Stage Esophageal Squamous Cell Carcinoma Research Articles

Low-dose spectral insufflation computed tomography protocol preoperatively optimized for T stage esophageal cancer - preliminary research experience.

AIMTo evaluate the T stage of esophageal squamous cell carcinoma (ESCC) using preoperative low-dose esophageal insufflation computed tomography (EICT).METHODSOne hundred and twenty ESCC patients confirmed by surgery or esophagoscopy were divided into three groups. Groups B and C were injected with 300 mgI/kg contrast medium for automatic spectral imaging assist (GSI assist), while group A underwent a conventional 120 kVp computed tomography (CT) scan with a 450 mgI/kg contrast medium injection. EICT was performed in group C. Group A was reconstructed with filtered back projection, and groups B and C were reconstructed with 50% adaptive statistical iterative reconstruction. The contrast-to-noise ratio of lesion-to-mediastinal adipose tissue and the radiation dose were measured. Specific imaging features were observed, and T stage ESCCs were evaluated.RESULTSThe sensitivity and accuracy of the T1/2 stage were higher in group C than in groups A and B (sensitivity: 43.75% vs 31.82% and 33.33%; accuracy: 54.29% vs 46.67% and 52.50%, respectively). With regard to the T3 stage, the sensitivity and specificity in group C were higher than those in groups A and B (sensitivity: 56.25% vs 41.17% and 44.44%; specificity: 73.68% vs 67.86% and 63.64%, respectively). The diagnostic sensitivity, specificity and accuracy of the T4 stage were similar among all groups. There were no significant differences in volume CT dose index [(5.91 ± 2.57) mGy vs (3.24 ± 1.20) vs (3.65 ± 1.77) mGy], dose-length product [(167.10 ± 99.08) mGy•cm vs (113.24 ± 54.46) mGy•cm vs (117.98 ± 32.32) mGy•cm] and effective dose [(2.52 ± 1.39) vs (1.63 ± 0.76) vs (1.73 ± 0.44) mSv] among the groups (P > 0.05). However, groups B and C received similar effective doses but lower iodine loads than group A [(300 vs 450) mgI/kg].CONCLUSIONEICT combined with GSI assist allows differential diagnosis between the T1/2 and T3 stages. The ability to differentially diagnose the T3 and T4 stages of medullary ESCC can be improved by quantitatively and qualitatively analyzing the adipose tissue in front of the vertebral body.

Combined detection of IL-6 and IL-8 is beneficial to the diagnosis of early stage esophageal squamous cell cancer: a preliminary study based on the screening of serum markers using protein chips.

BackgroundThe diagnosis rate of early stage esophageal squamous cell carcinoma (ESCC) is low due to the lack of specific tumor markers. Seeking for these markers is beneficial to improve the early diagnosis rate and the prognosis of patients. This study profiles the differentially expressed proteins of early stage ESCC patients via the AAH-BLG-507 protein chip, which further consolidates the clinical evidence of ESCC diagnosis.Materials and methodsIn this study, 20 serum samples were collected from Taihe Hospital between August 2016 and June 2017. Ten of them carried ESCC, while the rest were healthy controls. To profile the proteins’ expression level, the AAH-BLG-507 protein chip was used, and both highly expressed and lowly expressed proteins were fished out. Meanwhile, their biological roles were examined by using Gene Ontology (GO) database and String database, and they were further verified by ELISA.ResultsResults showed that the expression levels of AXL, ARTN, Ang2, BDNF, BMP7, cripto-1, CCL28, E-selectin, IL-6, IL-8 and SHH in the serum of early ESCC were significantly upregulated (P<0.05), particularly IL-6 and IL-8. The expression levels of TSP1 and MMP-8 were markedly downregulated (P<0.05). Analysis showed that these proteins were mainly involved in angiogenesis, signal transduction, cell proliferation and migration, indicating the close relationship with the development of ESCC.ConclusionIt suggested that IL-6 and IL-8 proteins could be considered as the markers for ESCC diagnosis.

