Esophageal Squamous Cell Dysplasia Research Articles

Corrigendum to ‘‘EsophaCap Sponge Cytology Screening for Esophageal Squamous Cell Dysplasia and Carcinoma is Feasible in a High-Risk Area in Western India’’

Corrigendum to ‘‘EsophaCap Sponge Cytology Screening for Esophageal Squamous Cell Dysplasia and Carcinoma is Feasible in a High-Risk Area in Western India’’

EsophaCap Sponge Cytology Screening for Esophageal Squamous Cell Dysplasia and Carcinoma is Feasible in a High-Risk Area in Western India

Objective: Conventional endoscopy remains the gold standard for detecting esophageal squamous cell carcinoma (ESCC) despite its high cost and need for expertise. In resource-poor regions, a less labor-intensive yet accurate screening tool is needed. The purpose of this study is to assess feasibility of establishing a non-endoscopic screening program in rural India and to test the diagnostic accuracy of the EsophaCap swallowable sponge in detecting ESCC in a high-risk patient population. Methods: A prospective cohort study was conducted between 2017 and 2019, in which subjects with risk factors for ESCC (tobacco smoking or chewing, betel nut/leaf, alcohol and hot beverage consumption) were approached during upper endoscopy visits at a clinic in Western India. After obtaining EsophaCap sponge cytology samples, random endoscopic biopsies were obtained at 20 and 30 cm from the incisors. Histologic diagnoses were confirmed and select biopsy samples were sequenced for genomic aberrations. Visual Analog Scale (VAS) scores were used to assess patient experience of sponge swallowing (range 1-5, “very comfortable” to “very uncomfortable”). Results: In our cohort of 178 patients, 157 (88%) were males. Mean age was 52 ± 12 years. Sixty-eight (38%) patients were current cigarette or bidi smokers; 132 (74%) patients were daily tobacco chewers, and 83 (47%) patients chewed betel nuts/leaves on a daily basis. Forty-six (26%) patients were daily alcohol users and 151 (85%) patients drank ≥3 hot beverages per day. The median number of risk factors per patient was 3. The first-time swallow rate of the encapsulated sponge was successful in 190/200 (95%) patients. Median VAS score was 2 (“comfortable”). EsophaCap cytology revealed 6 (3%) patients with atypical squamous cells of unknown significance (ASCUS) and 3 (1%) patients with dysplasia. Based on the endoscopic biopsies, 6 (3%) patients had ESCC, 4 (2%) patients had lesions with squamous dysplasia, and 63 (35%) patients had esophageal leukoplakia. Four patients classified as ASCUS pathology via EsophaCap were normal, benign, or leukoplakia via endoscopy. EsophaCap’s sensitivity and specificity for detecting dysplasia or ESCC by histology was 30% and 97%, respectively. Conclusions: Establishing a non-endoscopic screening program in a high-risk area with language barriers and low medical literacy is very safe and feasible. EsophaCap may help identify ASCUS patients in need of serial endoscopic monitoring. Further studies of combined sponge cytology with immunohistochemistry studies are necessary to improve accuracy of ESCC screening.

Somatic CDKN2A copy number variations are associated with the prognosis of esophageal squamous cell dysplasia.

