Early Stage Esophageal Squamous Cell Research Articles

Tumor-associated macrophages and angiogenesis in early stage esophageal squamous cell carcinoma

The association between angiogenesis and tumor-associated macrophages (TAMs) is unclear. Mononuclear cell infiltration was reported to induce angiogenesis in early stage esophageal squamous cell carcinoma (ESCC). The study materials included 14 samples of normal squamous epithelium, 11 samples of low-grade intraepithelial neoplasia, and 64 samples of superficial esophageal cancer (M1 and M2 cancer 27; M3 or deeper cancer 37). We assessed microvessel density (MVD) using CD34 and CD105 immunostaining and monocyte count (MC) using CD68 and CD163 immunostaining in relation to the histological type or grade of mononuclear cell infiltration, as well as the correlation between MVD and MC. MVD and MC increased in accordance with histological type, and the differences were significant (P < 0.001). MVD and MC were significantly higher in M1 and M2 lesions than in normal squamous epithelium (P < 0.05). MVD (CD34 and CD105) and MC (CD68 and CD163) were significantly correlated with the degree of mononuclear cell infiltration (P < 0.001), and there was a strong correlation between MC assessed using CD68 and MC assessed using CD163 (rS = 0.93, P < 0.001). The CD163/CD68 ratio did not differ significantly according to histological type. There was a significant correlation between MVD assessed using CD105 and MC assessed using CD68 (rS = 0.69, P < 0.001) and CD163 (rS = 0.67, P < 0.001). MVD assessed using CD34 was also significantly correlated with MC assessed using both CD68 (rS = 0.59, P < 0.001) and CD163 (rS = 0.57, P < 0.001). The number of TAMs is significantly associated with the development of neovasculature in the early stage of ESCC progression.

Combination of platelet count and mean platelet volume (COP-MPV) predicts postoperative prognosis in both resectable early and advanced stage esophageal squamous cell cancer patients.

The aim of this study is to search the most powerful prognostic factor from routine blood test for esophageal squamous cell cancer (ESCC) patients. Multiple laboratory tests were evaluated including those reflecting red blood cell parameters (hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and red blood cell distribution width (RDW)), platelet morphological parameters (mean platelet volume (MPV) and platelet count (PLT)), blood coagulation status (D-dimer), and tumor biomarker (CA19-9). Known inflammatory indices (NLR and PLR) were also calculated. A total of 468 patients who were diagnosed with ESCC between December 2005 and December 2008 were retrospectively analyzed in this study. By utilizing univariate and multivariate Cox proportional hazard analyses, we found that PLT and MPV were significantly associated with overall survival (OS) and disease-free survival (DFS) of ESCC patients, with optimal cutoff values of 212 and 10.6, respectively. Moreover, the combination of the preoperative PLT and MPV (COP-MPV) was calculated as follows: patients with both PLT (≥212 × 109 L−1) and MPV (≥10.6 fL) elevation were assigned a score of 2, and patients with one or neither were assigned a score of 1 and 0. The COP-MPV was an independent prognostic factor for OS (hazard ratio (HR) 0.378, 95 % confidence interval (CI) 0.241 to 0.593, P < 0.001, 0/2) and DFS (HR 0.341, 95 % CI 0.218 to 0.534, P < 0.001, 0/2) in multivariate analyses. In subgroup analyses for early (stages I and II) and locally (stage III) advanced stage patients, COP-MPV was found significantly associated with OS and DFS in each group (P = 0.025 and P = 0.018 for OS and P = 0.029 and P = 0.002 for DFS). In conclusion, we considered that COP-MPV is a promising predictor for postoperative survival in ESCC patients.

Coexpression of COX-2 and iNOS in Angiogenesis of Superficial Esophageal Squamous Cell Carcinoma.

