Socioeconomic status and esophageal squamous cell carcinoma risk in Kashmir,India

Studies have persistently associated esophageal squamous cell carcinoma (ESCC) risk with low socioeconomic status (SES), but this association is unexplored in Kashmir, an area with a high incidence of ESCC in the northernmost part of India. We carried out a case–control study to assess the association of multiple indicators of SES and ESCC risk in the Kashmir valley. A total number of 703 histologically confirmed ESCC cases and 1664 controls matched to the cases for age, sex, and district of residence were recruited from October 2008 to January 2012. Conditional logistic regression models were used to calculate unadjusted and adjusted odds ratios and 95% confidence intervals. Composite wealth scores were constructed based on the ownership of several appliances using multiple correspondence analyses. Higher education, living in a kiln brick or concrete house, use of liquefied petroleum gas and electricity for cooking, and higher wealth scores all showed an inverse association with ESCC risk. Compared to farmers, individuals who had government jobs or worked in the business sector were at lower risk of ESCC, but this association disappeared in fully adjusted models. Occupational strenuous physical activity was strongly associated with ESCC risk. In summary, we found a strong relationship of low SES and ESCC in Kashmir. The findings need to be studied further to understand the mechanisms through which such SES parameters increase ESCC risk.     

Poor oral hygiene and risk of esophageal squamous cell carcinoma in Kashmir

Background:

Several studies have suggested an association between poor oral health and esophageal squamous cell carcinoma (ESCC). We conducted a case-control study in Kashmir, a region with relatively high incidence of ESCC in north India, to investigate the association between oral hygiene and ESCC risk.

Methods:

We recruited 703 histologically confirmed ESCC cases, and 1664 controls individually matched to the cases for age, sex, and district of residence. Conditional logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (CIs).

Results:

We found an inverse association between teeth cleaning and ESCC risk. As compared with never cleaning teeth, the OR (95% CI) was 0.41 (0.28–0.62) for cleaning less than daily and 0.44 (0.25–0.77) for cleaning at least once a day (P for trend=0.026) in models adjusted for multiple potential confounders, including several indicators of socioeconomic status. This association persisted after we limited our analyses to never tobacco users. The inverse association between cleaning teeth and ESCC was stronger with using brushes than with using sticks/fingers. We also found an association between the number of decayed, filled, and missing teeth and ESCC risk, but the trend of the associations was not statistically significant. Avoiding solid food and cold beverages because of teeth and oral problems were also associated with ESCC risk.

Conclusion:

We found an association between poor oral hygiene indicators and ESCC risk, supporting the previous studies that showed the same associations.     

Association of cyclin D1 gene polymorphisms with risk of esophageal squamous cell carcinoma in Kashmir Valley: a high risk area

Investigation of potential association of SNPs (G870A, rs9344; G1722C, rs678653) of cyclin D1 gene (CCND1) with susceptibility to esophageal squamous cell carcinoma (ESCC) in Kashmir valley (India). The study included 302 subjects comprising 151 ESCC cases and 151 controls. PCR‐RFLP and direct sequencing were employed for genotyping. The G870A polymorphism, the individuals carrying GA + AA genotype was having 2.80‐fold increased risk for development of ESCC (OR 2.8, 95% CI = 1.77–4.4; P = 0.0001) compared to GG genotype. Further a significantly higher risk was observed in individuals who consume >3 cups per day of salted tea (OR = 5.1; 95% CI = 1.6–16.7; P = 0.0016) and had smoking habits (OR = 6.3; 95% CI = 2.9–13.9; P = 0.0005). We also demonstrate for the first time in CCND1 1722 locus, the CC genotype was strongly associated with increased risk of developing ESCC (OR = 2.58; 95% CI = 1.61–4.15; P = 0.0001). In addition, the frequency of polymorphic C allele was also found to be higher in cases (OR = 1.92; 95% CI = 1.37–2.69; P = 0.0002). There appears to be an influence of CCND1 G870A/G1772C genotypes on genetic susceptibility to ESCC. Mol. Carcinog. ©2011 Wiley‐Liss, Inc.     