Somatic copy number variations (SCNVs) in the CDKN2A gene are among the most frequent events in the dysplasia-carcinoma sequence of esophageal squamous cell carcinoma. However, whether CDKN2A SCNVs are useful biomarkers for the risk stratification and management of patients with esophageal squamous cell dysplasia (ESCdys) is unknown. This study aimed to investigate the characteristics and prognostic value of CDKN2A SCNVs in patients with mild or moderate (m/M) ESCdys. This study conducted a prospective multicenter study of 205 patients with a baseline diagnosis of m/M ESCdys in five high-risk regions of China (Ci County, Hebei Province; Yanting, Sichuan Province; Linzhou, Henan Province; Yangzhong, Jiangsu Province; and Feicheng, Shandong Province) from 2005 to 2019. Genomic DNA was extracted from paraffin biopsy samples and paired peripheral white blood cells from patients, and a quantitative polymerase chain reaction assay, P16-Light, was used to detect CDKN2A copy number. The cumulative regression and progression rates of ESCdys were evaluated using competing risk models. A total of 205 patients with baseline m/M ESCdys were enrolled. The proportion of ESCdys regression was significantly lower in the CDKN2A deletion cohort than in the diploid and amplification cohorts (18.8% [13/69] vs. 35.0% [28/80] vs. 51.8% [29/56], P <0.001). In the univariable competing risk analysis, the cumulative regression rate was statistically significantly lower ( P = 0.008), while the cumulative progression rate was higher ( P = 0.017) in ESCdys patients with CDKN2A deletion than in those without CDKN2A deletion. CDKN2A deletion was also an independent predictor of prognosis in ESCdys ( P = 0.004) in the multivariable analysis. The results indicated that CDKN2A SCNVs are associated with the prognosis of ESCdys and may serve as potential biomarkers for risk stratification.

428. CHARACTERIZATION OF CD82/KAI1 IN ESOPHAGEAL ADENOCARCINOMA

Abstract The incidence of esophageal adencocarcinoma (EAC) has increased in last decades. Regardless to improved perioperative chemotherapies and surgical approaches, the prognosis for EAC in advanced stages is poor. CD82 is a tumor suppressor gene, and its downregulation correlates with invasive growth, metastases formation and advanced clinical stages. CD82 might influence the Wnt/ß-catenin signaling pathway. We used an in vitro Barrett’s esophagus cell culture model, represented by esophageal squamous cell epithelium (EPC1, EPC2), metaplasia (CP-A), dysplasia (CP-B) and esophageal adenocarcinoma (OE33, OE19, SKGT4, FLO1). Antisense LNA Oligonucleotides (GapmeRs, Qiagen) were used to achieve a transient knockdown of CD82 in OE33 and OE19 cells. mRNA and proteins were analyzed by q-RT-PCR and western blot analyses, respectively. A fluorescence antibody specific to CD82 was used for flow cytometry analysis. Proliferation assay, colony formation assay and boyden chamber assay were performed after CD82-knockdown to further analyze cell proliferation, cell growth and survival, colony forming and cell migration. CD82 was expressed in all investigated cell lines by flow cytometry with a significant overexpression in OE19 and FLO1 cells. A stable downregulation of CD82 was achieved in OE33 and OE19 cells. Proliferation assays showed a significantly higher proliferation rate at 72 hours after CD82-knockdown in OE33 cells. A significantly higher mRNA expression of vimentin was observed after CD82-knockdown in OE33. Colony formation assay showed a significantly higher colony count after CD82-knockdown in both cell lines. Cell migration was increased after CD82-downregulated in OE33 and OE19 cells. CD82 might influence the expression of mesenchymal marker proteins, cell migration abilities and the process of colony forming and thus enhance the metastatic potential of OE33 and OE19 cells. Low expression of CD82 could alter metastasis and cancer progression but further molecular characterizations are needed to elucidate the impact on carcinogenesis of EAC.

Feasibility of using P16 methylation as a cytologic marker for esophageal squamous cell carcinoma screening: A pilot study.