Using immunohistochemical staining, the present study was conducted to examine whether cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) affect angiogenesis in early-stage esophageal squamous cell carcinoma (ESCC). We also analyzed the correlation between these two factors. Cyclooxygenase 2, iNOS, and angiogenesis in early-stage ESCC are unclear. Using 10 samples of normal squamous epithelium, 7 samples of low-grade intraepithelial neoplasia (LGIN), and 45 samples of superficial esophageal cancer, we observed the expression of COX-2 and iNOS. We then investigated the COX-2 and iNOS immunoreactivity scores and the correlation between COX-2 or iNOS scores and microvessel density (MVD) using CD34 or CD105. The intensity of COX-2 or iNOS expression differed significantly according to histological type (P < 0.001). The scores of COX-2 and iNOS were lowest for normal squamous epithelium, followed in ascending order by LGIN, carcinoma in situ and tumor invading the lamina propria mucosae (M1-M2 cancer); and tumor invading the muscularis mucosa (M3) or deeper cancer. The differences were significant (P < 0.001). Cancers classified M1-M2 (P < 0.01 and P < 0.05, respectively); M3; or deeper cancer (P < 0.01) had significantly higher COX-2 and iNOS scores than normal squamous epithelium. There was a significant correlation between COX-2 and iNOS scores (P < 0.001, rs = 0.51). Correlations between COX-2 score and CD34-positive MVD or CD105-positive MVD were significant (rs = 0.53, P < 0.001; rs = 0.62, P < 0.001, respectively). Inducible nitric oxide synthase score was also significantly correlated with CD34 MVD and CD105 MVD (rs = 0.45, P < 0.001; rs = 0.60, P < 0.001, respectively). Chemoprevention of COX-2 or iNOS activity may blunt the development of ESCC from precancerous lesions.

Expression profiling of metastasis-related microRNAs in early esophageal squamous cell carcinoma

To study the difference of microRNA (miRNA) expression between two groups of early stage (pT1N0) esophageal squamous cell carcinoma (ESCC) patients who had different outcome and the prognostic significance of different miRNA in metastatic of early ESCC, and to identify useful prognostic markers in the selection of appropriate treatment for early ESCC patients. TaqMan human miRNA arrays and bioinformatics were used to detect and analyze the expression profiles of miRNAs in the two groups, and RT-PCR was used to verify the differences in miRNA expression. The miRNA arrays revealed a total of 41 markedly changed miRNAs in the survival group compared with the death group. Bioinformatics analysis, prediction and significant function analyses of targeted genes and pathway analysis identified that miR-27a, miR-143 and miR-886-5p levels were increased or decreased by seven-folds or more. The enriched target genes were GRB2, SOS1, MAPK1, EGFR, CBL, SPRY2, RPS6KA5, IGF1R, NGFR, MAPK14 and CREB1. These genes were significantly related to the following signaling pathways, i.e.Sprouty regulation of tyrosine kinase signals pathway, Erk1/Erk2 Mapk signaling pathway and transcription factor CREB and its extracellular signals. miR-27a, miR-886-5p, and miR-143 may be potential prognostic markers of metastasis for early ESCC. The detection of these miRNAs plays a directive role for the treatment options of early ESCC. The regulation of targeted genes and mechanism remain to be further studied.

Downregulation of 14-3-3σ correlates with multistage carcinogenesis and poor prognosis of esophageal squamous cell carcinoma.

AimsThe asymptomatic nature of early-stage esophageal squamous cell carcinoma (ESCC) results in late presentation and consequent dismal prognosis This study characterized 14-3-3σ protein expression in the multi-stage development of ESCC and determined its correlation with clinical features and prognosis.Materials and MethodsWestern blot was used to examine 14-3-3σ protein expression in normal esophageal epithelium (NEE), low grade intraepithelial neoplasia (LGIN), high grade intraepithelial neoplasia (HGIN), ESCC of TNM I to IV stage and various esophageal epithelial cell lines with different biological behavior. Immunohistochemistry was used to estimate 14-3-3σ protein in 110 biopsy samples of NEE, LGIN or HGIN and in 168 ESCC samples all of whom had follow-up data. Support vector machine (SVM) was used to develop a classifier for prognosis.Results14-3-3σ decreased progressively from NEE to LGIN, to HGIN, and to ESCC. Chemoresistant sub-lines of EC9706/PTX and EC9706/CDDP showed high expression of 14-3-3σ protein compared with non-chemoresistant ESCC cell lines and immortalized NEC. Furthermore, the downregulation of 14-3-3σ correlated significantly with histological grade (P = 0.000) and worse prognosis (P = 0.004). Multivariate Cox regression analysis indicated that 14-3-3σ protein (P = 0.016) and T stage (P = 0.000) were independent prognostic factors for ESCC. The SVM ESCC classifier comprising sex, age, T stage, histological grade, lymph node metastasis, clinical stage and 14-3-3σ, distinguished significantly lower- and higher-risk ESCC patients (91.67% vs. 3.62%, P = 0.000).ConclusionsDownregulation of 14-3-3σ arises early in the development of ESCC and predicts poor survival, suggesting that 14-3-3σ may be a biomarker for early detection of high-risk subjects and diagnosis of ESCC. Our seven-feature SVM classifier for ESCC prognosis may help to inform clinical decisions and tailor individual therapy.

