Survival In Esophageal Squamous Cell Carcinoma Research Articles

CD36 Expression Is Associated with Cancer Aggressiveness and Energy Source in Esophageal Squamous Cell Carcinoma.

Esophageal squamous cell carcinoma (ESCC) is an important cause of cancer-related death worldwide. CD36, a long-chain fatty acid (FA) receptor, can initiate metastasis in human oral squamous cell carcinoma (SCC), and its expression is associated with poor prognosis in several cancers. The clinical significance of CD36 expression and its function in ESCC remain unknown. We examined the clinical significance of CD36 expression in 160 ESCC samples using immunohistochemical staining. Functional analysis was performed to determine the association between CD36 and ESCC characteristics (proliferative ability, invasive ability, and energy source dependency). Thirty (18.8%) ESCC cases showed high CD36 expression, indicating a significant association with progression. CD36 suppression inhibited proliferation and invasiveness in ESCC cells. ESCC cells with CD36 suppression used specific essential amino acids (EAAs) as energy sources. Cell viability depended on FAs under CD36 expression. The viability of ESCC cells with CD36 suppression depended on EAAs but not FAs. CD36 may be a good biomarker and therapeutic target in ESCC. Our data provide new insights into the basic mechanism of CD36-dependent energy utilization for ESCC survival. CD36 might be a key regulator of the dependency of FAs as energy source in ESCC cells.

Preoperative Prognostic Nutritional Index is a Significant Predictor of Survival in Esophageal Squamous Cell Carcinoma Patients

Background: Preoperative assessment of patients is meaningful to predict survival in patients with malignant tumors. The prognostic nutritional index (PNI) is one of the most significant factors related to the prognosis in various types of cancer; however, its role in esophageal cancer is still inconclusive. The aim of this study was to identify the prognostic value of PNI in predicting overall survival (OS) in esophageal squamous cell carcinoma (ESCC).Methods: This retrospective study included 4146 ESCC patients, 3812 who underwent esophagectomy for ESCC. Other 334 had no surgery. The Preoperative PNI was measured before any therapies and calculated as 10 × serum albumin (g/dL) + 0.005 × total lymphocyte count (per mm3). We classified the patients into three categories according to the PNI, >50, 45–50, and <45.Results: Our study showed that PNI was associated with age (P<0.0001), gender(P<0.001),tumor length (P<0.0001), T grade (P = 0.001), N staging (P = 0.017),and M staging (P<0.0001). Multivariate analysis showed that PNI was a significant predictor of overall survival Lower PNI vs. Higher PNI group had significantly increased the hazard ratio of ESCC survival (OR = 1.2, 95% CI= 1.05-1.5, p = 0.01). The Kaplan-Meier curve suggested that high PNI group will significantly increase the OS in both surgical and non-surgical group.Conclusion: PNI is a useful predictive factor for long-term survival in ESCC. The survival rate of ESCC can be discriminated between three groups, PNI, >50, 45–50, and <45. The prognostic value of PNI can be applied for both surgical and non-surgical ESCC patients.

MicroRNA-146a as a Prognostic Biomarker for Esophageal Squamous Cell Carcinoma.