Early diagnosis and treatment of esophageal squamous cell dysplasia (ESCdys) and esophageal squamous cell carcinoma (ESCC) could significantly reduce the incidence and mortality of ESCC. This pilot study aimed to investigate whether P16/CDKN2A methylation could serve as a cytologic biomarker for early detection of ESCdys and ESCC. Paired esophageal biopsy and cytology specimens (exfoliated cells) were obtained from subjects at different stages of ESCC development. The methylation status of P16 gene in these two specimen types was determined using a 115-bp MethyLight assay. Categorical data were compared by the Chi-square test. Logistic regression was performed to assess adjusted odds ratios of P16 methylation associated with ESCC and ESCdys. Prediction models for identifying individuals at risk of ESCC and high-grade ESCdys (high-grade intraepithelial neoplasia, HGIN) were developed by multivariable logistic regression. Diagnostic performance was evaluated using receiver operating characteristic (ROC) analysis. Internal validation of the prediction models was performed using the 1000-bootstrap resample. A total of 105 subjects with diagnoses ranging from normal mucosa through ESCC were included in this study. An increase in P16 methylation frequency was observed with increasing severity of esophageal lesions (p for trend <0.001). In the adjusted logistic regression models, P16 methylation in cytology specimens was positively associated with ESCC and ESCdys risk, whereas P16 methylation in biopsy specimens was only associated with a higher risk of developing ESCC. The predictive capacity of base model I (AUC, 0.816) for ESCC and HGIN was significantly increased by adding P16 methylation in cytology specimens (model III; AUC, 0.882; p=0.043), but not P16 methylation in biopsy specimens (model II; AUC, 0.850; p=0.225). Bootstrap validation showed optimism-corrected AUC of 0.789 for model I, 0.822 for model II, and 0.854 for model III. P16 methylation as a cytologic marker was associated with the ESCC development and has the potential for application in minimally invasive ESCC screening.

A fractal based approach to evaluate the progression of esophageal squamous cell dysplasia

Abstract Esophageal squamous cell carcinoma (ESCC) is the most prevalent malignancy of the esophagus with a very poor prognosis. Nevertheless, squamous cell dysplasia (ESD) has been identified as the only histological precursors of ESCC. Since, tissue alterations are slight in the early stage of ESD, human diagnosis is subjective. Hence, this work presents a first computer-aided system to differentiate low-grade dysplasia (LGD) from normal esophageal mucosa according to Vienna grading system, which is the most widespread method for histological grading of esophagus tissues. We captured microscopic images of a well-oriented region of Normal and LGD biopsies to characterize the architectural and cytological properties of specimens based on the computational analysis. We produced two sets of enhanced images. Then, by considering the fractal concept, we defined a new scale-dependent function in the generalized fractal dimension formulation to include the special information of both preprocessed images together. Then, for each image, a pattern was computed from variations of tissue fractal geometry across the pathway of dysplasia development. We proposed features extracted from these patterns to classify deviations of tissue characteristics from the normal stage. This method successfully differentiated the two diagnosis classes with statistical significance and high performance (accuracy = 97.78% ± 0.05, p

Mo1748 - Validation of Novel Methylated DNA Markers for the Detection of Esophageal Squamous Cell Carcinoma and Dysplasia: Multi-National Tissue Study

Mo1748 - Validation of Novel Methylated DNA Markers for the Detection of Esophageal Squamous Cell Carcinoma and Dysplasia: Multi-National Tissue Study

Diagnosis and therapy of esophageal squamous cell dysplasia and early esophageal squamous cell cancer

Diagnosis and therapy of esophageal squamous cell dysplasia and early esophageal squamous cell cancer

540 Biomarkers for Early Detection of Esophageal Squamous Cell Dysplasia and Carcinoma

540 Biomarkers for Early Detection of Esophageal Squamous Cell Dysplasia and Carcinoma

Multi-susceptibility genes associated with the risk of the development stages of esophageal squamous cell cancer in Feicheng County

BackgroundThe purpose of this study was to evaluate the association of multi-genotype polymorphisms with the stepwise progression of esophageal squamous cell cancer (ESCC) and the possibility of predicting those at higher risk.MethodsA total of 1,004 subjects were recruited from Feicheng County, China, between Jan. 2004 and Dec. 2007 and examined by endoscopy for esophageal lesions. These subjects included 270 patients with basal cell hyperplasia (BCH), 262 patients with esophageal squamous cell dysplasia (ESCD), 226 patients with ESCC, and 246 controls with Lugol-voiding area but diagnosed as having normal esophageal squamous epithelial cells by histopathology. The genotypes for CYP2E1 G1259C, hOGG1 C326G, MTHFR C677T, MPO G463A, and ALDH2 allele genes were identified in blood samples collected from all participants.ResultsThe alleles ALDH2 and MTHFR C677T were critical for determining individual susceptibility to esophageal cancer. Compared to the ALDH 1*1 genotype, the ALDH 2*2 genotype was significantly associated with increased risks of BCH, ESCD, and ESCC. However, the TT genotype of MTHFR C677T only increased the risk of ESCC. Further analysis revealed that the combination of the high-risk genotypes 2*2/1*2 of ALDH 2 and TT/TC of MTHFR C677T increased the risk of BCH by 4.0 fold, of ESCD by 3.7 fold, and ESSC by 8.72 fold. The generalized odds ratio (ORG) of the two combined genotypes was 1.83 (95%CI: 1.55-2.16), indicating a strong genetic association with the risk of carcinogenic progression in the esophagus.ConclusionsThe study demonstrated that the genotypes ALDH2*2 and MTHFR 677TT conferred elevated risk for developing esophageal carcinoma and that the two susceptibility genotypes combined to synergistically increase the risk.