PS02.066: ROLE OF 18F-FDG-PET/CT IN ESOPHAGEAL SQUAMOUS CELL CARCINOMA AFTER NEOADJUVANT CHEMORADIOTHERAPY

Abstract Background The aim of this study was to assess the role of 18F-fluorodeoxyglucose positron-emission tomography/computed tomography (FDG-PET/CT) in predicting pathological response and survival in patients with esophageal squamous cell carcinoma (ESCC) treated with neoadjuvant chemoradiotherapy (nCRT). Methods Thirty patients with advanced ESCC received nCRT followed by surgery, and underwent FDG-PET/CT twice before and after nCRT. We compared the results of FDG-PET/CT with the pathological results and prognosis. Results Pathological response was found to correlate with the maximum standardised uptake value (SUVmax) after nCRT and the rate of decrease of SUVmax. Using univariate analysis, pN, SUVmax after nCRT and the rate of decrease of SUVmax were found to be prognostic factors. Multivariate analysis revealed that only pN was an independent prognostic factor Conclusion The prediction of pathological response and prognosis using FDG-PET/CT is not as reliable as pathological detection of lymph node metastasis, but could be a useful method contributing to treatment decisions. Neoadjuvant chemoradiotherapy (nCRT) plus surgery has been shown to improve survival rates and should be regarded as a standard of care for patients with locally advanced esophageal squamous cell carcinoma (ESCC). Appropriate evaluation of nCRT efficacy based on noninvasive parameters might help in individualizing treatments for patients with ESCC. 18F-fluorodeoxyglucose positronemission tomography/computed tomography (FDG-PET/CT) reflects tumor cell viability based on enhanced FDG uptake as a result of increased glucose metabolism. FDG-PET/CT is useful for the staging of advanced ESCC before treatment, and for evaluating the response to nCRT; however, findings from currently available studies in this regard are controversial. In the present study, we examined the role and usefulness of FDG-PET/CT in decisions regarding staging, prediction of histopathological response, and overall survival in patients with advanced ESCC treated with nCRT; this was achieved by analyzing the maximum standardized uptake value (SUVmax) before and after treatment, and the rate of decrease of SUVmax. Disclosure All authors have declared no conflicts of interest.

PS02.097: PROGNOSTIC IMPACT OF LYMPH NODE METASTASIS STATIONS IN PN1 STAGE ESOPHAGEAL SQUAMOUS CARCINOMA PATIENTS

Abstract Background Although numerically based lymph node staging system of UICC/AJCC of esophageal squamous cell carcinoma (ESCC) offers excellent prognostic performance, it doesn’t offer information on the anatomical extent of disease. The aim was to evaluate prognostic impact of lymph node metastasis (LNM) stations and regions in pN1 stage patients. Methods Among patients with ESCC undergone esophagectomy classified as R0 from January 2005 to December 2010, 185 patients diagnosed as pN1 were selected for analysis with univariate and multivariate Cox regression analysis. Results The most common metastasis areas in pN1 patients were middle and lower thoracic paraesophageal LN (28.6%), left gastric artery LN (27.6%) and paracardial LN (24.3%). Non- middle and lower paraesophageal LNM (P = 0.049), subcarinal LNM (P = 0.041) and multiple left gastric artery LNM (P = 0.010) were independent risk factors for worse survival. The median survival of the N1 (JES) group was significantly better than the N2 (JES) (40.1 months vs. 22.4 months, P = 0.012). However, no significant difference was found in median survival rate among intra-thoracic LNM, upper-abdominal LNM and both LNM. Conclusion Paraesophageal LNM, subcarinal LNM and left gastric artery LNM are predictive factors for survival and should be swept routinely in pN1 stage ESCC patients. On the basis of UICC/AJCC lymph node Classification system, anatomical lymphatic spread pattern analysis could further improve prognosis prediction. Disclosure All authors have declared no conflicts of interest.