351 Long-Term Outcomes of Endoscopic Resection for Early-Stage Esophageal Squamous Cell Neoplasms: a Multicenter Prospective Study

The efficacy and limitations of endoscopic resection (ER) for esophageal superficial squamous cell neoplasms have been reported; however, most previous studies were retrospective and/or single center studies. There has been no prospective multicenter study with a large sample size.

Preoperative elevation of serum C-reactive protein as an indicator of poor prognosis for early-stage esophageal squamous cell carcinoma

Preoperative elevation of serum C-reactive protein (CRP) is reportedly associated with poor prognosis in several types of cancer. This study investigated the role of serum CRP as a prognostic factor in early-stage esophageal squamous cell carcinoma (ESCC). The preoperative serum CRP levels were measured in 156 newly diagnosed pT1–2N0M0 patients using an enzyme-linked immunosorbent assay. Correlations between serum CRP levels and other clinical parameters were analyzed. Multivariate analyses were performed to find prognostic markers using Cox's proportional hazards model. CRP concentrations were within the normal range in 117 (75%) individuals, but were elevated in 39 (25%) patients. Serum CRP levels were significantly correlated with the tumor length (p = 0.032), depth (T classification, p = 0.0157), or histologic grade (p = 0.034). The overall 5-year survival rates were 76.3% and 50.2% in the low- and high-CRP groups, respectively (p = 0.005). By multivariate analyses, the elevated serum CRP level was found to be an independent prognostic factor for poor survival (hazard ratio = 2.131; p = 0.007), regardless of tumor classification or other prognostic factors. In conclusion, preoperative, high serum CRP is an independent determinant of poor prognosis in early-stage ESCC.

Early-Stage Esophageal Squamous Cell Carcinoma Treated with Californium-252 Neutron Brachytherapy: Clinical Report on 16 Cases

Californium-252 (²⁵²Cf) neutron brachytherapy is a form of high linear energy transfer radiotherapy, which has proven effective when used in combination with external beam radiotherapy to treat intracavitary cancers of the cervix, colon/rectum and esophagus. No study has been reported for treatment of intracavitary cancers with neutron brachytherapy alone. The aim of the study was to observe and analyze the long-term curative effects and complications for early stage thoracic esophageal cancer patients treated with neutron brachytherapy alone. From December 2001 to August 2006, 16 patients of early stage squamous cell carcinoma underwent neutron brachytherapy. The total radiation dose to the reference point was 20-28 Gy-eq in 5 to 7 fractions with 4 Gy-eq/fraction. The 1-, 3-, and 5-year follow-up rates were 100%. The 2-, 3-, 4-, and 5-year survival rates were 100%, 87.5%, 87.5%, and 75%, respectively. The early complication rates for grades 1 and 2 radiation esophagitis were 75% and 25%, respectively. The late complication rates for grades 0 and 1 (according to the RTOG/EORTC standard) were 87.5% and 12.5%, respectively. Barium esophagography after treatments confirmed that the complete response rate was 100%. Fourteen patients were confirmed by endoscopy to have either normal mucosa or inflammation change. Neutron brachytherapy alone was an effective and safe treatment for early stage esophageal squamous cell cancer.

Analysis of the tumor length and other prognosis factors in pT1-2 node-negative esophageal squamous cell carcinoma in a Chinese population.

BackgroundTumor length is an important prognostic factor for many carcinomas, but its role in esophageal cancer remained undetermined. The aim of this study was to investigate the effect of tumor length on survival for patients with confined tumors (grade pT1-2) without lymph-node metastases in esophageal squamous cell carcinoma.MethodsWe enrolled 201 patients with esophageal squamous cell carcinoma (SCC) who had undergone surgical resection and been confirmed as pT1-2N0M0. The relationship of tumor length with overall survival was assessed and compared with other factors detailed in the American Joint Committee on Cancer (AJCC) tumor, node, metastasis (TNM) staging system published in 2009.ResultsThe overall survival (OS) rates at 1, 3, and 5 years were 93.0%, 83.7%, and 69.2%, respectively. The tumor length adversely affected OS, with the 5-year rate being 93.5%, 82.0%, 68.6%, 67.9%, 55.3% and 41.1%, respectively for tumor lengths of less than 10 mm, 10 to 20 mm, 20 to 30 mm, 30 to 40 mm, 40 to 50 mm, and greater than 50 mm (P< 0.001). Multivariate analyses showed that the pathologic T classification and grade of tumor was significantly associated with OS. Tumor length of 30 mm or more remained an independent prognostic factor (P = 0.04), as did the other current TNM factors.ConclusionTumor length appears to affect the OS of patients with early-stage esophageal squamous cell carcinoma. It may provide additional prognostic information for the current TNM staging system.