Esophageal Squamous Cell Dysplasia and Delayed Differentiation With Deletion of Krüppel-Like Factor 4 in Murine Esophagus

Krüppel-like factor 4 (Klf; previously known a gut-enriched Krüppel-like factor) is a DNA-binding transcriptional regulator highly expressed in skin and gastrointestinal epithelia, specifically in regions of cellular differentiation. Homozygous null mice for Klf4 die shortly after birth from skin defects, precluding their analysis at later stages. The aim of this study was to analyze the function of Klf4 in keratinocyte biology and epithelial homeostasis in the adult by focusing on the squamous lined esophagus. By using the ED-L2 promoter of Epstein-Barr virus to drive Cre, we obtained tissue-specific ablation of Klf4 in the squamous epithelia of the tongue, esophagus, and forestomach. Mice with loss of Klf4 in esophageal epithelia survived to adulthood, bypassing the early lethality. Tissue-specific Klf4 knockout mice had increased basal cell proliferation and a delay in cellular maturation; these mice developed epithelial hypertrophy and subsequent dysplasia by 6 months of age. Moreover, loss of Klf4 in vivo was associated with increased expression of the pro-proliferative Klf5, and Klf4 down-regulated Klf5 both transcriptionally and posttranscriptionally. By using gene expression profiling, we also showed decreased expression of critical late-stage differentiation factors and identified alterations of several genes important in cellular differentiation. Klf4 is essential for squamous epithelial differentiation in vivo and interacts with Klf5 to maintain normal epithelial homeostasis.

Association of genetic polymorphisms of aldehyde dehydrogenase-2 with esophageal squamous cell dysplasia

To demonstrate the possible associations between genetic polymorphisms of aldehyde dehydrogenase-2 (ALDH2) and esophageal squamous cell dysplasia (ESCD). All participants came from an area of high incidence of esophageal cancer and underwent an endoscopic staining examination; biopsies were taken from a non-staining area of the mucosa and diagnosed by histopathology. Based on the examinations, the subjects were divided into the control group with normal esophageal squamous epithelial cells and the ESCD group. ALDH2 genotypes of 396 cases were determined including 184 ESCD cases and 212 controls. The odds ratio (OR) and 95% confidence intervals (95% CI) were calculated by binary logistic regression models. The distribution of ALDH2 genotypes showed significant differences between the two groups. The adjustment factors were gender and age in the logistic regression models. Compared with 2*2/2*2 genotype, 2*1/2*1 genotype was found to be a risk factor for ESCD, and the OR (95% CI) was 4.50 (2.21-9.19). There were significant correlations between ALDH2 genotypes and alcohol drinking/smoking/history of esophageal cancer. The ALDH2 polymorphism is significantly associated with ESCD.

Association of serum levels of lipid and its novel constituents with the different stages of esophageal carcinoma.