The chromosome 11q13.3 amplification associated lymph node metastasis is driven by miR-548k through modulating tumor microenvironment

BackgroundThe prognosis for esophageal squamous cell carcinoma (ESCC) patients with lymph node metastasis (LNM) is still dismal. Elucidation of the LNM associated genomic alteration and underlying molecular mechanisms may provide clinical therapeutic strategies for ESCC treatment.MethodsJoint analysis of ESCC sequencing data were conducted to comprehensively survey SCNAs and identify driver genes which significantly associated with LNM. The roles of miR-548k in lymphangiogensis and lymphatic metastasis were validated both in vitro and in vivo. ESCC tissue and blood samples were analyzed for association between miR-548k expression and patient clinicopathological features and prognosis and diagnosis.ResultsIn the pooled cohort of 314 ESCC patients, we found 76 significant focused regions including 43 amplifications and 33 deletions. Clinical implication analysis revealed a panel of genes associated with LNM with the most frequently amplified gene being MIR548K harbored in the 11q13.3 amplicon. Overexpression of miR-548k remarkably promotes lymphangiogenesis and lymphatic metastasis in vitro and in vivo. Furthermore, we demonstrated that miR-548k modulating the tumor microenvironment by promoting VEGFC secretion and stimulating lymphangiogenesis through ADAMTS1/VEGFC/VEGFR3 pathways, while promoting metastasis by regulating KLF10/EGFR axis. Importantly, we found that serum miR-548k and VEGFC of early stage ESCC patients were significantly higher than that in healthy donators, suggesting a promising application of miR-548k and VEGFC as biomarkers in early diagnosis of ESCC.ConclusionsOur study comprehensively characterized SCNAs in ESCC and highlighted the crucial role of miR-548k in promoting lymphatic metastasis, which might be employed as a new diagnostic and prognostic marker for ESCC.

Circulating plasma microRNAs in the detection of esophageal squamous cell carcinoma.

Circulating microRNAs (miRNAs/miRs) have been reported as diagnostic biomarkers for esophageal cancer (EC) diagnosis. However, contrasting results have been achieved in different studies. In the present study, a meta-analysis was performed, based on the systematic search of PubMed and Web of Science, to evaluate the diagnostic value of circulating miRNAs in the peripheral blood in EC. The top 5 most-studied miRNAs were selected for confirmation by reverse transcription quantitative-polymerase chain reaction using the blood plasma of 125 patients with esophageal squamous cell carcinoma (ESCC) and 125 healthy individuals from Henan, China. A total of 45 studies from 22 articles, regarding 33 miRNAs were considered in the meta-analysis. The pooled sensitivity and specificity were both 0.79 (95% confidence interval, 0.76-0.82 for both). Among the 5 miRNAs considered (miR-21, miR-223, miR-375, miR-25 and miR-100), miR-21 and miR-223 were significantly overexpressed whereas miR-375 expression was reduced in patients with ESCC compared with healthy individuals (all P<0.001). The areas under the curves (AUCs) were 0.80, 0.73, and 0.69 for miR-21, miR-223, and miR-375, respectively. The AUCs increased when discriminating between patients with early ESCC in stage 0-I and the non-invasive carcinoma stage Tis-T1 stage from controls. Thus, it was concluded that plasma miR-21, miR-223 and miR-375 may serve as non-invasive diagnostic biomarkers in patients with ESCC, especially early ESCC in stages 0-I and Tis-T1.

Practical value of identifying circulating tumor cells to evaluate esophageal squamous cell carcinoma staging and treatment efficacy.

BackgroundThis study was conducted to investigate the correlation between clinicopathological features and post‐therapeutic response in esophageal squamous cell carcinoma (ESCC) patients. Peripheral blood circulating tumor cells (CTCs) expressing epithelial‐mesenchymal transition markers were identified.MethodsPeripheral blood samples were collected from 71 patients with newly diagnosed ESCC and 40 healthy volunteers. CTCs were isolated using CanPatrol CTC enrichment technology. RNA‐fluorescent in situ hybridization was used to phenotype the CTCs on the basis of epithelial and/or mesenchymal markers.ResultsThe median mesenchymal CTC counts in 71 patients were: 0 in 19 stage I patients, 2 in 31 stage II, and 3 in 21 stage III/IV. The overall diagnostic performance of total CTCs to correctly identify ESCC patients was 0.991. We observed a correlation between increases in tumor size or advanced stage and an increased number of mesenchymal CTCs (P < 0.05). Thirty‐nine patients were administered two cycles of neoadjuvant chemotherapy and their therapeutic response was evaluated: 2 complete response, 20 partial response, 13 stable disease, and 4 progressive disease. After treatment, the positive rate of mesenchymal CTCs was 70.6% in the progressive and stable disease group versus 36.4% in the complete and partial response group (P = 0.05).ConclusionThe results showed that mesenchymal CTC count is related to ESCC clinical stage and the efficacy of neoadjuvant chemotherapy.