Background and AimsMicroRNAs including miR146a have a regulatory role on the expression of genes and act with binding to 3ʹ-UTR region of the genes. Cyclooxygenase-2 (COX-2) is involved in carcinogenesis as an inflammatory marker, and microRNA-146a (miR-146a) as a negative regulatory factor. We aimed to evaluate miR146a expression as a prognostic or diagnostic biomarker for esophageal squamous cell carcinoma (ESCC) and also an association between miR146a and COX2 expression.Materials and MethodsWe quantified the level of miR-146a and COX-2 expression in cancerous and adjacent normal tissue samples obtained from 34 patients with ESCC, using real-time–PCR. Statistical analyses were conducted using one-sample t-test. Receiver-operating characteristic (ROC) curve and Kaplan–Meier analysis were applied to assay miR146a as a diagnostic and prognostic marker, respectively, during 4 years of the study. Furthermore, the Cox regression model was performed to assay the hazard ratio (HR). The association between miR-146a and COX2 expression level in ESCC patients was evaluated by nonparametric Spearman’s rho analysis.ResultsThe results revealed a reduction of miR-146a expression in 50% of cancerous tissue when compared with adjacent normal regions (P-value=0.127). COX-2 expression in 80% of ESCC patients was higher than in the controls (P-value=0.001). Overall, in 60% of cases, direct association was seen between microRNA-146a and COX-2 expression level (correlation coefficient= 0.438, P-value=0.011). COX2 can be considered as a diagnostic biomarker (AUC=0.834, sensitivity=72%, specificity =83%, P-value<0.0001) but miR146a cannot be considered as a diagnostic biomarker (AUC=0.553, sensitivity=88%, specificity =28%, P-value=0.453). Survival analysis by Kaplan–Meier method showed miR146a and COX2 expression can be probably considered as prognostic biomarkers for ESCC because patients with high expression of miR146a had 7 months shorter life span and patients with low expression of COX2 had 8 months shorter life span.ConclusionCOX2 expression is a diagnostic biomarker. MiR-146a and COX2 expression can probably be considered as prognostic biomarkers for survival in ESCC.

A Novel Clinical Six-Flavoprotein-Gene Signature Predicts Prognosis in Esophageal Squamous Cell Carcinoma.

Flavoproteins and their interacting proteins play important roles in mitochondrial electron transport, fatty acid degradation, and redox regulation. However, their clinical significance and function in esophageal squamous cell carcinoma (ESCC) are little known. Here, using survival analysis and machine learning, we mined 179 patient expression profiles with ESCC in GSE53625 from the Gene Expression Omnibus (GEO) database and constructed a signature consisting of two flavoprotein genes (GPD2 and PYROXD2) and four flavoprotein interacting protein genes (CTTN, GGH, SRC, and SYNJ2BP). Kaplan–Meier analysis revealed the signature was significantly associated with the survival of ESCC patients (mean survival time: 26.77 months in the high-risk group vs. 54.97 months in the low-risk group, P < 0.001, n = 179), and time-dependent ROC analysis demonstrated that the six-gene signature had good predictive ability for six-year survival for ESCC (AUC = 0.86, 95% CI: 0.81–0.90). We then validated its prediction performance in an independent set by RT-PCR (mean survival: 15.73 months in the high-risk group vs. 21.1 months in the low-risk group, P=0.032, n = 121). Furthermore, RNAi-mediated knockdown of genes in the flavoprotein signature led to decreased proliferation and migration of ESCC cells. Taken together, CTTN, GGH, GPD2, PYROXD2, SRC, and SYNJ2BP have an important clinical significance for prognosis of ESCC patients, suggesting they are efficient prognostic markers and potential targets for ESCC therapy.

Elevated preoperative platelet distribution width predicts poor prognosis in Esophageal Squamous Cell Carcinoma

Activated platelets play a multifaceted role in tumorigenesis and progression. Platelet distribution width (PDW) is generally applied platelet parameters from routine blood test. Preoperative PDW has been considered a prognostic factor in many cancers. Nevertheless, the prognostic value of PDW in esophageal squamous cell carcinoma (ESCC) remains unknown. The study aimed to investigate whether preoperative PDW could serve as a prognostic factor in patients with ESCC. A total of 495 patients with ESCC undergoing curative surgery were enrolled. The relationship between PDW and clinical features in ESCC was analyzed using chi-square tests. Receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value. Overall survival (OS) and disease-free survival (DFS) stratified by PDW were evaluated by Kaplan–Meier method and log-rank test. Univariate and multivariate Cox regression were used to evaluate the prognostic effect of PDW. Of the 495 patients, elevated PDW was observed in 241(48.7%) of the patients, respectively. An elevated PDW was correlated with depth of tumor (T stage, P = 0.031), nerve infiltration (P = 0.016), hospital time after operation (P = 0.020), platelet (P < 0.001), red cell distribution width (P < 0.001), and aspartate transaminase (P = 0.001). Moreover, elevated PDW (PDW ≥ 13.4 fL) predicted a worse OS and DFS in patients with ESCC (both P < 0.001). Multivariate analyses revealed that PDW was independently associated with OS (hazard ratios 1.194; 95% confidence interval 1.120–1.273; P < 0.001) and DFS (hazard ratios 2.562; 95% confidence interval 1.733–3.786; P < 0.001). Our findings indicated that elevated PDW could serve as an independent worse survival in ESCC.