BackgroundThe aim of the study was to evaluate the association of immunoglobulin G type of autoantibodies to oxidized low-density lipoprotein (oxLDL-lgG) and oxLDL-lgM with the progression of esophageal squamous cell carcinoma (ESSC).MethodsResidents from Feicheng, China aged 40 to 69 years were screened for esophageal lesions in a screening program conducted during the period of January 2008 to December 2006. There were 33 controls with normal esophageal squamous epithelium cells, 37 patients with basal cell hyperplasia, 47 with esophageal squamous cell dysplasia, and 43 with ESCC. All the participants were diagnosed by biopsy and histopathological examination. Adiponectin, oxidized low-density lipoprotein (oxLDL), autoantibodies against oxLDL (oxLDL-ab), OxLDL-lgG, and OxLDL-lgM were determined by enzyme linked immunosorbent assay (ELISA). Total cholesterol, High-density lipoprotein (HDL), triglyceride, serum albumin, and blood pressure were co-estimated. Analysis of covariance for lipid levels was used to control the influence of covariates.ResultsThe level of oxLDL-lgM increased gradually along with esophageal carcinoma progression. The oxLDL-lgM levels in the ESCC group were the highest after possible covariates were controlled. Binary logistic regression showed that oxLDL-lgM had a positive correlation with the development of esophageal carcinoma, while oxLDL and oxLDL-ab had a negative correlation with ESSC. No significant association between the levels of oxLDL-lgG and adiponectin and the different stages of ESSC was observed.ConclusionThe present study shows that the decreased oxLDL and oxLDL-ab and the elevated oxLDL-lgM serum levels may relate to the development and progression of ESSC.

Toll‐like receptor 9 (TLR9) expression is correlated to progression of esophageal squamous cell carcinoma

Toll‐like receptors (TLR) identify a wide variety of microbial ligands and endogenous DNA fragments from apoptotic cells. TLR9 is located in endosomes and it recognises CpG‐DNA. TLR9 activation induces cell invasion in a model of breast cancer, suggesting a role for infections or tissue lysis in the pathophysiology behind cancer invasion. In this study we describe an expression pattern of TLR9 in esophagus samples varying from esophageal squamous cell dysplasia to squamous cell carcinoma. We have found strong positive staining in dysplastic samples which is differently organised to that of the healthy controls. Our data clearly indicates that TLR9‐expression, in addition of its role in immunity, is a marker of differentiation, proliferation or invasiveness in esophagus carcinoma sequence. This interpretion is supported by strong basal expression of TLR9 in the esophageal epithelium, where the stem cells are thought to reside, and weakening of the expression towards the lumen, where peeling off of the epithelia takes place. Also in the neoplastic changes, the epithelial expression of TLR9 becomes very prominent. This new staining might enable specific early recognition of malignant transformation and be a basis for new kind of understanding in the mucosal immunology and cancer. This study was supported by Finnish Medical Foundation and Oulu University Hospital grants.

Relationship Between Oxidized LDL Antibodies and Different Stages of Esophageal Carcinoma

The aim of the study was to evaluate the association of antibodies against oxidized low-density lipoproteins (oxLDL-Ab) with esophageal carcinogenic progression. All 40- to 69-year-old residents from Feicheng were screened for esophageal lesions by endoscopic staining with 1.2% iodine solution combined with pathological evaluations. In this study there were 33 controls with normal esophageal squamous epithelium cells, 37 patients with basal cell hyperplasia, 47 with esophageal squamous cell dysplasia, and 43 with esophageal squamous cell carcinoma (ESCC). OxLDL-Ab was determined by ELISA. Total cholesterol (TC), high-density lipoproteins (HDL), triglycerides, serum albumin and blood pressure were co-estimated. Analysis of covariance (ANCOVA) was used when comparing oxLDL-Ab among the four groups to control the influence of covariates. Cumulative logistic model was applied to study the influencing factors for the multistage development of esophageal carcinoma. The level of oxLDL-Ab decreased gradually along with the different stages of esophageal carcinogenic progression, with the ESCC group being the lowest after controlling for possible covariates. Cumulative logistic model showed that oxLDL-Ab had a negative correlation with the development of esophageal carcinoma. LDL, HDL, and TC were also decreased in patients with ESCC. Antibodies against oxLDL were decreased in patients with esophageal carcinoma. Although the unambiguous role of oxLDL-Ab needs further studies to elucidate, the results may give us some insight in the research of etiological factors for esophagael cancer in the future.