A novel rapid quantitative method reveals stathmin-1 as a promising marker for esophageal squamous cell carcinoma.

Stathmin‐1 is a microtubule depolymerization protein that regulates cell division, growth, migration, and invasion. Overexpression of stathmin‐1 has been observed to be associated with metastasis, poor prognosis, and chemoresistance in various human cancers. Our previous studies found that serum stathmin‐1 was significantly elevated in patients with esophageal squamous cell carcinoma (ESCC) by ELISAs. Here, we constructed high‐affinity monoclonal antibodies and then developed a competitive AlphaLISA for rapid, accurate quantitation of stathmin‐1 in serum. Compared to ELISA, our homogeneous AlphaLISA showed better sensitivity and accuracy, a lower limit of detection, and a wider linear range. The measurements of nearly 1000 clinical samples showed that serum stathmin‐1 level increased dramatically in patients with squamous cell carcinoma (SCC), especially in ESCC, with a sensitivity and a specificity of 81% and 94%, respectively. Even for early stage ESCC, stathmin‐1 achieved an area under the receiver operating characteristic curve (AUC) of 0.88. Meanwhile, raised concentrations of stathmin‐1 were associated with lymph node metastasis and advanced cancer stage. Notably, various types of SCC showed significantly higher AUCs in serum stathmin‐1 detection compared to adenocarcinoma. Furthermore, we confirmed that stathmin‐1 was enriched in the oncogenic exosomes, which can explain the reason why it enters into the blood to serve as a tumor surrogate. In conclusion, this large‐scale and systematic study of serum stathmin‐1 measured by our newly established AlphaLISA showed that stathmin‐1 is a very promising diagnostic and predictive marker for SCC in the clinic, especially for ESCC.

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.

Preoperative serum immunoglobulin G and A antibodies to Porphyromonas gingivalis are potential serum biomarkers for the diagnosis and prognosis of esophageal squamous cell carcinoma

BackgroundThe key-stone-pathogen, Porphyromonas gingivalis associates not only with periodontal diseases but with a variety of other chronic diseases such as cancer. We previously reported an association between the presence of Porphyromonas gingivalis in esophageal squamous cell carcinoma (ESCC) and its progression. We now report the diagnostic and prognostic potential of serum immunoglobulin G and A antibodies (IgG/A) against Porphyromonas gingivalis for ESCC.MethodsAn enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of Porphyromonas gingivalis IgG and IgA in 96 cases with ESCC, 50 cases with esophagitis and 80 healthy controls.ResultsThe median serum levels of IgG and IgA for P. gingivalis were significantly higher in ESCC patients than non-ESCC controls. P. gingivalis IgG and IgA in serum demonstrated sensitivities/specificities of 29.17%/96.90% and 52.10%/70.81%, respectively, and combination of IgG and IgA produced a sensitivity/specificity of 68.75%/68.46%. The diagnostic performance of serum P. gingivalis IgA for early ESCC was superior to that of IgG (54.54% vs. 20.45%). Furthermore, high serum levels of P. gingivalis IgG or IgA were associated with worse prognosis of ESCC patients, in particular for patients with stage 0-IIor negative lymphnode metastasis, and ESCC patients with high levels of both IgG and IgA had the worst prognosis. Multivariate analysis revealed that lymph node status, IgG and IgA were independent prognostic factors.ConclusionsThe IgG and IgA for P. gingivalis are potential serum biomarkers for ESCC and combination of IgG and IgA improves the diagnostic and prognostic performance. Furthermore, serum P. gingivalis IgG and IgA can detect early stage ESCC.

Radiomics approach for preoperative identification of stages I-II and III-IV of esophageal cancer.