Serum Fibrinogen Is An Independent Prognostic Factor In Operable Esophageal Squamous Carcinoma: A Real-World Study.

PurposeTo fully elucidate the relationship of serum fibrinogen and the prognosis of esophageal squamous cell carcinoma.MethodsThe concentrations of serum fibrinogen were measured by the Clauss method in a total of 1331 operable esophageal squamous cell carcinoma (ESCC) patients from the past 3 years. The correlation of serum fibrinogen concentration and patient outcome and clinical pathological factors was tested.ResultsHyperfibrinogenemia was significantly correlated with advanced pathological T stage and TNM stage. Serum fibrinogen was determined as a worse independent survival predictor in ESCC by survival analysis. The hazard ratio was 1.672 (95% confidence interval [CI] 1.043–2.681). A subgroup analysis further elucidated a significant correlation between hyperfibrinogenemia and younger age (P=0.011), being female (P=0.002), not having a history of smoking (p<0.001) or alcohol consumption (p<0.001), pathological N0 stage (P=0.002), and early pathological TNM stage (I-II) (P=0.004).ConclusionThis research showed that preoperative serum fibrinogen is an independent prognostic factor for survival in ESCC, especially for the early pathological TNM stage (I-II) and N0 patients.

Analysis of the National Cancer Database Esophageal Squamous Cell Carcinoma in the United States

Esophageal squamous cell carcinoma (ESCC) has been poorly studied, approached with therapeutic nihilism, and likely undertreated. We studied the impact of clinical and patient factors on the survival of ESCC in the United States. We selected patients with stage I to III ESCC from 2004 to 2013, using the National Cancer Database. Patients were categorized into the following treatment modalities: (1) definitive chemoradiation therapy (CR), (2)neoadjuvant therapy followed by esophageal resection (ER), (3) ER alone, and (4) ER followed by adjuvant therapy. Our main outcome measure was overall survival. We identified 11,229 patients with ESCC undergoing definitive CR (78.6%); neoadjuvant therapy followed by ER (8.5%), ER alone (10.1%), and ER followed by adjuvant therapy (2.6%). Compared with neoadjuvant therapy, both ER alone and definitive CR were associated with substantially increased mortality. Patients treated at high-volume centers (>20), regardless of whether they underwent ER, had improved survival compared with facilities that performed 10 to 19, 5 to 9, and less than 5 ERs per year. Patients treated at high-volume facilities were more likely to receive neoadjuvant therapy, and there was a marked inverse relationship between annual surgical volume and long-term survival for both surgically and non-surgically treated patients with stage I to III ESCC.

PRAF2 overexpression predicts poor prognosis and promotes tumorigenesis in esophageal squamous cell carcinoma

BackgroundPrenylated Rab acceptor 1 domain family, member 2 (PRAF2) is involved in the occurrence and progression of several malignant tumors. However, its potential role in esophageal squamous cell carcinoma (ESCC) is still unknown.MethodsPRAF2 mRNA expression was determined in 77 frozen ESCC samples by quantitative reverse transcription-polymerase chain reaction (qPCR) and its association with clinical features and overall survival were evaluated. The roles of PRAF2 in ESCC cells were investigated by proliferation, cell cycle, invasion and apoptosis assays in vitro.ResultsThe PRAF2 mRNA expression was significantly increased in ESCC tissues compared with matched surrounding non-tumor tissues. Survival analysis showed that high PRAF2 mRNA expression was associated with worse overall survival in ESCC patients. Multivariate analysis revealed that PRAF2 (hazard ratio 2.05, 95% CI 1.10–3.85, P = 0.025) emerged as the independent predictor for poor overall survival in ESCC. The in vitro experiments revealed that knockdown of PRAF2 expression blocked cell proliferation, cell cycle progression and cell invasion and induced cell apoptosis in ESCC cells.ConclusionTaken together, our data demonstrate that PRAF2 could be used as a potential prognostic biomarker and represent a potential therapeutic target for ESCC.