A variant in CYP26B1 associated with esophageal squamous cell carcinoma risk by affecting retinoic acid metabolism

All-trans retinoic acid (ATRA) is the natural and synthetic analogue of vitamin A, playing an essential tumor suppressive role in multiple cancers including the esophageal squamous cell carcinoma (ESCC). Cytochrome P450 family 26 subfamily B member 1 (CYP26B1) exerts a critical regulator of ATRA levels through specific inactivation of ATRA to hydroxylated forms. Our previous exome-wide analyses revealed a rare missense variant in CYP26B1 significantly associated with ESCC risk in the Chinese population. However, it is still unclear whether there are common variants in CYP26B1 affect the susceptibility of ESCC and the tumor promotion role of CYP26B1 in vivo. In this research, we conducted a two-stage case-control study comprised of 5057 ESCC cases and 5397 controls, followed by a series of biochemical experiments to explore the function of CYP26B1 and its common variants in the tumorigenesis of ESCC. Intriguingly, we identified a missense variant rs2241057[A>G] in the fourth exon of CYP26B1 significantly associated with the ESCC risk (combined odds ratio = 1.28; 95% confidence interval = 1.15–1.42; p = 2.96 × 10⁻⁶). Through further functional analysis, we demonstrated that ESCC cells with the overexpression of rs2241057[G] had a significant lower level of retinoic acid, compared with the overexpression of rs2241057[A] or the control vector. In addition, the CYP26B1 overexpression and knock-out ESCC cells affected cell proliferation rate both in vitro and in vivo. These results highlighted the carcinogenicity of CYP26B1 related to the ATRA metabolism in ESCC risk.     

Germline BRCA2 mutations and the risk of esophageal squamous cell carcinoma

The incidence of esophageal squamous cell carcinoma (ESCC) is very high among the Turkmen population of Iran. Family studies suggest a genetic component to the disease. Turkmen are ethnically homogenous and are well suited for genetic studies. A previous study from China suggested that BRCA2 might play a role in the etiology of ESCC. We screened for mutations in the coding region of the BRCA2 gene in the germline DNA of 197 Turkmen patients with ESCC. A nonsense variant, K3326X, was identified in 9 of 197 cases (4.6%) vs 2 of 254 controls (0.8%) (OR=6.0, 95% CI=1.3-28; P=0.01). This mutation leads to the loss of the C-terminal domain of the BRCA2 protein, a part of the region of interaction with the FANCD2 protein. We observed nine other BRCA2 variants in single cases only, including two deletions, and seven missense mutations. Six of these were judged to be pathogenic. In total, a suspicious deleterious BRCA2 variant was identified in 15 of 197 ESCC cases (7.6%).     

Identification of a novel SEREX antigen, SLC2A1/GLUT1, in esophageal squamous cell carcinoma

We have carried out SEREX (serological identification of antigens by recombinant cDNA expression cloning), and identified SLC2A1 (solute carrier family 2/facilitated glucose transporter, member 1) as an antigen recognized by serum IgG antibodies in patients with esophageal squamous cell carcinoma (SCC). The levels of serum anti-SLC2A1 antibodies (s-SLC2A1-Abs), examined by enzyme-linked immunosorbent assay using bacterially expressed glutathione-S-transferase-SLC2A1 fusion protein, were significantly higher in patients with esophageal SCC than in healthy donors. When using a cut-off level as the mean + 2x standard deviations of healthy donors, a total of 12 (21%) out of 57 SCC patients were revealed as positive for s-SLC2A1-Abs. The presence of s-SLC2A1-Abs was not associated with either clinicopathological factors or survival. Because s-SLC2A1-Abs were not associated with the positivity of other conventional serum markers, a combination assay of s-SLC2A1-Abs with these conventional serum markers may be useful for the diagnosis and monitoring of esophageal SCC.     

Sex and smoking differences in the association between gastroesophageal reflux and risk of esophageal squamous cell carcinoma in a high-incidence area: Golestan Cohort Study

Prior studies have conflicting findings regarding the association between gastroesophageal reflux disease (GERD) and esophageal squamous cell carcinoma (ESCC). We examined this relationship in a prospective cohort in a region of high ESCC incidence. Baseline exposure data were collected from 50 045 individuals using in-person interviews at the time of cohort entry. Participants were followed until they developed cancer, died, or were lost to follow up. Participants with GERD symptoms were categorized into any GERD (heartburn or regurgitation), mixed symptoms, or heartburn alone. Multivariable Cox regression was used to assess the relationship between GERD symptom group and histologically confirmed ESCC. The model was adjusted for known risk factors for GERD and ESCC. 49 559 individuals were included in this study, of which 9005 had GERD symptoms. Over 13.0 years of median follow up, 290 individuals were diagnosed with ESCC. We found no association between any GERD and risk of ESCC (aHR 0.90, 95% CI: 0.66-1.24, P = .54). Similar findings were observed for the GERD symptom subtypes. Significant interactions between any GERD and sex (P = .013) as well as tobacco smoking (P = .028) were observed. In post-hoc analyses, GERD was associated with a decreased risk of ESCC in men (aHR 0.51, 95% CI: 0.27-0.98 P = .04) and in smokers (aHR 0.26, 95% CI: 0.08-0.83 P = .02). While there was little evidence for an overall association between GERD symptoms and ESCC risk, significant interactions with sex and smoking were observed. Men and smokers with GERD symptoms had a lower risk of ESCC development.     

