Meta-analysis of ADH1B and ALDH2 polymorphisms and esophageal cancer risk in China

AIM: To evaluate whether alcohol dehydrogenase-1B (ADH1B) His47Arg and aldehyde dehydrogenase-2 (ALDH2) Glu487Lys polymorphism is involved in the esophageal squamous cell carcinoma (ESCC) risk in Chinese Han population. METHODS: Seven studies of ADH1B and ALDH2 genotypes in Chinese Han population in 1450 cases and 2459 controls were included for meta-analysis. Stratified analyses were carried out to determine the genealcohol and gene-gene interaction with ESCC risk. Potential sources of heterogeneity betwee...     

Identification of potential core genes in esophageal carcinoma using bioinformatics analysis

Background:Esophageal squamous cell carcinoma (ESCC) is a common human malignancy worldwide. The tumorigenesis mechanism in ESCC is unclear.Materials and methods:To explore potential therapeutic targets for ESCC, we analyzed 3 microarray datasets ({"type":"entrez-geo","attrs":{"text":"GSE20347","term_id":"20347"}}GSE20347, {"type":"entrez-geo","attrs":{"text":"GSE38129","term_id":"38129"}}GSE38129, and {"type":"entrez-geo","attrs":{"text":"GSE67269","term_id":"67269"}}GSE67269) derived from the gene expression omnibus (GEO) database. Then, the GEO2R tool was used to screen out differently expressed genes (DEGs) between ESCC and normal tissue. Gene ontology function and kyoto encyclopedia of genes and genomes pathway enrichment analysis were performed using the database for annotation, visualization and integrated discovery to identify the pathways and functional annotation of DEGs. Protein–protein interaction of these DEGs was analyzed based on the search tool for the retrieval of interacting genes database and visualized by Cytoscape software. In addition, we used encyclopedia of RNA interactomes (ENCORI), gene expression profiling interactive analysis (GEPIA), and the human protein atlas to confirm the expression of hub genes in ESCC. Finally, GEPIA was used to evaluate the prognostic value of hub genes expression in ESCC patients and we estimated the associations between hub genes expression and immune cell populations (B Cell, CD8⁺ T Cell, CD4⁺ T Cell, Macrophage, Neutrophil, and Dendritic Cell) in esophageal carcinoma (ESCA) using tumor immune estimation resource (TIMER).Results:In this study, 707 DEGs (including 385 upregulated genes and 322 downregulated genes) and 6 hub genes (cyclin B1 [CCNB1], cyclin dependent kinase 1 [CDK1], aurora kinase A [AURKA], ubiquitin conjugating enzyme E2C [UBE2C], cyclin A2 [CCNA2], and cell division cycle 20 [CDC20]) were identified. All of the 6 hub genes were highly expressed in ESCC tissues. Among of them, only CCNB1 and CDC20 were associated with stage of ESCC and all of them were not associated with survival time of patients.Conclusion:DEGs and hub genes were confirmed in our study, providing a thorough, scientific and comprehensive research goals for the pathogenesis of ESCC.     

Polymorphisms of the DNA repair gene xeroderma pigmentosum groups A and C and risk of esophageal squamous cell carcinoma in a population of high incidence region of North China

Aim Inherited polymorphisms of DNA repair genes may contribute to variations in DNA repair capacity (DRC) and genetic susceptibility to different cancers. The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of xeroderma pigmentosum group A (XPA) and XPC can influence the risk of esophageal squamous cell carcinoma (ESCC). Methods In this report, one SNP of XPA and three SNPs of XPC were genotyped by polymerase-chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) assay in 327 ESCC patients and 612 healthy controls in a high incidence region of North China. Results Family history of upper gastrointestinal cancers (UGIC) may increase the risk of developing ESCC. The overall genotype and allelotype distributions of XPA A23G in ESCC patients were significantly different from that in healthy controls (P < 0.05). The A/G + G/G genotype significantly decreased the risk of developing ESCC compared with A/A genotype. When stratified for family history of UGIC, compared with A/A genotype, A/G + G/G genotype significantly decreased the risk of ESCC in groups with negative history of UGIC. The overall genotype and allelotype distributions of XPC intron 9 PAT^+/- and exon 15 Lys939Gln and exon 8 Val499Ala in ESCC patients were not significantly different from that in healthy controls (P > 0.05). When stratified for smoking status and UGIC family history, compared with A/A genotype, C/C genotype of exon 15 Lys939Gln significantly increased the risk of developing ESCC in non-smoker group. Conclusions We concluded that XPA23 polymorphism may be useful markers for identifying individuals at risk of developing ESCC. C/C genotype of XPC exon 15 may be one of the factors that affect the risk of developing ESCC in nonsmoking population in the high incidence region of China.     