ObjectiveTo predict preoperative staging using a radiomics approach based on computed tomography (CT) images of patients with esophageal squamous cell carcinoma (ESCC).MethodsThis retrospective study included 154 patients (primary cohort: n=114; validation cohort: n=40) with pathologically confirmed ESCC. All patients underwent a preoperative CT scan from the neck to abdomen. High throughput and quantitative radiomics features were extracted from the CT images for each patient. A radiomics signature was constructed using the least absolute shrinkage and selection operator (Lasso). Associations between radiomics signature, tumor volume and ESCC staging were explored. Diagnostic performance of radiomics approach and tumor volume for discriminating between stages I−II and III−IV was evaluated and compared using the receiver operating characteristics (ROC) curves and net reclassification improvement (NRI).ResultsA total of 9,790 radiomics features were extracted. Ten features were selected to build a radiomics signature after feature dimension reduction. The radiomics signature was significantly associated with ESCC staging (P<0.001), and yielded a better performance for discrimination of early and advanced stage ESCC compared to tumor volume in both the primary [area under the receiver operating characteristic curve (AUC): 0.795vs. 0.694, P=0.003; NRI=0.424)] and validation cohorts (AUC: 0.762 vs. 0.624, P=0.035; NRI=0.834). ConclusionsThe quantitative approach has the potential to identify stage I−II and III−IV ESCC before treatment.

Cell-free plasma hypermethylated CASZ1, CDH13 and ING2 are promising biomarkers of esophageal cancer.

Identifying sensitive and specific biomarkers for early detection of cancer is immensely imperative for early diagnosis and treatment and better clinical outcome of cancer patients. This study aimed to construct a specific DNA methylation pattern of cancer suppressor genes and explore the feasibility of applying cell-free DNA based methylation as a biomarker for early diagnosis of esophageal squamous cell carcinoma (ESCC). We recruited early stage ESCC patients from Yangzhong County, China. The Illumina Infinium 450K Methylation BeadChip was used to construct a genome-wide DNA methylation profile. Then, differentiated genes were selected for the validation study using the Sequenom MassARRAY platform. The frequency of methylation was compared between cancer tissues, matched cell-free DNAs and normal controls. The specific methylation profiles were constructed, and the sensitivity and specificity were calculated. Seven CG sites in three genes CASZ1, CDH13 and ING2 were significantly hypermethylated in ESCC as compared with normal controls. A significant correlation was found between the methylation of DNA extracted from cancer tissues and matched plasma cell-free DNA, either for individual CG site or for cumulative methylation analysis. The sensitivity and specificity reached 100% at an appropriate cut-point using these specific methylation biomarkers. This study revealed that aberrant DNA methylation is a promising biomarker for molecular diagnosis of esophageal cancer. Hypermethylation of CASZ1, CDH13 and ING2 detected in plasma cell-free DNA can be applied as a potential noninvasive biomarker for diagnosis of esophageal cancer.

Rho/ROCK Pathway Regulates Migration and Invasion of Esophageal Squamous Cell Carcinoma by Regulating Caveolin-1.

BackgroundEsophageal squamous cell carcinoma (ESCC) is a common cancer with poor prognosis. Caveolin-1 (Cav1) and Rho/ROCK pathway play important roles in tumor metastasis, separately. However, less research was focused on the relationship between Cav1 and Rho/ROCK in ECSS metastasis. Therefore, we investigated the relationship between Cav1 and Rho/ROCK pathway in ESCC metastasis.Material/MethodsCav1 and phosphorylated Cav1 (PY14Cav1) were examined in ESCC and in adjacent and non-tumorous tissues from ESCC patients by immunohistochemistry (IHC). Small interfering RNA (siRNA) targeting Cav1 or Rho/ROCK inhibitor was used to treat EC109, Eca109, TE1, and TE13 cells. Western blotting (WB) was used to detect Cav1 and PY14Cav1 expression. The wound healing scratch test and transwell assays were used to assess migration and invasion.ResultsCav1 and PY14Cav1 were gradually expressed at higher levels in ECSS than in adjacent and non-tumor tissues as ESCC stage and lymphatic metastasis increased, and this difference was significant (P<0.05). Cav1 was expressed at higher levels in TE1 and TE13 than in EC109 and Eca109, while PY14Cav1 was enhanced in TE1 and TE13 cells but not in EC109 and Eca109, and the difference was significant (P<0.05). TE1 and TE13 had significantly (P<0.05) stronger motility, migratory, and invasion abilities than EC109 and Eca109 cells. Silencing Cav1 decreased PY14Cav1 expression in TE1 and TE13 cells, as well as suppressing the migration and invasion of all ECSS cells, and these differences were significant (P<0.05). Suppressing the Rho/ROCK pathway obviously inhibited Cav1 and PY14Cav1 expressions, as well as significantly (P<0.05) decreasing migration and invasion of ESCC cells.ConclusionsCav1 and PY14Cav1 were positively correlated with ESCC lymphatic metastasis and cancer stages. Rho/ROCK pathway activation promoted ESCC metastasis by regulating Cav1.