CIP2A is overexpressed in esophageal squamous cell carcinoma

A human oncoprotein-designed cancerous inhibitor of PP2A (CIP2A) has been recently identified, which can stabilize c-Myc protein by inhibiting its degradation mediated by protein phosphatase 2A (PP2A) in tumor cells and promote the proliferation of various cancer cells. Esophageal squamous cell carcinoma (ESCC) is a highly aggressive cancer with poor prognosis worldwide. However, the underlying molecular mechanism of the development of ESCC is still poorly understood. In the present study, the CIP2A expression between normal and malignant esophageal tissues was compared by immunohistochemical analysis; moreover, the mechanisms of CIP2A-mediated tumorigenesis were investigated by evaluating its role in cell proliferation, cell cycle, apoptosis and senescence. We found that the positive staining of CIP2A was found in 36 of 40 (90%) of cancer tissues, whereas only 8 of 40 (20%) normal esophageal mucosa exhibited positive CIP2A staining. The CIP2A is significantly overexpressed in human esophageal tumors when compared with normal tissues (χ² = 39.6, P < 0.01). On the other hand, the CIP2A expression was not associated with age, gender, tumor burden, or differentiation status. Depletion of CIP2A expression led to impaired clonogenicity and senescence, which is the primary mechanism of CIP2A in oncogenesis. Therefore, CIP2A may be a candidate in diagnosis and therapy of esophageal cancer.     

Notch1基因与食管鳞状细胞癌

研究证实 Notch1在食管鳞状细胞癌中呈低表达,起抑癌基因作用。研究 Notch1基因与淋巴结转移、生存率之间的关系可以为食管鳞状细胞癌的治疗及预后评估提供更多有价值的帮助。     

Polymorphisms of GSTP1 and GSTT1, but not of CYP2A6, CYP2E1 or GSTM1, modify the risk for esophageal cancer in a western population

Esophageal cancer is among the most common and fatal tumors in the world. Eighty percent of esophageal tumors are esophageal squamous cell carcinoma (ESCC). Brazil is one of the high incidence areas in the West, where tobacco and alcohol consumption have been associated with ESCC. However, polymorphisms in xenobiotic metabolizing genes may also contribute to the risk. Therefore, in this study, we analyzed the risk of ESCC associated with tobacco and alcohol consumption and with polymorphisms of CYP2A6 (CYP2A6*2), CYP2E1 (CYP2E1*5B, CYP2E1*6), GSTP1 (Ile105Val), GSTM1 and GSTT1 null genotypes in 126 cases and 252 age- and gender-matched controls. Data on the amount, length and type of tobacco and alcohol consumed were collected, and DNA was extracted from blood lymphocytes from all individuals. Polymorphisms were analyzed by polymerase chain reaction (PCR)-multiplex (GSTM1 and T1), PCR-Restriction Fragment Length Polymorphism (CYP2E1*5B and *6 and GSTP1 Ile105Val) or allele-specific PCR amplification (CYP2A6*2). Risks were evaluated by multivariate conditional regression analysis. As expected, tobacco [odds ratio (OR) = 6.71, 95% confidence interval (95% CI) 3.08-14.63] and alcohol (OR = 16.98, CI 7.8-36.98) consumption, independently or together (OR = 26.91, CI 13.39-54.05) were risk factors. GSTP1 Ile105Val polymorphism was an independent risk factor (OR = 2.12, CI 1.37-3.29), whereas GSTT1 wild-type was an independent protective factor for ESCC (OR = 0.37, CI 0.16-0.79). There was approximately 80% statistical power to detect both results. There was no risk associated with CYP2A6, CYP2E1 and GSTM1 polymorphisms. In conclusion, this study suggests an opposite role of GSTP1 and GSTT1 polymorphisms for the risk for ESCC.     