Risk appraisal and endoscopic screening for esophageal squamous cell carcinoma in Japanese populations

Epidemiological studies have shown several strong predictors for selecting Japanese persons at high risk for esophageal squamous cell carcinoma (ESCC). (1) Alcohol consumption and tobacco smoking synergistically increase the risk, and a low intake of green and yellow vegetables or fruit and a low body mass index also increase the risk of ESCC. (2) The presence of esophageal distinct iodine-unstained lesions and melanosis are associated with an increased risk of ESCC. (3) The combination of alcohol consumption and inactive heterozygous aldehyde dehydrogenase-2 (ALDH2) and less-active homozygous alcohol dehydrogenase-1B (ADH1B) increases the risk of ESCC in a multiplicative fashion. (4) The results of a simple flushing questionnaire predict the ALDH2 phenotype with a high accuracy. (5) High mean corpuscular volume (MCV), which is induced by heavy drinking, high acetaldehyde exposure, heavy smoking, and poor nutrition, may be useful in identifying high-risk persons. Endoscopic screening with esophageal iodine staining in Japanese high-risk populations yields very high rates of early ESCC. Treatment of early ESCC by endoscopic mucosectomy has become a widespread practice in Japan and has succeeded in improving the outcome of this high-mortality cancer. New evidence concerning ALDH2/ADH1B/alcohol flushing/MCV-related cancer susceptibility has renewed interest in alcohol and acetaldehyde as important subjects for cancer research and has served as a powerful tool for cancer prevention and cancer screening of Japanese subjects. Review articles on this topic also appeared in the previous issue (Volume 4 Number 3). An editorial related to this article is available at .     

Relationships of early esophageal cancer with human papillomavirus and alcohol metabolism

BACKGROUND It is well known that an alcohol consumption habit together with inactive heterozygous aldehyde dehydrogenase-2(ALDH2) is an important risk factor for the development of esophageal squamous cell carcinoma(ESCC). It remains controversial whether human papillomavirus(HPV) infection contributes to the occurrence/development of ESCC. There has been no study in which the relationship between ESCC and HPV in addition to alcohol dehydrogenase-1 B(ADH1 B) and ALDH2 genotypes was evaluated.AIM To evaluate relationships between HPV infection and development of esophageal cancer, particularly early esophageal cancer, based on ADH1 B/ALDH2 polymorphisms.METHODS We conducted an exploratory retrospective study using new specimens, and weenrolled 145 patients who underwent endoscopic resection for superficial ESCC and had been observed for more than two years by both physical examination and endoscopic examination in Hokkaido University Hospital. Saliva was collected to analyze genetic polymorphisms of ADH1 B/ALDH2. We performed in situ hybridization for resected specimens to detect HPV by using an HPV type 16/18 probe.RESULTS HPV was detected in 15(10.3%) of the 145 patients with ESCC. HPV-positive rates in inactive ALDH2*1/*2 and ALDH2*1/*1 + *2/*2 were 10.8% and 9.8%, respectively(P = 1.00). HPV-positive rates in slow-metabolizing ADH1 B*1/*1 and ADH1 B*1/*2 + *2/*2 were 12.0% and 10.0%, respectively(P = 0.72). HPV-positive rates in the heavy or moderate alcohol consumption group and the light or rare consumption group were 11.1% and 8.7%, respectively(P = 0.68). HPV-positive rates in the heavy smoking group and the light or no smoking group were 11.8% and 8.3%, respectively(P = 0.59). The 3-year incidence rates of secondary ESCC or head and neck cancer after initial treatment in the HPV-positive and HPVnegative groups were 14.4% and 21.4%(P = 0.22), respectively.CONCLUSION In the present situation, HPV status is considered to be less important than other risk factors, such as alcohol consumption, smoking habit, ADH1 B/ALDH2 polymorphisms, and HPV status would therefore have no effect on ESCC risk management.     