Long non-coding RNA LINC00675 inhibits tumorigenesis and EMT via repressing Wnt/β-catenin signaling in esophageal squamous cell carcinoma.

Long noncoding RNA LINC00675 (LINC00675) seems to play an anti-oncogenic role in cancers, though its exact function remains unknown. Up to date, little is known about the role of LINC00675 in esophageal squamous cell carcinoma (ESCC). In this study, we aimed to explore the expression pattern, clinical significance and biological function of LINC00675 in ESCC. RT-PCR was performed to detect the expression levels of LINC00675 in both ESCC tissue and cell lines. The association of LINC00675 expression with clinicopathological factors and prognosis was statistically analyzed. Cell growth was detected by MTT assay and colony formation assay. Cell apoptosis was evaluated with flow cytometry. Migration and invasion ability of ESCC cells were detected wound healing assay and transwell assays. The expressions of EMT-related proteins and Wnt/β-catenin-related proteins by Western blot were investigated. LINC00675 expression was significantly downregulated in both ESCC tissues and cell lines. Decreased LINC00675 expression was correlated with histological grade, lymph nodes metastasis and advanced clinical stage. Furthermore, LINC00675 could serve as an independent predictor for overall survival in ESCC. Importantly, in vitro experiments indicated that that forced LINC00675 expression significantly suppressed inhibited ESCC cell proliferation, colony formation, migration, invasion and EMT, and promoted cell apoptosis through suppressing Wnt/β-catenin signaling pathway. We suggested that LINC00675 acted as a tumor suppressor in ESCC via regulation of Wnt/β-catenin signaling pathway and may be a new prognostic biomarker and potential therapeutic target for ESCC intervention.

Abstract LB-393: LINC01137 functions as a ceRNA promoting SMAD3 signaling and ESCC metastasis

Abstract BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is one of the leading cause of cancer-related death in China and some other parts of the world. Studies have shown that certain long non-coding RNAs (lncRNAs) are involved in cancer development and progression. In this study, we sought to identify lncRNAs that are associated with metastasis of ESCC and to explore their functions and therapeutic implications. METHODS: We performed RNA sequencing and analyzed the association of lncRNAs with ESCC survial time. The significantly associated lncRNAs that were overexpressed in ESCC were selected for further study. Knockdown or overexpression of interest lncRNAs were performed to investigate their effects on ESCC cell invasion and metastasis in vitro and in vivo in mice xengraft models. Biochemical assays such as RNAi, RNA immunoprecipitation, RNA pulldown and chromatin immunoprecipitation assays were used to examine the interactions among lncRNAs, mRNAs and microRNAs and between proteins and DNA. Patient survival time differentiated by RNA expression levels was estimated by Kaplan-Meier plotting and hazard ratios for death were calculated by Cox models. RESULTS: RNA sequencing identified 9 overexpressed lncRNAs were associated with ESCC survival. Among these lncRNAs, LINC01137 had the strongest effects on invasion and metastasis of ESCC cell lines in vitro and in vivo in mice. We found that ESCCwith high expression of LINC01137 had significantly more lymph node metastases compared with ESCC with low expression of LINC01137. Functional analyses demonstrated that LINC01137 may compete with HOXA1 mRNA for miR-608 binding. Higher expression level of LINC01137 in cells may prevent miR-608 from inhibiting HOXA1 translation. We found that HOXA1 is a transcription activator of SMAD2/3 and knockdown of LINC01137 substantilly reduced the SMAD3 signaling. Treatment of ESCC cells overexpressing LINC01137 with Halofuginone, a plant derivative that can inhibit SMAD3 phosphorylation, significantly reduced the abilities of migration and invasion. CONCLUSION: These results suggest that LINC01137 might function as a ceRNA competing with miR-608 for suppressing HOXA1 expression, which promotes SMAD3 signaling to enhance metastasis of ESCC cells. Citation Format: Mingming Shao, Jiahui Chu, Qionghua Cui, Linna Peng, Yingying Luo, Jiang Chang, Wen Tan, Chen Wu, Dongxin Lin. LINC01137 functions as a ceRNA promoting SMAD3 signaling and ESCC metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-393.