Optical Imaging of Periostin Enables Early Endoscopic Detection and Characterization of Esophageal Cancer in Mice

Imaging strategies that detect early stage esophageal squamous cell carcinoma (ESCC) could improve clinical outcomes, when combined with endoscopic approaches. Periostin is an integrin-binding protein that is important in the tumor microenvironment. We created a fluorescent-labeled antibody that recognizes periostin and binds specifically to ESCC xenograft tumors in mice. In L2-cre;p120ctnLoxP/LoxP mice, which develop squamous cell cancers that resemble human ESCC, we visualized the probe in preneoplastic and neoplastic esophageal lesions using near-infrared fluorescent imaging with upper-gastrointestinal endoscopy. Periostin might be a biomarker of the esophageal tumor microenvironment that can be used to detect preneoplastic lesions.

Early-stage primary malignant melanoma of the esophagus detected simultaneously with esophageal squamous cell carcinoma

Primary malignant melanoma of the esophagus is a rare disease. The majority of patients are diagnosed at an advanced stage, and only a few are detected at an early stage. We herein describe a case of early-stage primary malignant melanoma of the esophagus that was detected simultaneously with early-stage primary esophageal squamous cell carcinoma. Both tumors were detected during esophagogastroduodenoscopy for heartburn. The malignant melanoma tumor was a nevus-like flat-type lesion in the upper thoracic esophagus, and the squamous cell carcinoma was a slightly depressed lesion in the abdominal esophagus. The tumor was resected by thoracoscopic esophagectomy. Histologically, the invasion of both tumors was limited to the mucosal layer, and no lymph node metastasis was detected. Immunohistochemically, the malignant melanoma cells were strongly positive for HMB-45, melan-A, and S-100 protein. The patient has survived without recurrence for 17 months after the operation.

Indications, techniques, and outcomes of endoscopic submucosal dissection for esophageal squamous cell carcinoma

Endoscopic submucosal dissection (ESD) has gradually gained acceptance as one of the standard treatments for esophageal squamous cell carcinoma in Japan. Principal indications are little likelihood of nodal metastasis and having technical resectability. All lesions with preoperative diagnosis of high-grade intraepithelial neoplasms, including carcinomas in situ (T1a-EP), carcinomas invading the lamina propria (T1a-LPM), and some with preoperative diagnosis of carcinomas invading the muscularis mucosa (T1a-MM) and minute submucosal invasion (T1b-SM1), may become candidates for ESD, considering the substantial risks and benefits to be obtained. ESD as a staging measure to obtain histological confirmation of invasion depth and lymphovascular infiltration may be acceptable because preoperative prediction is difficult in some cases. In terms of technique, advantages of ESD in comparison with other endoscopic treatment are controllability in size and shape, and that even large and fibrotic neoplasms can be resected. Disadvantages may be longer procedure time, heavier bleeding, and greater possibility of perforation. However, with refinement of the technique, new devices, and the learning curve, acceptable technical safety has been achieved. Our experience with 104 consecutive cases shows favorable outcomes, although further innovation may be necessary. We are convinced that ESD will become a standard treatment for early-stage esophageal squamous cell carcinoma not only in Japan but also all over the world in the near future.