Genotypes of CYP1A1, SULT1A1 and SULT1A2 and risk of squamous cell carcinoma of esophagus: outcome of a case–control study from Kashmir,India

Studies on associations of various polymorphism in xenobiotic metabolizing genes with different cancers including esophageal squamous cell carcinoma (ESCC) are mixed and inconclusive. To evaluate the association of CYP1A1*4, SULT1A1*2 and SULT1A2*2 genotypes with ESCC risk and their modifying effects on different risk factors of ESCC, we conducted a case–control study in Kashmir, India, an area with relative high incidence of ESCC. We recruited 404 histopathologically confirmed ESCC cases, and equal number of controls, individually matched for sex, age and district of residence to respective case. Information was obtained on various dietary, lifestyle and environmental factors in face‐to‐face interviews, using a structured questionnaire, from each subject. Genotypes were analyzed by polymerase chain reaction, restriction fragment length polymorphism and direct sequencing. Conditional logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (95% CIs). A higher risk was observed in the subjects who harbored variant genotype of CYP1A1*4 (OR = 2.06; 95% CI: 1.28–3.32); and the risk was further enhanced in ever smokers (OR = 3.47; 95% CI: 1.62–7.42), adobe dwellers (OR = 6.71; 95% CI: 3.02–14.89), and biomass fuel users (OR = 5.11; 95% CI: 1.34–19.50). We did not find any significant differences in the polymorphic variants of SULT1A1*2 and SULT1A2*2 between cases and controls. The study indicates that, unlike SULT1A1*2 and SULT1A2*2, the polymorphism of CYP1A1*4 is associated with ESCC risk. However, replicative studies with larger sample size are needed to substantiate our findings.     

Comparative genomic hybridization analysis of genetic aberrations associated with development of esophageal squamous cell carcinoma in Henan, China

AIM： To characterize cytogenetic alterations in esophageal squamous cell carcinoma （ESCC） and its metastasis. METHODS： A total of 37 cases of primary ESCC and 15 pairs of primary ESCC tumors and their matched metastatic lymph nodes cases were enrolled from Linzhou, the high incidence area for ESCC in Henan, northern China. The comparative genomic hybridization （CGH） was applied to determine the chromosomal aberrations on the DNA extracted from the frozen ESCC and metastatic lymph node samples from these patients. RESULTS： CGH showed chromosomal aberrations in all the cases. In 37 cases of primary ESCC, chromosomal profile of DNA copy number was characterized by frequently detected gains at 8q （29/37, 78%）, 3q （24/37, 65%）, 5p （19/37, 51%）; and frequently detected losses at 3p （21/37, 57%）, 8p and 9q （14/37, 38%）. In 15 pairs of primary ESCC tumors and their matched metastatic lymph node cases, the majority of the chromosomal aberrations in both primary tumor and metastatic lymph node lesions were consistent with the primary ESCC cases, but new candidate regions of interest were also detected. The most significant finding is the gains of chromosome 6p with a minimum high-level amplification region at 6p12-6q12 in 7 metastatic lymph nodes but only in 2 corresponding primary tumors （P = 0.05） and 20p with a minimum high-level amplification region at 20p12 in 11 metastatic lymph nodes but only in 5 corresponding primary tumors （P 〈 0.05）. Another interesting finding is the loss of chromosome 10p and 10q in 8 and 7 metastatic lymph nodes but only in 2 corresponding primary tumors （P 〈 0.05）. CONCLUSION： Using the CGH technique to detect chromosomal aberrations in both the primary tumor and its metastatic lymph nodes of ESCC, gains of 8q, 3q and 5p and loss of 3p, 8p, 9q and 13q were specifically implicated in ESCC in Linzhou population. Gains of 6p and 20p and loss of 10pq may contribute to the lymph node metastasis of ESCC. These findings suggest that the gains and     