Abstract 3405: A mutational signature associated with alcohol consumption and prognostically mutated driver genes in esophageal squamous cell carcinoma

Abstract Aim: Esophageal squamous cell carcinoma (ESCC) is often diagnosed at advanced and incurable stage. Information on driver genes and prognosticators in ESCC remains incomplete. Herein, we elucidated significantly mutated genes (SMGs), mutational signatures, and prognosticators in ESCC. Methods: Three MutSig algorithms (i.e. MutSigCV, MutSigCL and MutSigFN) and “20/20” ratio-metric were employed to identify SMGs. Nonnegative matrix factorization was used to decipher mutational signatures. Kaplan-Meier survival analysis, multivariate Cox and logistic regression models were applied to analyze association between mutational features and clinical parameters. Results: We identified 26 SMGs, including 8 novel (NAV3, TENM3, PTCH1, TGFBR2, RIPK4, PBRM1, USP8 and BAP1) and 18 previously reported. Three mutational signatures were identified to be prevalent in ESCC including clocklike C&amp;gt;T at CpG, APOBEC overactive C&amp;gt;T at TpCp[A/T], and a signature featured by T&amp;gt;C substitution. The T&amp;gt;C mutational signature was significantly correlated with alcohol consumption (OR: 3.59; 95% CI: 2.30-5.67; P &amp;lt; 0.001). This alcohol consumption signature was also observed in liver cancer and head and neck squamous cell carcinoma, and its mutational activity was substantially higher in samples with mutations in TP53. Survival analysis revealed that TENM3 mutations (HR: 5.54; CI: 2.68-11.45; P &amp;lt; 0.001) and TP53 hotspot mutation p.R213* (HR: 3.37; CI: 1.73-8.06; P &amp;lt; 0.001) were significantly associated with shortened survival outcome. The association remained statistically significant after controlling for age, gender, TNM stage and tumor grade. Conclusions: We have uncovered several new SMGs in ESCC and defined an alcohol consumption related mutational signature. TENM3 mutation and TP53 hotspot mutation p.R213* are independent prognosticators for poor survival in ESCC. Citation Format: Xiangchun Li, Mengyao Wang, Meng Yang, Hongji Dai, Baifeng Zhang, Wei Wang, Xinlei Chu, Xin Wang, Hong Zheng, Ruifang Niu, Wei Zhang, Kexin Chen. A mutational signature associated with alcohol consumption and prognostically mutated driver genes in esophageal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3405.

Prognostic significance of red cell distribution width in esophageal squamous cell carcinoma

BackgroundRed cell distribution width (RDW) is routinely assessed as part of the complete blood count (CBC) to gather information on the heterogeneity in the size of circulating erythrocytes. RDW is a more sensitive screening marker for anemia, inflammation, and nutritional deficiencies. The purpose of this study was to explore the prognostic value of RDW in esophageal squamous cell carcinoma (ESCC) patients. MethodsWe conducted a retrospective study of data from 148 ESCC consecutive patients who underwent potentially curative esophagectomy and analyzed the correlation of RDW with various clinicopathological factors. ResultsMultivariate analyses identified a high RDW (HR, 2.061; P = 0.0286) as a significant risk factor for cancer-specific survival (CSS). Kaplan–Meier analysis and the log-rank test demonstrated that patients with a high RDW had a significantly worse prognosis in terms of CSS than those with a low RDW (P = 0.0011). In multivariate analysis, there was no significant relationship between RDW and CSS in pathological tumor node metastasis stage I or II patients. However, a high RDW (HR, 2.386; P = 0.0471) was confirmed to be an independent worse prognostic factor for CSS in pathological tumor node metastasis stage III cancer patients. Kaplan–Meier analysis and the log-rank test showed a significant relationship between RDW and CSS in patients with pathological tumor node metastasis stage III (P = 0.0175). ConclusionsThe RDW was a significant and independent predictor of poor survival in ESCC patients after curative esophagectomy. RDW may aid clinicians in detecting signs of recurrence very early and effectively customize treatment regimens. RDW is thus a convenient, cost-effective, and readily available biomarker to predict survival in ESCC.