Relevance of serum estradiol and estrogen receptor beta expression from a high-incidence area for esophageal squamous cell carcinoma in China

The striking 3-4:1 male predominance of esophageal squamous cell carcinoma (ESCC) has not yet been well explained. Our hypothesis is that the changes in level of estrogen and/or subtype of estrogen receptor (ER) may exert a protective factor in esophageal carcinogenesis and prognosis of ESCC. Radioimmunoassay (RIA) was used to determine the serum level of estradiol in healthy cohort from high-incidence area (HIA) and low-incidence area (LIA) for esophageal cancer as well as patients with ESCC from HIA in Henan, northern China. The ERβ expression profiling during the multi-stage progression of ESCC pathogenesis was evaluated by immunohistochemistry (IHC). Both males and females from HIA had significant decreases of serum estradiol in high-risk subjects predisposing for ESCC compared to healthy counterparts from LIA (P < 0.01). Furthermore, patients with ESCC from HIA developed the lowest level of estradiol (P < 0.01). ERβ expressed in precursor lesions of ESCC and changed quantitatively and qualitatively with disease progression during the multi-stages process of esophageal carcinogenesis. High frequency of ERβ expression was correlated with less aggressive potential of clinical behavior (P = 0.012, 0.015 for lymph node metastasis and tumor stage, respectively). This study indicates that lower serum level of estradiol may represent higher predisposition for development of ESCC, and ERβ expression and/or nuclear location may predict better outcome for patients with ESCC. The present results provide clues to explain the striking gender difference for ESCC, which warrants further investigations on potential applications of estrogen or analogs in prevention of ESCC.     

A novel gene,NMES1, downregulated in human esophageal squamous cell carcinoma

To isolate genes with different expression levels in human esophageal SCC, SSH and reverse Northern were performed between cancer tissue and its normal counterpart. Among the differentially expressed genes identified, we report here cDNA corresponding to a 0.88 kb mRNA (NMES1), whose expression was observed in all 36 adjacent normal esophageal mucosae, while 31 (86%) matched cancer tissues showed a marked reduction or complete lack of its expression. Sequence analysis of its full-length cDNA revealed a gene encoding a predicted polypeptide of 9 kDa. Northern blot showed that NMES1 was distributed mainly in the alimentary tract. The gene was mapped to 15q21.1 by screening the GenBridge 4 RH panel. Immunohistochemistry confirmed the downregulation of NMES1 in esophageal SCC at the protein level and showed that it is a nuclear protein. In situ hybridization revealed its different expression during mouse embryonic development, especially in bone, brain, stomach and intestine. The negative correlation of NMES1 expression with esophageal oncogenesis suggests its suppressive role in tumorigenesis of the esophagus, while the precise function of NMES1 still needs further investigation.     

食管鳞癌组织中COL1A2基因的表达与临床病理特征的相关性研究

目的探讨Ⅰ型胶原α2（COL1A2）和鳞状细胞癌相关抗原(SCCAg)与临床病理特征的关系。方法TIMER 20和GEPIA数据库分析COL1A2在肿瘤及食管癌中的表达,化学发光法检测血清中SCCAg的水平,免疫组化法检测食管鳞癌组织和癌旁正常组织中COL1A2蛋白的表达。生存分析采用GEPIA数据库,基因集富集分析采用GSEA数据库。结果GEPIA数据库分析显示,食管鳞癌中COL1A2的表达高于正常组织（P<005）,食管鳞癌TNM分期及与COL1A2的表达相关（P<005）。SCCAg在食管鳞癌患者中的阳性率为486%,其在高分化食管鳞癌患中表达增高（P<005）。COL1A2蛋白在食管鳞癌中强阳性率为735%,显著高于癌旁组织56%（P<005）。COL1A2过表达与食管鳞癌分化程度及其临床分期相关（P<005）。GEPIA数据库分析显示高表达COL1A2的患者无瘤生存率显著降低（P<005）。GSEA分析表明差异基因主要集中在粘着斑通路、细胞外基质结构组分、内质网内腔、细胞外结构组织。结论血清SCCAg的高表达对食管鳞癌诊断及其分化程度预测有一定的应用价值,而食管鳞癌组织COL1A2的高表达与食管鳞癌的进展及预后不良有关,对食管鳞癌的预后的判断有一定的临床参考价值,有望成为食管鳞癌新的治疗靶点。     

食管鳞癌组织HPV的检测及其意义的研究

目的:探讨人乳头状瘤病毒(HPV)与食管鳞癌的相关性,进一步明确人乳头瘤痛毒感染在食管癌病因学中的作用.方法:采用可检测23种HPV基因型的基因芯片和实时荧光定量PCR检测方法检测140例新鲜食管鳞癌组织的HPV型别,同时对照检测85例宫颈鳞癌组织中HPV的感染率.结果:85例宫颈鳞癌组织中HPV的阳性率为95.29%(81/85),共检测到9种HPV基因型,分别为HPV16、18、45、33、58、59、73、31和56,均为高危型感染.其中HPV16最常见,检出率达72.9%(62/85),其次是HPV18为16.5%(14/85),其他7型占28.2%(24/85),HPV双重感染检出率为11.8%(10/85).而在140例食管鳞癌组织中,未检测到任何基因型的HPV.结论:HPV感染似乎可能与高发区食管鳞癌的发生无关.     