Plasma DNA methylation of Wnt antagonists predicts recurrence of esophageal squamous cell carcinoma

AIM:To detect the effects of plasma DNA methylation of Wnt antagonists/inhibitors on recurrence of esophageal squamous cell carcinoma(ESCC).METHODS:We used methylation-specific polymerase chain reaction to detect hypermethylation of the promoter of four Wnt antagonists/inhibitors(SFRP-1,WIF-1,DKK-3 and RUNX3) using DNA from the plasma of ESCC patients(n = 81) and analyzed the association between promoter hypermethylation of Wnt pathway modulator genes and the two-year recurrence of ESCC.RESULTS:Hypermethyla...     

No evidence of HPV DNA in esophageal squamous cell carcinoma in a population of Southern Brazil

AIM:To investigate the association between human papillomavirus(HPV)and esophageal squamous cell carcinoma(ESCC)in southern Brazil.METHODS:We studied 189 esophageal samples from125 patients from three different groups:(1)102 biopsies from 51 patients with ESCC,with one sample from the tumor and another from normal esophageal mucosa distant from the tumor;(2)50 esophageal biopsies from 37 patients with a previous diagnosis of head and neck squamous cell carcinoma(HNSCC);and(3)37 biopsies from esophageal mucosa with normal appearance from 37 dyspeptic patients,not exposed to smoking or alcohol consumption.Nested-polymerase chain reaction(PCR)with the MY09/11 and GP5/6 L1primers was used to detect HPV L1 in samples fixed in formalin and stored in paraffin blocks.All PCR reactions were performed with a positive control(cervicovaginal samples),with a negative control(Human Genomic DNA)and with a blank reaction containing all reagents except DNA.We took extreme care to prevent DNA contamination in sample collection,processing,and testing.RESULTS:The histological biopsies confirmed the diagnosis of ESCC in 52 samples(51 from ESCC group and 1 from the HNSCC group)and classified as well differentiated(12/52,23.1%),moderately differentiated(27/52,51.9%)or poorly differentiated(7/52,13.5%).One hundred twenty-eight esophageal biopsies were considered normal(51 from the ESCC group,42 from the HNSCC group and 35 from dyspeptic patients).Nine had esophagitis(7 from the HNSCC and 2 from dyspeptic patients).Of a total of 189 samples,only 6 samples had insufficient material for PCR analysis:1 from mucosa distant from the tumor in a patient with ESCC,3from patients with HNSCC and 2 from patients without cancer.In 183 samples(96.8%)GAPDH,G3PDH and/orβ-globin were amplified,thus indicating the adequacy of the DNA in those samples.HPV DNA was negative in all the 183 samples tested:52 with ESCC,9 with esophagitis and 122 with normal esophageal mucosa.CONCLUSION:There was no evidence of HPV infection in different ESCC from southern Br     

RASSF1A、MINT31启动子区高甲基化与食管上皮内瘤变和食管鳞癌的关系

背景：食管上皮内瘤变（EIN）是食管正常或慢性炎症组织向鳞癌转变过程中必经的病理阶段,目前对EIN的发生、发展尚缺乏临床实用的早期评估指标。目的：探讨肿瘤相关基因RASSF1A、MINT31启动子区高甲基化和DNA甲基转移酶1（DNMT1）表达与EIN和食管鳞癌发生、发展的关系。方法：以甲基化特异性PCR（MSP）检测30例食管鳞癌、80例EIN和20例正常食管组织中的RASSF1A、MINT31甲基化状态,以亚硫酸氢盐修饰后测序（BSP）验证MSP结果。以免疫组化方法检测EIN和食管鳞癌组织中的DNMT1蛋白表达。结果：正常食管组织、EIN、食管鳞癌组织中的RASSF1A和MINT31甲基化率分别为5．0％和5．0％、32．5％和26．2％、60．0％和46．7％,组间两两比较差异均有统计学意义（P〈0．05）。BSP方法证实。MSP示RASSFlA甲基化阳性的EIN和食管鳞癌组织。RASSF1A启动子区存在甲基化位点。低级别、高级别EIN和食管鳞癌组织中的DNMT1蛋白表达阳性率分别为32．5％、55．0％和76．7％,组间两两比较差异均有统计学意义（P〈0．05）。DNMT1高表达与RASSF1A甲基化相关,与MINT31甲基化无关。结论：RASSF1A、MINT31启动子区高甲基化和DNMT1表达上调在EIN和食管鳞癌的发生、发展中起重要作用,其检测或许有助于EIN的评估。     