Expression of CCR6 in esophageal squamous cell carcinoma and its effects on epithelial-to-mesenchymal transition.

Esophageal squamous cell carcinoma (ESCC) is the most common esophageal cancer associated with poor prognosis. We detected the expression of C-C motif chemokine receptor 6 (CCR6) and epithelial-to-mesenchymal transition (EMT) markers in esophageal tissues/cells, and evaluated the effects of CCR6 on ESCC cells proliferation, migration and invasion in response to C-C motif chemokine ligand 20 (CCL20) treatment. Our data showed CCR6 was highly expressed in ESCC cell lines (ECA-109 and TE-1), whereas kept in a low expression in normal cell lines HEEC (P < 0.001). CCL20 stimulus induced a significant decrease in the proliferation ability of ESCC (P < 0.05). The healing speed of CCL20 group was significantly higher than control in ECA-109 (P < 0.01), whereas significantly lower in αCCR6+CCL20 group than CCL20 group (P < 0.05).The number of cells permeabling through the polycarbonate membrane in CCL20 group was higher than control (P < 0.01). The cell number in αCCR6+CCL20 group was significantly reduced compared to CCL20 group in ECA-109 (P < 0.05). Moreover, after CCL20 stimulated in ECA-109, both mRNA and protein level of E-cadherin significantly decreased compared to control, while Vimentin was significantly higher. In αCCR6+CCL20 group, mRNA and protein level of E-cadherin significantly increased compared to CCL20 group, while Vimentin was much lower than CCL20 group. There was no significant difference in TE-1. In summary, high expression of CCR6 existed in the lymph node metastasis and TNM stage of ESCC. CCR6 play an important role in the regulation of tumor cell proliferation, invasion and migration. CCR6 may participate in regulating the occurrence of EMT in ESCC.

Longitudinal study of esophageal mucosal damage after esophagectomy and gastric interposition: relationship between reflux-related mucosal injury and Notch signaling.

Esophagectomy with gastric interposition could serve as a good human reflux model to study the molecular pathogenesis of esophageal mucosal damage induced by gastroesophageal reflux. This study was to investigate the role of Notch signaling in reflux injury of esophageal mucosa. Patients undergoing Ivor-Lewis esophagectomy for early stage esophageal squamous cell carcinoma were included. Follow-ups were scheduled at 6, 18, 36 and 48 months postoperatively, including reflux symptom assessment, endoscopic and histological evaluation of esophageal mucosal damage. The expressions of Notch1 and its downstream target gene Hes1 were evaluated by real-time polymerase chain reaction (PCR) and immunohistochemistry (IHC). Forty-four out of 48 patients completed four follow-ups. Injuries of esophageal remnant confirmed by endoscopical and histological examinations were both more often with a longer postoperative period (P<0.05). The mRNA expression levels of Notch1 and Hes1 were decreased in a time-dependent manner after operation (P<0.001). Notch1 and Hes1 mRNA levels were significantly higher in normal squamous mucosa than in esophagitis, and higher in esophagitis than in metaplasia (P<0.05). Immunohistochemical study also demonstrated a similar protein expression pattern. Samples with endoscopic evidence of mucosal damage exhibited lower expression of Notch1 mRNA levels as compared to biopsies without visualized damage (P=0.035). This is the first longitudinal study on Notch signaling in human esophagectomy model, our preliminary findings suggest decreased Notch signaling might be involved in the development of mucosa damage caused by gastroesophageal reflux.