Predicting prognosis in resected esophageal squamous cell carcinoma using a clinical nomogram and recursive partitioning analysis

BackgroundDevelopment demand of precise medicine in resectable esophageal squamous cell carcinoma (ESCC) require to recognize patients at high risk treated by surgery alone. Thus, our aim was to construct a clinical nomogram and recursive partitioning analysis (RPA) to predict long-term survival in ESCC treated by surgery alone. MethodsBased on the patients with ESCC who treated by three-incisional esophagectomy and two-field lymphadenectomy alone, we identified and integrated significant prognostic factors for survival to build a nomogram. The nomogram was calibrated for overall survival (OS) and the predictive accuracy and discriminative ability was measured by concordance index (c-index) and Akaike information criterion (AIC). Based on the nomogram, the RPA was performed for risk stratification. ResultsA total of 747 patients were included for analysis. Five independent prognostic factors were identified and entered into the nomogram. The calibration curves for probability of 1-, 3-, and 5-year OS showed optimal agreement between nomogram prediction and actual observation. The AIC value of the nomogram was lower than that of the 7th edition staging system, whereas the c-index of the nomogram was higher than that of the 7th edition staging system. The risk groups stratified by RPA allowed significant distinction between survival curves within respective TNM categories. ConclusionThe RPA based on a clinical nomogram appears to be suitable for risk stratification in OS for resected ESCC. This practical system may help clinicians in decision making and design of clinical studies.

Survival based radiographic-grouping for esophageal squamous cell carcinoma may impact clinical T stage.

Most patients diagnosed with thoracic esophageal squamous cell carcinoma (ESCC) have progressed beyond surgical resection as a therapeutic option. Difficulties in the proper assessment of tumor invasion depth before treatment complicate determination of the type and extent of therapy. Therefore, accurate tumor clinical staging is a necessity for identifying treatment options and aiding in patient prognosis. We investigated radiographic factors as prognostic indicators for survival in ESCC. Between July 2006 - July 2010, 324 thoracic ESCC patients who underwent surgery were selected. All patients received contrast enhanced preoperative chest CT scans and esophageal barium swallow examinations. Measurement of maximal lesion cross-sectional area, the largest long diameter, largest short diameter, CT-indicated lesion length, barium-indicated lesion length and the length of pericardial fat reduction were performed. Relationships between these indicators and post-surgical survival time and the cutoff values of related factors were analyzed. Maximum long diameter, maximum lesion area and lesion length, as measured by CT imaging, were correlated with survival. Survival effects were clearly associated with group intervals, calculated by a genetic algorithm, and tumor stages. Risk-stratification intervals of esophageal lesions from radiographic imaging included: maximum long diameter < 28.7, 28.7-34.6mm, 34.6-41.4mm and >41.4mm; maximum lesion area < 355.8mm2, 355.8-568.0mm2, 568.0-907.3mm2 and >907.3mm2; and CT-indicated lesion length <30.9mm, 30.9-57.3mm, 57.3-70.6mm and > 70.6mm. The reasonable stratification of maximum esophageal lesion area, largest long diameter and lesion length measured in CT is valuable for clinical T staging of ESCC. Radiographic parameters may have prognostic clinical value in the staging of esophageal carcinoma.

Expression of proteins associated with epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

Cluster of differentiation 147 (CD147), pituitary tumor transforming gene (PTTG) and CD44v6 are proteins involved in the epithelial-mesenchymal transition (EMT). To investigate the prognostic value of CD147 and PTTG, and CD44v6 expression in esophageal squamous cell carcinoma (ESCC), tissue microarray specimens from 76 patients with ESCC were evaluated by immunohistochemistry staining and scored by intensity and proportion of positive areas. Expression levels of CD147, PTTG and CD44v6 were higher in tumor tissues than in matched adjacent tissues. CD147 expression was positively associated with lymph node metastasis (P=0.025) and American Joint Committee on Cancer (AJCC) system clinical grades (P=0.037). CD147 expression was positively correlated with the expression levels of PTTG (R=0.369; P=0.001) and CD44v6 (R=0.320; P=0.005). In addition, Kaplan-Meier analysis indicated that positive expression of CD147, PTTG and CD44v6 was significantly associated with poor overall survival times (P=0.045, P=0.014 and P=0.027, respectively). Patients exhibiting CD147-PTTG co-expression, CD147-CD44v6 co-expression and CD147-PTTG-CD44v6 triple-positive expression had the poorest overall survival rates. In conclusion, the expression of EMT-associated proteins, including CD147, PTTG and CD44v6, was significantly associated with poor survival in ESCC and these novel targets may serve as potential biomarkers for anticancer therapies.