Analyzing the profile of chromosomal imbalances in esophageal atypical hyperplasia and early stage esophageal squamous cell carcinoma by 250K Snp Array

To detect DNA copy number abnormality and LOH and explore the profile of chromosomal imbalances in esophageal atypical hyperplasia and early stage esophageal squamous cell carcinoma using SNP array. The DNA copy number abnormality and LOH in pathological change esophageal tissue and matched normal esophageal tissue of one case of primary esophageal high atypical hyperplasia and four cases of primary early stage esophageal squamous cell carcinoma were detected by using Affymetrix GeneChip Human Mapping 250K Nsp Array. Amplification was found in a few DNA fragment and no deletion or LOH was found in esophageal high atypical hyperplasia. In early stage esophageal squamous cell carcinoma, DNA amplification occurred in 1p, 1q, 2p, 2q, 3q, 4q, 5p, 6p, 6q, 7q, 8q, 11p, 11q, 12p, 12q, 14q, 17q, 18p, 19q, 20q, 22q and X chromosome. DNA deletion occurred in 1p, 2q, 3p, 3q, 4p, 4q, 8p, 9p, 9q, 10q, 11p, 13q, 16p, 18q, 19p, 19q and 22q chromosome. LOH occurred in 3q and 9q chromosome. The amplification in 1p, 19q, 4q, 11p chromosome and the deletion in 3q, 11p, 2q, 16p chromosome were rarely reported. DNA abnormality was rare in esophageal high atypical hyperplasia. The obvious amplification and deletion in DNA have been found in early stage esophageal squamous cell carcinoma, but LOH was rarely found. 250K SNP array could effectively detect DNA copy number change and LOH in whole-wide genes in esophageal atypical hyperplasia and early stage esophageal squamous cell carcinoma with high distinguishability and precise location. The results would provide important academic information for detection and location of related genes in early stage esophageal squamous cell carcinoma.

CpG island hypermethylation of E‐cadherin (CDH1) and integrin α4 is associated with recurrence of early stage esophageal squamous cell carcinoma

The prognosis of esophageal squamous cell carcinoma (ESCC) patients remains very poor, which is partially due to a high rate of recurrence. This study was aimed at identifying a recurrence-associated epigenetic prognostic marker in patients with ESCC. We retrospectively analyzed the CpG island hypermethylation of the p16, Wif-1, sFRP1, integrin alpha4, CDH1, DAP kinase and RARbeta2 genes in 251 ESCCs. The methylation status was determined by methylation-specific PCR. Hypermethylation was detected in 52% for p16, 25% for RARbeta2, 43% for CDH1, 21% for integrin alpha4, 57% for sFRP1, 38% for DAP kinase and 35% for Wif-1. Recurrence was observed in 131 (52%) of the 251 cases. For stage I cancers, CDH1 methylation was associated with a high risk of recurrence (OR = 5.26, 95% CI = 1.48-18.67; p = 0.01) and a poor recurrence-free survival after surgery (HR = 3.13, 95% CI = 1.21-8.09; p = 0.02). The hazard of failure after recurrence was about 13.17 (95% CI = 2.46-70.41; p = 0.003) times higher in patients with Wif-1 methylation than in those without. For stage II cancers, integrin alpha4 methylation was associated with an increased risk of recurrence (OR = 3.03, 95% CI = 1.09-8.37; p = 0.03) and a poor recurrence-free survival (HR = 2.12, 95% CI = 1.13-3.98; p = 0.03). In conclusion, the present study suggests that hypermethylation of CDH1 and integrin alpha4 genes may be used as recurrence-associated prognostic indicators in stage I and stage II ESCCs, respectively.

Aberrant methylation of Fragile Histidine Triad gene is associated with poor prognosis in early stage esophageal squamous cell carcinoma

The aim of this study was to understand the clinicopathological and prognostic significance of promoter methylation of Fragile Histidine Triad ( FHIT) gene in esophageal cancer. FHIT methylation in 257 primary esophageal squamous cell carcinomas was retrospectively analyzed by methylation-specific polymerase chain reaction. Aberrant methylation of FHIT was found in 85 (33%) of 257 esophageal cancer patients. The FHIT methylation was found to be significantly associated with exposure to tobacco smoke ( P = 0.007) and with a poor prognosis in cases of stage 1–2 cancer irrespective of recurrence. The hazard of failure after esophagectomy for stage 1–2 cancers with FHIT methylation was about 5.81 (95% CI = 1.15–14.07; P = 0.009) times higher than in those without. Recurrence occurred in 116 (45%) of the 257 patients studied. The survival after recurrence in stage 1–2 cancers was also poorer for patients with FHIT methylation than in those without (HR = 2.31; 95% CI = 1.18–7.92; P = 0.03). In conclusion, aberrant methylation of the FHIT promoter was found to be significantly associated with exposure to tobacco smoke and with a poor prognosis for stage 1–2 cases, but not with recurrence rate. Our study suggests that FHIT promoter methylation may be an independent prognostic biomarker in early stage esophageal squamous cell carcinoma.