Association of the Polymorphisms of MTHFR C677T, VDR C352T, and MPO G463A with Risk for Esophageal Squamous Cell Dysplasia and Carcinoma

From January 2004 to December 2006 a program of endoscopic screening for esophageal lesions was carried out in the high incidence area of esophageal cancer in Feicheng County, China. It provided the samples to evaluate the association of polymorphisms of methylenetetrahydrofolate reductase (MTHFR) C677T, vitamin D receptor (VDR) C352T, and myeloperoxidase (MPO) G463A genotypes with esophageal squamous cell dysplasia and carcinoma. The subjects in the study were divided into 127 dysplasia cases, 126 squamous cell carcinoma cases, and 169 normal controls. Analyses of the MTHFR C677T, VDR C352T, and MPO G463A genotypes were performed using the PCR-restriction fragment length polymorphism (RFLP) method, whereas the multinomial logistic regression model was used in the data analysis to assess the odds ratios (ORs) related to dysplasia and carcinoma. Compared with the CC genotype of MTHFR C677T, the TT/TC of the genotype significantly increased the risk of the esophageal squamous cells dysplasia [OR, 2.25; 95% confidence interval (CI), 1.18-4.31]; the OR of esophageal squamous cancer was 1.58 (95% CI, 0.85-2.97) after adjustments for age, sex, and years of education. There was an interaction between the TT/TC genotype and alcohol drinking, smoking, and family history of esophageal cancer in the risk of esophageal dysplasia and carcinoma. Neither the VDR C352T nor the MPO G463A genotype had manifested association with the dysplasia and carcinoma of the disease, whereas the MTHFR 677TT genotype may be a genetic risk factor for esophageal dysplasia and carcinoma.

Differential protein expression between esophageal squamous cell carcinoma and dysplasia, and prognostic significance of protein markers

The aim of the current study was to evaluate the protein expression involved in the progression from dysplasia to invasive esophageal squamous cell carcinomas and to analyze the prognostic value of markers. Immunohistochemistry was performed for cell cycle regulators [p53, p21, p27, p16, cyclin D1, Rb], apoptosis-related proteins [Fas, Fas-L, FADD, TRAIL, DR4, DR5, caspase-8, caspase-3, bcl-2, Bax], tumor suppressor proteins [ β-catenin, E-cadherin, FHIT, Smad 4, VHL, PTEN, KAI-1], and oncoproteins [c-myc, COX-2, EGFR]. Caspase-3, TRAIL, Fas-L, Fas, Smad 4, VHL, E-cadherin, and EGFR revealed significant differences between dysplasia and their corresponding invasive cancer portion in 25 cases. In a total of 118 cases of invasive cancer, proteins with frequent (⩾60% of the cases) alterations were p53 (overexpression in 64% of SCCs), p27 (loss in 91%), p16 (loss in 81%), and FHIT (loss in 75%). Early clinical stage and bcl-2 immunopositivity were related to the survival rate of patients. In conclusion, caspase-3, TRAIL, Fas-L, Fas, Smad 4, VHL, E-cadherin, and EGFR may be involved in the progression from dysplasia to invasive esophageal SCCs. Clinical stage and bcl-2 are independent prognostic factors throughout the multivariate analysis.

Utility of Chromoendoscopy with Lugol Solution Spraying in Early Detection of Esophageal Squamous Cell Carcinoma and Dysplasia

Utility of Chromoendoscopy with Lugol Solution Spraying in Early Detection of Esophageal Squamous Cell Carcinoma and Dysplasia