血管内皮生长因子、血管生成素在食管鳞状细胞癌组织中的表达及其意义

目的 研究血管内皮生长因子(VEGF)、血管生成素(Ang)-1及Ang-2在食管鳞状细胞癌(ESCC)中的表达及其与预后的关系.方法 用免疫组织化学二步法检测60份ESCC、20份食管癌旁组织及10份正常食管黏膜上皮组织中VEGF、Ang-1及Ang-2的表达,计数微血管密度(MVD).结果 ESCC与食管癌旁组织及食管正常鳞状上皮组织比较,Ang-1表达率显著降低,而VEGF及Ang-2表达率则显著升高(P＜0.05).MVD在Ang-1阴性表达组显著高于阳性表达组(P=0.000);MVD在VEGF、Ang-2阳性表达组显著高于阴性表达组(P＜0.05).Ang-1表达与肿瘤分化程度显著相关(P=0.013).T3、T4期VEGF和Ang-2表达率显著高于T1、T2期(P＜0.05);VEGF及Ang-2在有淋巴结转移者显著高于无淋巴结转移者(P＜0.001).VEGF及Ang-2表达阳性组术后3年生存率低于阴性表达组(P＜0.05),Ang-1表达阳性和阴性组比较,差异无统计学意义(P=0.749).结论 Ang-1表达的降低及Ang-2、VEGF表达的增高可能与食管癌的发生、发展密切相关;VEGF及Ang-2在食管癌中表达提示预后不良.     

Expression of NDRG2 in esophageal squamous cell carcinoma

N‐Myc downstream‐regulated gene 2 (NDRG2), a new member of the N‐Myc downstream‐regulated gene family, has been found to be a differentially expressed gene involved in a variety of cancers. The present study aimed to investigate the expression of NDRG2 in esophageal squamous cell carcinoma (ESCC). Immunohistochemistry was performed in 154 samples from patients with ESCC to detect the expression level of NDRG2 and C‐MYC. Results indicated that the expression level of NDRG2 in the cancer samples was significantly lower than that in normal tissues; the trend of C‐MYC was the reverse. The Wilcoxon–Mann–Whitney test showed significant difference in the expression of NDRG2 in patients with different T stage, TNM stage, and differentiation degree of cancers (P = 0.036, 0.031, 0.001, respectively). Patients in stages I and II were followed up for 5 consecutive years and Kaplan–Meier survival analysis demonstrated that the survival time of ESCC patients with high expression of NDRG2 was longer than those with low expression during the 5‐year follow‐up period (P = 0.0018). Cox regression analysis indicated that low expression of NDRG2, cancer stage of pT1, and distant organ metastasis (pM1) were the independent poor prognostic factors of ESCC (P = 0.004, 0.019, 0.0013, respectively). Furthermore, up‐regulation of NDRG2 was introduced to ESCC cell lines (EC9706 and EC109) by plasmid transfection. In vivo and in vitro studies indicated that overexpression of NDRG2 markedly reduced proliferation and promoted the apoptosis of EC9706 and EC109 cells. In summary, our results demonstrated that NDRG2 played an important role in the proliferation of ESCC cells and the expression of NDRG2 in ESCC was closely related with the prognosis. (Cancer Sci 2010; 101: 1292–1299)     

Promoter hypermethylation of RASSF1A in esophageal squamous cell carcinoma.

PURPOSE: The RAS association domain family 1A (RASSF1A) gene, a candidate tumor suppressor gene, is frequently inactivated by hypermethylation of its promoter region in several human cancers. The aim of this study was to evaluate the promoter methylation status of the RASSF1A in esophageal squamous cell carcinoma. EXPERIMENTAL DESIGN: We analyzed the methylation status of RASSF1A promoter by methylation-specific PCR in 23 esophageal squamous cell carcinoma cell lines and 48 primary tumors. RESULTS: Hypermethylation of RASSF1A was found in 74% of cell lines and 52% of primary tumors. The presence of hypermethylation was statistically associated with loss of RASSF1A mRNA expression in both cell lines (P = 0.007) and primary tumors (P = 0.003). There was a statistically significant correlation between the presence of hypermethylation and tumor stage (P = 0.009). CONCLUSIONS: Our findings suggest that epigenetic silencing of RASSF1A gene expression by promoter hypermethylation could play an important role in primary esophageal squamous cell carcinogenesis.