Programmed death-1 ligand-1 gene rs2890658 polymorphism associated with the risk of esophageal squamous cell carcinoma in smokers

The most important anti-tumor immune response is mediated by T lymphocytes. The interaction of programmed death-1 ligand-1 (PD-L1) with its receptor provides an inhibitory signal in T lymphocytes activation and proliferation. This study aimed to investigate whether polymorphisms of PD-L1 were associated with the risk and prognosis of esophageal squamous cell carcinoma (ESCC) in a high-incidence population from Northern China. PD-L1 rs2890658 A/C and rs4143815 C/G single nucleotide polymorphisms (SNPs) were genotyped by polymerase chain reaction ligase detection reaction (PCR-LDR) method in 575 ESCC patients and 577 healthy controls. There was no significant difference in the genotype frequencies of these two SNPs between the ESCC patients and the healthy controls. However, for rs2890658 A/C SNP, compared with the C/C genotype, the A/C genotype increased the risk of ESCC for the smokers (OR = 1.513, 95% CI = 1.006-2.287). Among the 575 ESCC patients, the survival information of 202 ESCC patients was collected. Neither the rs2890658 A/C SNP nor the rs4143815 C/G SNP was associated with the survival of ESCC patients. PD-L1 rs2890658 A/C SNP might be used as risk marker of the susceptibility to ESCC for the Han nationality in a high-incidence population from Northern China.

Macrophage migration inhibitory factor promotes tumor aggressiveness of esophageal squamous cell carcinoma via activation of Akt and inactivation of GSK3β

The pleiotropic pro-inflammatory cytokine, macrophage migration inhibitory factor (MIF), represents an important link between chronic inflammation and tumorigenesis. Although accumulating evidence demonstrates that MIF overexpression is implicated in the development and progression of multiple cancers, including esophageal squamous cell carcinoma (ESCC), the molecular mechanisms underlying its tumor-promoting roles in ESCC remain unclear. In the present study, we observed that MIF is overexpressed in ESCC and correlated significantly with lymph node metastasis, advanced clinical stage, and poor survival of ESCC. MIF knockdown attenuated the proliferation, migration, and invasion of ESCC cells in vitro and in vivo. Moreover, blockage of MIF expression decreased the activation of the Akt, MEK/ERK, and NF-κB pathways and enhanced sensitivity to apoptosis. Meanwhile, repression of MIF expression resulted in activation of glycogen synthase kinase 3 beta (GSK3β) and subsequent decrease of active β-catenin, as well as its downstream targets including cyclin D1, matrix metalloproteinase (MMP)-7, c-myc, and c-Jun. Collectively, our results provided mechanistic insights into the tumor-promoting role of MIF in ESCC, and suggested that MIF represents a potential therapeutic target for treatment of ESCC.

MiR-323a-3p Inhibits Cell Proliferation, Migration, and Invasion in Esophageal Squamous Cell Carcinoma Via Suppression FMR1