Serum anti-TOPO48 autoantibody as a biomarker for early diagnosis and prognosis in patients with esophageal squamous cell carcinoma

We previously reported a novel tumor associated antigen (TTA) with molecular weight around 48kDa that is a fragment derived from human DNA-topoiomerase I (TOP1). We termed the novel TAA as TOPO48 and termed autoantibody against the TAA as anti-TOPO48 autoantibody. The aim of this study is to further investigate the clinical applications of the autoantibody in patients with esophageal squamous cell carcinomas (ESCC). Serum levels of the anti-TOPO48 autoantibody in 112 ESCC patients, 112 age- and gender-matched healthy controls and 75 patients with esophageal benign tumors were determined by using a specific anti-TOPO48 autoantibody ELISA. Then, we statistically evaluated its clinical significance. We found that serum anti-TOPO48 autoantibody levels in ESCC patients were significantly higher than that in healthy controls and benign tumor patients (P=0.001). The percentage of sera with a positive level of anti-TOPO48 autoantibody in early stages was significantly higher than that in advanced stages of the cancer patients when the maximum level of healthy control sera was taken as a cut-off value (P=0.001). The area under ROC curve was 0.863 (95% CI=0.797-0.928) for healthy controls vs. early stage ESCC. In addition, patients with positive anti-TOPO48 autoantibody had significantly higher survival rate and longer survival time than that with negative anti-TOPO48 autoantibody in cancer patients (P=0.038, 0.025 and 0.047 for all stages, early stage and advanced stage, respectively). Our results suggest that anti-TOPO48 autoantibody may be a potentially useful biomarker for early diagnosis and prognosis of ESCC.

Texture analysis of CT imaging for assessment of esophageal squamous cancer aggressiveness.

To explore the role of texture analysis of computed tomography (CT) images in preoperative assessment of esophageal squamous cell carcinoma (ESCC) aggressiveness. Seventy-three patients with pathologically confirmed ESCC underwent unenhanced and contrast enhanced CT imaging preoperatively. Texture analysis was performed on unenhanced and contrast enhanced CT images, respectively. Six CT texture parameters were obtained. One-way analysis of variance or independent-samples t-test (normality), independent-samples Kruskal-Wallis test or Mann-Whitney U test (non-normality), binary Logistic regression analysis (multivariable), Spearman correlation test, receiver operating characteristic (ROC) curve analysis and intraclass correlation coefficient (ICC) were used for statistical analyses. Kurtosis was an independent predictor for T stages (T1-2 vs. T3-4) as well as overall stages (I-II vs. III-IV) based on unenhanced CT images, while entropy was an independent predictor for T stages (T1-2 vs. T3-4), lymph node metastasis (N- vs. N+) and overall stages (I/II vs. III/IV). Skew and kurtosis based on unenhanced CT images showed significant differences among N stages (N0, N1, N2 and N3) as well as 90th percentile based on contrast enhanced CT images. In correlation with T stage of ESCC, kurtosis and entropy significantly correlated with T stage both on unenhanced and contrast enhanced CT images. Reversely, entropy and 90th percentile based on contrast enhanced CT images showed significant correlations with N stage (r: 0.526, 0.265; both P<0.05), as well as overall stage (r: 0.562, 0.315; both P<0.05). For identifying ESCC with different T stages (T1-2 vs. T3-4), lymph node metastasis (N- vs. N+) and overall stages (I/II vs. III/IV), entropy based on contrast enhanced CT images, showed good performance with area under ROC curve area under curve (AUC) of 0.637, 0.815 and 0.778, respectively. Texture analysis of CT images held great potential in differentiating different T, N and overall stages of ESCC preoperatively, while failed to assess the differentiation degrees.

The management of localized esophageal squamous cell carcinoma: Western approach.