Expression of p53-related protein p63 in the gastrointestinal tract and in esophageal metaplastic and neoplastic disorders

p63 is a p53-related DNA-binding protein that helps regulate differentiation and proliferation in epithelial progenitor cells. Its expression has never been evaluated in the human gastrointestinal tract. The aim of this study was to evaluate the expression of p63 in the esophagus and related metaplastic and neoplastic disorders to gain insight into the pathogenesis of these processes. Of particular interest was the expression of p63 in Barrett esophagus (BE) and in BE-associated multilayered epithelium. Multilayered epithelium has been postulated to represent an early precursor to the development of BE primarily because it shares morphologic and immunophenotypic features of both squamous and columnar epithelium, and has been shown prospectively to be highly associated with BE. Routinely processed mucosal biopsy or resection specimens that contained normal esophageal squamous epithelium (n = 20), squamous dysplasia (n = 4), squamous cell carcinoma (n = 7), BE (n = 10), BE-associated multilayered epithelium (n = 13), esophageal mucosal gland ducts (n = 10), BE-associated dysplasia (n = 12), and BE-associated adenocarcinoma (n = 7) were immunostained for p63 to determine the extent and location of staining. p63 staining was compared with the staining patterns observed for p53, Ki 67 (proliferation marker), and cytokeratins (CKs) 13 (squamous marker), 14 (basal squamous marker), 8/18 (columnar marker), and 19 (basal/columnar marker). Expression of p63 messenger RNA (mRNA) isoforms was also analyzed by reverse-transcription polymerase chain reaction of freshly isolated tissues. In the normal esophagus, p63 was expressed in the basal and suprabasal layers of the squamous epithelium and in basal cells that line the mucosal gland ducts but was negative in all other epithelia of the gastrointestinal tract, including the stomach, small intestine, and colon. Similarly, p63 was not expressed in BE, but it, was present in the basal layer of multilayered epithelium in 9 of 13 cases (69%). p63-positive cells in multilayered epithelium and in the mucosal gland duct epithelium were positive for CK8/18 (100%) and CK13 (67% and 30%, respectively) and negative for CK14 (0%), in contrast to p63-positive cells in squamous epithelium, which were positive for CK14 and CK13 (100%) but negative for CK8/18. In neoplastic tissues, p63 was diffusely expressed in all cases of esophageal squamous cell dysplasia and carcinoma but was negative in all cases of esophageal and colorectal adenocarcinoma. The ΔN isoform of p63 mRNA predominated in all benign and neoplastic squamous tissues examined. p63 may represent a marker of 2 distinct epithelial progenitor cells (basal squamous epithelium and gland duct epithelium) in the esophagus. P63 is upregulated in squamous neoplastic conditions and in this manner may play a role in squamous carcinogenesis. These data also indicate that multilayered epithelium is phenotypically similar to, and may share a lineage relationship with, mucosal gland duct epithelium. HUM PATHOL 32:1157-1165. Copyright © 2001 by W.B. Saunders Company

Natural history and significance of esophageal squamous cell dysplasia

Balloon-mesh cytologic screening for esophageal cancer done in 255 asymptomatic high-risk United States veterans (age greater than 40 years, ethanol abuse for greater than 20 years, and cigarette smoking greater than 20 pack years) identified 37 patients with squamous cell dysplasia. Of the 37 patients with dysplasia, 28 were re-evaluated prospectively at 6-month intervals for up to 36 months by balloon-mesh cytology, esophagoscopy with vital staining and biopsies, chest radiographs, oropharyngeal examination, and indirect laryngoscopy. During prospective follow-up evaluation, cytology specimens were repetitively normal in 16 patients (57%), showed inflammatory changes in eight patients (29%), persisted as dysplasia in two patients (7%) (both had endoscopic and histologic evidence of esophagitis), and progressed to carcinoma in two patients (7%) (one esophageal, one laryngeal). Although histologic findings concurred with the resolution of dysplasia, biopsy specimens were characterized by a similar difficulty in distinguishing dysplasia from inflammation. Erroneous histologic diagnoses of carcinoma in situ were made in two patients with reflux esophagitis evident endoscopically and confirmed during the course of a 24-36 month follow-up period. The authors conclude that squamous cell dysplasia detected by balloon-mesh cytology is seldom a precursor of esophageal cancer in the high-risk U.S. population but, rather, is often related to esophagitis. Thus, balloon-mesh cytology has limited use as a screening method for the early detection of esophageal cancer in the United States.