PTEN hypermethylation profiles of Chinese Kazakh patients with esophageal squamous cell carcinoma

Aberrant DNA methylation of promoter region CpG islands may serve as an alternative mechanism to genetic defects in the inactivation of tumor suppressor genes (TSGs) in human malignancies. The aim of this study was to examine the promoter methylation status of the PTEN TSG and its association with esophageal squamous cell carcinoma (ESCC) carcinogenesis in a Chinese Kazakh population, which is known to have a relatively high ESCC incidence and mortality. The methylation status of the PTEN promoter region was determined in patients with ESCC (n = 95) and healthy individuals (n = 65) using highly sensitive Sequenom Epityper assays. The methylation level of the PTEN gene was significantly higher in patients with ESCC than in healthy controls. The median methylation level was 10.0% (interquartile range [IQR]: 7.0–11.0%) in patients with ESCC and 6.0% in controls (IQR: 4.0–9.0%; P = 0.001). PTEN methylation levels were higher in male patients with ESCC than in male controls, whereas a trend toward significance was observed between female patients with ESCC and female controls (P = 0.005 and P = 0.086, respectively). The PTEN methylation level was associated with histopathological grade and lymph node metastasis in patients with ESCC (P = 0.002 and P = 0.009, respectively). To our knowledge, this is the first report to show the presence of PTEN promoter CpG hypermethylation in ESCC and its association with tumor metastasis.     

Role of the Na~+/K~+/2Cl~- cotransporter NKCC1 in cell cycle progression in human esophageal squamous cell carcinoma

AIM:To investigate the role of Na+/K+/2Cl-cotransporter 1(NKCC1)in the regulation of genes involved in cell cycle progression and the clinicopathological significance of its expression in esophageal squamous cell carcinoma(ESCC).METHODS:An immunohistochemical analysis was performed on 68 primary tumor samples obtained from ESCC patients that underwent esophagectomy.NKCC1expression in human ESCC cell lines was analyzed by Western blotting.Knockdown experiments were conducted using NKCC1 small interfering RNA,and the effects on cell cycle progression were analyzed.The gene expression profiles of cells were analyzed by microarray analysis.RESULTS:Immunohistochemical staining showed that NKCC1 was primarily found in the cytoplasm of carcinoma cells and that its expression was related to the histological degree of differentiation of SCC.NKCC1 was highly expressed in KYSE170 cells.Depletion of NKCC1in these cells inhibited cell proliferation via G2/M phase arrest.Microarray analysis identified 2527 genes with altered expression levels in NKCC1depleted KYSE170.Pathway analysis showed that the top-ranked canonical pathway was the G2/M DNA damage checkpoint regulation pathway,which involves MAD2L1,DTL,BLM,CDC20,BRCA1,and E2F5.CONCLUSION:These results suggest that the expression of NKCC1 in ESCC may affect the G2/M checkpoint and may be related to the degree of histological differentiation of SCCs.We have provided a deeper understanding of the role of NKCC1 as a mediator and/or a biomarker in ESCC.     