Esophageal squamous cell carcinoma (ESCC) is one of the most common and deadly cancers in the world. Currently, the five-year survival rate of ESCC remains poor. Studies show that miRNAs plays an important role in various tumor processes. Our study aimed to discover prognostic miRNA and investigate its role in ESCC. Prognosis related miRNA was detected by ABI TaqMan miRNA chip method using serum of 15 ESCC patients with T3-4/N1, M0-1 stage. The cell proliferation was evaluated by Cell Titer 96 AQueous One Solution Cell Proliferation Assay (MTS), and the migration/invasion was evaluated by transwell assay. The bioinformatics methods were applied to predict the targets of miRNA. The mRNA and protein levels of miRNA target were determined by quantitative real time polymerase chain reaction (qRT-PCR) analysis, western blot and enzyme-linked immunosorbent assay (ELISA). Four weeks old NU/NU nude mice were used for the in vivo tumorigenicity study, including subcutaneous injection for tumor volume evaluation and tail vein injection for metastasis observation. Downregulated miR-323a-3p was closely correlated with the shorter survival of ESCC. In most of ESCC cell lines, such as YSE2, KYSE 150, KYSE 510, the expressions of miR-323a-3p were largely lower than esophageal epithelial cells. MiR-323a-3p repressed cell proliferation of KYSE 30 and KYSE 150. Moreover, miR-323a-3p inhibited cell migration and invasion of YES 2 and KYSE 30. These results further confirmed the tumor suppressor status for miR-323a-3p. Through biological predictions, FMR1 was one of the possible targets of miR-323a-3p. The expression of this miRNA and FMR1 showed negative correlation. The compromised cell migration and invasion induced by miR-323a-3p mimic transfection were rescued by transfection of FMR1 expression plasmid in both YSE2 and KYSE 30. In vivo, tumors induced by KYSE 30-miR-323a-3p grew significantly slower and resulted in smaller tumor mass. Metastatic lung colonization could be inhibited by overexpression of miR-323a-3p in KYSE 30. MiR-323a-3p acts as a tumor suppressor by targeting FMR1. The overexpression of miR-323-3p might offer a potential therapeutic target against metastasis in patients with ESCC.

DNA hypermethyation and silencing of PITX1 correlated with advanced stage and poor postoperative prognosis of esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is associated with the accumulation of genetic and epigenetic changes in the background mucosa. Dysregulated DNA methylation is known to lead to the inactivation of tumor suppressor genes and the activation of oncogenes. To identify the genes whose expression is perturbed by abnormal DNA methylation in ESCC, integrative transcriptomics by serial analysis of gene expression (SAGE) and methylome sequencing by methyl-DNA immunoprecipitation (MeDIP) analysis were performed. We found 159 genes with significantly decreased expression in ESCC compared to that in noncancerous esophageal mucosa. MeDIP-seq analysis identified hypermethylation in the promoter region of 56 of these genes. Using surgically resected tissues of 40 cases, we confirmed that the paired-like homeodomain 1 (PITX1) gene was hypermethylated in ESCC compared to that in normal tissues (P < 0.0001) by pyrosequencing. PITX1 overexpression in ESCC cell lines inhibited cell growth and colony formation, whereas PITX1 knockdown accelerated cell growth. A PITX1-transfected ESCC cell line, KYSE30, formed smaller tumors in nude mice than in mock-transfected cells. Hypermethylation of PITX1 was associated with tumor depth (P = 0.0011) and advanced tumor stage (P = 0.0052) and predicted poor survival in ESCC (hazard ratio, 0.1538; 95% confidence interval, 0.03159–0.7488; P = 0.0169). In this study, we found a novel tumor suppressor gene of ESCC, PITX1, which is silenced by DNA hypermethylation. Downregulation of PITX1 contributes to the growth and progression of ESCC. Hypermethylation of the PITX1 in ESCC correlated with tumor progression and advanced stage cancer, and may predict a poor prognosis.

The overexpression and predictive significance of MMP-12 in esophageal squamous cell carcinoma

Matrix metalloproteinase (MMP)-mediated degradation of the extracellular matrix is a major factor for tumor invasion and metastasis. MMP-12, as metalloelastase, its function in tumor progression remains contradictory. This study was undertaken to investigate the role of MMP-12 in esophageal squamous cell carcinoma (ESCC). We analyzed the protein expression of MMP-12 and its association with clinicopatholigical parameters, as well as survival analysis. MMP-12 was highly expressed in tumor cells comparing with normal epithelial cells. The high expression of MMP-12 was significantly correlated with tumor grade and stage, nodal metastasis and poor survival of ESCC. Cox multivariate analysis revealed that MMP-12 was an independent prognostic factor in ESCC. Our results suggest that MMP-12 might act as a potential target for the development of novel therapeutics of esophageal squamous cell carcinoma.