Esophageal cancer remains a significant cause of cancer-related mortality worldwide and carries a poor prognosis both in the locally advanced and metastatic setting. Esophageal squamous cell carcinoma (ESCC) accounts for the majority of esophageal cancers globally, while esophageal adenocarcinoma (EAC) now predominates in the Western world. Historically, clinical trials have not separated the two histologic subtypes, with similar regimens being used for both. This, however, is changing with the recent publication by the Cancer Genome Atlas Research Group confirming that comprehensive analysis reveals ESCC demonstrates frequent genomic amplifications of CCND1 and SOX2 and/or TP63 and biologically are more like head and neck squamous cell carcinomas, whereas EACs strongly resemble the chromosomally unstable variant of gastric cancer. With a lack of viable targets in squamous cell carcinoma, immunotherapeutic approaches are now being investigated in both early stage and metastatic ESCC with promising results being reported in the preliminary data published to date.

High amplification of FGFR1 gene is a delayed poor prognostic factor in early stage ESCC patients.

Amplification of the fibroblast growth factor receptor 1 (FGFR1) is believed to predict response to FGFR inhibitors. The aim of this study was to investigate the frequency and the prognostic impact of FGFR1 amplification in patients with resected esophageal squamous cell carcinoma (ESCC) by using fluorescent in situ hybridization. Microarrayed paraffin embedded blocks were constructed, and the cohort of tissues came from 506 patients with ESCC. FGFR1 high amplification (FGFR1high) was defined by an FGFR1/centromere 8 ratio of ≥ 2.0, or average number of FGFR1 signals/tumor cell nucleus ≥ 6.0, or percentage of tumor cells containing ≥ 15 FGFR1 signals, or large cluster in ≥ 10% of cancer cells. FGFR1 low amplification was defined by ≥ 5 FGFR1 signals in ≥ 50% of cancer cells. Kaplan-Meier curves with log-rank tests and Cox proportional hazards model were used to analyze patients’ survival. Among 506 patients, high amplification, low amplification, and disomy were detected in 8.7%, 3.6% and 87.7%, respectively. In general, the FGFR1high group trended towards worse disease-free survival (DFS) and overall survival (OS) compared to the FGFR1 low amplification/disomy (FGFR1low/disomy) group (DFS, P=0.108; OS, P=0.112), but this trend was amplified for patients with DFS ≥ 30 months (DFS, P=0.009; OS, P=0.007). Furthermore, when patients were stratified into stage I-II and stage III-IV, the FGFR1high group directly presented with adverse DFS and OS than the FGFR1low/disomy group in stage I-II patients (DFS, P=0.019; OS, P=0.034), especially with DFS ≥ 30 months (DFS, P=0.002; OS, P=0.001). However, for patients in stage III-IV, FGFR1high had no effect on prognosis regardless of DFS time. FGFR1high occurs in a minority of ESCC, and it predicts delayed poor prognosis in stage I and II ESCC patients.

Thymidine phosphorylase and angiogenesis in early stage esophageal squamous cell carcinoma.

The relationship between thymidine phosphorylase (TP) and angiogenesis at the early stage of esophageal squamous cell carcinoma has been unclear. Using 14 samples of normal squamous epithelium, 11 samples of low-grade intraepithelial neoplasia, and 64 samples of superficial esophageal cancer, microvessel density (MVD) was estimated using immunostaining for CD34 and CD105. TP expression was also evaluated in both cancer cells and stromal monocytic cells (SMCs). We then investigated the correlation between MVD and TP expression in both cancer cells and SMCs. On the basis of the above parameters, MVD was significantly higher in cancerous lesions than in normal squamous epithelium. In terms of CD34 and CD105 expression, MVD showed a gradual increase from normal squamous epithelium, to low-grade intraepithelial neoplasia, and then to M1 and M2 cancer, and M3 or deeper cancer. M1 and M2 cancer showed overexpression of TP in both cancer cells and SMCs. There was no significant correlation between TP expression in cancer cells and MVD estimated from CD34 (rS=0.16, P=0.21) or CD105 (rS=0.05, P=0.68) expression. Significant correlations were found between TP expression in SMCs and CD34-related (rS=0.46, P<0.001) and CD105-related (rS=0.34, P<0.01) MVD. In M3 or deeper cancers, there were no significant correlations between TP expression in cancer cells or SMCs and venous invasion, lymphatic invasion, and lymph node metastasis. TP expression is activated in both cancer cells and stromal monocytic cells at the very early stage of ESCC progression. TP expression in SMCs, rather than in cancer cells, is significantly correlated with angiogenesis.