Genomic characterization of esophageal squamous cell carcinoma: Insights from next-generation sequencing

Two major types of cancer occur in the esophagus: squamous cell carcinoma, which is associated with chronic smoking and alcohol consumption, and adenocarcinoma, which typically arises in gastric reflux-associated Barrett’s esophagus. Although there is increasing incidence of esophageal adenocarcinoma in Western counties, esophageal squamous cell carcinoma(ESCC) accounts for most esophageal malignancies in East Asia, including China and Japan. Technological advances allowing for massively parallel, high-throughput next-generation sequencing(NGS) of DNA have enabled comprehensive characterization of somatic mutations in large numbers of tumor samples. Recently, several studies were published in which whole exome or whole genome sequencing was performed in ESCC tumors and compared with matched normal DNA. Mutations were validated in several genes, including in TP53, CDKN2 A, FAT1, NOTCH1, PIK3 CA, KMT2 D and NFE2L2, which had been previously implicated in ESCC. Several new recurrent alterations have also been identified in ESCC. Combining the clinicopathological characteristics of patients with information obtained from NGS studies may lead to the development of effective diagnostic and therapeutic approaches for ESCC. As this research becomes more prominent, it is important that gastroenterologist become familiar with the various NGS technologies and the results generated using these methods. In the present study, we describe recent research approaches using NGS in ESCC.     

多种抑癌基因在食管鳞状细胞癌中的甲基化状态及其临床意义

抑癌基因甲基化与食管癌相关,目前多种抑癌基因与肿瘤家族史相关性的报道少见。目的：研究多种抑癌基因在食管鳞状细胞癌（ESCC）中的甲基化状态及其临床意义。方法：选取2010年2～7月浙江省肿瘤医院76例ESCC患者。应用MSP技术检测肿瘤组织和相应癌旁正常组织中APC、RARl32、CDHl、p16…、RASSFlA等5个抑癌基因的甲基化状态,并分析抑癌基因甲基化状态与肿瘤家族史的关系及其对预后的影响。结果：ESCC组织APC、RARe2、CDHl、p16慨、RASSFlA的甲基化率均显著高于相应癌旁正常组织（P〈0．05）。ESCC组织中APC、RARl32、CDHl、RASSFlA甲基化与肿瘤家族史相关（P〈0．05）：CDHl、RASSFlA甲基化患者的生存期明显低于非甲基化患者（P-0．015、P=0．016）。结论：ESCC患者存在抑癌基因APC、RARe2、CDHl、p16№、RASSFlA高甲基化;且APC、RARe2、CDHl、RASSFlA甲基化与肿瘤家族史显著相关,CDHl、RASSFlA甲基化患者的预后可能较差。     

RASSF1A gene promoter methylation in esophageal cancer specimens

SUMMARY. RASSF1A is frequently inactivated by promoter methylation in human cancers. To understand the involvement of the RASSF1A gene in esophageal squamous cell cancer (ESCC), we investigated the methylation of the RASSF1A gene in primary ESCC to define the frequency of this epigenetic aberration and its clinicopathological significance. Methylation-specific polymerase chain reaction (MSP) was used to detect RASSF1A gene methylation in DNA from 55 cases of ESCC. Methylation of the RASSF1A gene was found in 13 of 55 (24%) cases of primary ESCC. No association was found between the promoter methylation of the RASSF1A gene in primary ESCC and age, gender, localization, invasion depth, or tumor stage. Association was found with tumor differentiation. There was no correlation with its prognosis. In conclusion, it was suggested that an inactivation of the RASSF1A gene due to promoter methylation was associated with de-differentiation of the tumor in ESCC.     

Promoter methylation status of hMLH1, MGMT, and CDKN2A/p16 in colorectal adenomas

AIM:To investigate aberrant DNA methylation of CpG islands and subsequent low-or high-level DNA microsatellite instability(MSI)which is assumed to drive colon carcinogenesis. METHODS:DNA of healthy individuals,adenoma(tu-bular or villous/tubulovillous)patients,and colorectal carcinoma patients who underwent colonoscopy was used for assessing the prevalence of aberrant DNA methylation of human DNA mismatch repair gene mutator L homologue 1(hMLH1),Cyclin-dependent kinase inhibitor 2A(CDKN2A/p16),and O-6-methy...