. Overall survival was estimated by using Kaplan-Meier method, and univariate analysis was conducted by log-rank test. The Cox proportional hazards model was used in the multivariate analysis.ResultsmiR-145 expression levels in ESCC tissues were significantly decreased compared with the adjacent normal zones (P < 0.001). We observed that the expression level of miR-145 was positively correlated with the tumor differentiation (P = 0.015), lymph node status (P = 0.007), distant metastasis (P = 0.008), and TNM stage (P = 0.033). ESCC patients with low miR-145 expression level had shorter overall survival than those with high miR-145 expression level (log-rank test, P = 0.032). Furthermore, multivariate Cox regression analysis showed that miR-145 expression level was independent factor in predicting the overall survival of ESCC patients (HR = 1.993, 95% CI: 1.277–8.283, P = 0.023).ConclusionsOur findings indicated that miR-145 expression may be a useful prognostic marker that could be used for predicting overall survival of patients with ESCC.     

Elevated expression of UHRF1 predicts unfavorable prognosis for patients with hepatocellular carcinoma

Aims: The present study was designed to evaluate the different expression of ubiquitin-like with PHD and ring finger domains 1 (UHRF1) in hepatocellular carcinoma (HCC) tissues and the adjacent normal tissues, further explore the correlation between UHRF1 expression and the prognosis of HCC patients. Methods: The UHRF1 expression at protein level in HCC tissues and the adjacent normal tissues were measured by high performance liquid chromatography (HPLC). Chi-square test was used to estimate the relationship between UHRF1 expression and clinicopathologic characteristics of HCC patients. The overall survival of HCC patients with diverse expression of UHRF1 was measured by Kaplan-Meier analysis. Cox regression analysis was conducted to judge the prognostic value of UHRF1 in HCC patients. Results: The UHRF1 was over-expressed in HCC tissues compared with the adjacent normal tissues according to the outcome of HPLC (P<0.001). Besides, the UHRF1 expression was tightly related to distant metastasis, cancer area, and HBV (P<0.05), but shared no correlation with gender, cirrhosis, and bilirubin (P>0.05). Patients with high UHRF1 expression had a shorter overall survival time than those with low UHRF1 expression (P<0.001). Cox regression analysis showed that UHRF1 was significantly linked with the prognosis of HCC patients (P=0.002, HR=5.807, 95% CI=1.901-17.742). Conclusion: UHRF1 was over-expressed in HCC tissues compared to the adjacent normal tissues and UHRF1 expression shared significant relevance with distant metastasis, cancer area and HBV. It could be an important and independent prognostic biomarker for HCC patients.     

Increased HIF-1alpha expression in tumor cells and lymphocytes of tumor microenvironments predicts unfavorable survival in esophageal squamous cell carcinoma patients

The expression of hypoxia-induced factor (HIF)-1α is up-regulated in tumor microenvironments under hypoxia condition. However, the prognostic significance of HIF-1α in esophageal squamous cell carcinoma (ESCC) is still elusive. We measured the HIF-1α expression by immunochemistry in tumor specimens from 136 resected ESCC; in the current study, the HIF-1α expression in tumor cells was significantly associated with tumor stage (P = 0.003) and lymph node metastasis (P = 0.006); whereas the HIF-1α expression in tumor-infiltrating lymphocytes (TILs) had no relationship with patients’ clinicopathological parameters. Patients with high HIF-1α expression in tumor cells or in TILs showed worse survival related to those with low HIF-1α expression. Multivariate analysis demonstrated that expression of HIF-1α in TILs was an independent factor for DFS (P = 0.007) and OS (P = 0.013). Additionally, the expression of HIF-1α in tumor cells was an independent factor for DFS (P = 0.037) and OS (P = 0.033) in locoregional ESCC patients, whereas the expression of HIF-1α in TILs was an independent factor for DFS (P = 0.048) and OS (P = 0.039) in metastatic ESCC patients. Correlation analysis revealed that expressions of HIF-1α in tumor cells and in TILs were positively correlated, and patients with combined high HIF-1α in both tumor cells and TILs had the worst survivals (P < 0.05). These findings suggest that the HIF-1α expressions in different cell populations of ESCC microenvironments have different clinical relevance and prognostic impact on patients.     

Nuclear PKM2 expression predicts poor prognosis in patients with esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma (ESCC) is one of the most common tumors worldwide, with a high malignant degree and poor prognosis. The present study aims to investigate the relationship between pyruvate kinase M2 (PKM2) expression and the prognosis of patients with ESCC. The expression of PKM2 in 86 cases of esophageal carcinoma tissues was tested using immunohistochemistry. The relationship between PKM2 expression and clinical pathological parameters, and their effects on the prognosis of patients with ESCC were analyzed. The expression levels of PKM2 in both cytoplasm and nucleus of ESCC tissues were significantly higher than those in paracancerous tissues (P = 6.73 × 10⁻⁹ and 4.32 × 10⁻⁶, respectively). The Kaplan–Meier analysis showed that nuclear PKM2 expression was closely related to the survival of patients with ESCC (P = 0.005). Patients with high PKM2 expression in the nucleus had significantly shorter survival times than those with low PKM2 expression in the nucleus (hazard ratio for death, 2.358; 95% confidence interval, 1.156–4.812; P = 0.018). No other significant difference was found between PMK2 expression and clinico-pathological features of ESCC patients (all P > 0.05). In conclusion, high PKM2 expression in the nucleus is essential in the pathogenic process of ESCC and may be used to predict the prognosis of patients with ESCC.     

microRNA-195-Cdc42 axis acts as a prognostic factor of esophageal squamous cell carcinoma

Background and purpose: Previous studies observed the downregulation of microRNA (miR)-195 in esophageal squamous cell carcinoma (ESCC) tissues, confirmed cell division cycle 42 (Cdc42) as one target gene of miR-195, and demonstrated that miR-195 may act as a tumor suppressor in ESCC by regulating Cdc42 expression. This study aimed to explore the association of miR-195 and Cdc42 combined expression with clinicopathologic factors and prognosis. Methods: Expression of miR-195 and Cdc42 mRNA in 98 pairs of ESCC and paracancerous tissues were detected using real-time quantitative RT-PCR. Results: miR-195 downregulation and Cdc42 upregulation were both prevalent in ESCC tissues, and negatively correlated with each other. In addition, miR-195 expression negatively correlated with TNM stage (P=0.008) and lymphatic metastasis (P=0.022), while Cdc42 expression positively correlated with TNM stage (P=0.011) and tumor differentiation (P=0.024). Moreover, combined expression of miR-195 and Cdc42 (miR-195/Cdc42) was found to be prognostic indicators for progression-free survival and overall survival of ESCC patients both in univariate and multivariate analyses. Conclusion: The main findings of this study indicate the involvement of miR-195-Cdc42 axis in the progression of ESCC and suggest that the combined aberrant expression of miR-195 and Cdc42 mRNA can serve as a promising unfavorable prognostic biomarker in ESCC.