胃癌组织中E—cadherin基因启动子异常甲基化状态观察

目的观察胃癌组织中E—cadherin基因启动子异常甲基化状态。方法采用甲基化特异性聚合酶链反应（MSP）技术检测54例胃癌及其癌旁正常组织中的E—cadherin基因启动子异常甲基化状态,并分析E-cadherin基因启动子异常甲基化状态与胃癌临床病理参数的关系。结果胃癌组织中的E-cadherin基因启动子异常甲基化发生频率为48．10％（26／54）,明显高于癌旁正常组织中的11．11％（6／54）。E—cadherin基因启动子异常甲基化频率与胃癌分化程度、病理类型、浸润深度以及淋巴结转移、临床分期有关（P均〈0．05）。结论胃癌组织中E-cadherin基因启动子异常甲基化频率较高,并与胃癌浸润、转移有关。     

No association of the matrix metalloproteinase 1 promoter polymorphism with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in northern China

AIM: To investigate association of the 2Gor1Gsingle nudeotide polymorphism (SNP) in matrix metalloproteinase 1 (MMP1) promoter with susceptibility to esophageal squam-ous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of North China. METHODS: MMP1 promoter SNP was genotyped by polymerase-chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis in 417 cancer patients (234 ESCC and 183 GCA) and 350 healthy controls. RESULTS: The genotype frequencies of the MMP1 promoter SNP in healthy controls were 55.4% (2G/2G), 30% (1G/2G) and 14.6% (1G/1G), respectively. The genotype and allelotype distribution in ESCC and GCA patients was not significantly different from that in healthy controls (all lvalues were above 0.05). Compared with the 1G/1Ggenotype, neither the 2G/2Gnor in combination with the 1G/2G genotype significantly modified the risk of developing ESCC and GCA, the adjusted odds ratio was 1.28 (95%CI = 0.78-2.09), 1.23 (95%CI = 0.38-2.05) in ESCC and 1.39 (95%CI = 0.80-2.41), 1.34 (95%CI = 0.74-2.40) in GCA, respectively. When stratified by smoking status and family history of upper gastrointestinal cancer, the 2G/2Ggenotype alone or in combination with the 1G/2G genotype also did not show any significant influence on the risk of ESCC and GCA development. In addition, influence of the MMP1 SNP on lymphatic metastasis in ESCC and GCA was also not obs-erved. CONCLUSION: The 2Gor 1GSNP in the MMP1 promoter might not modify the risk of ESCC and GCA development and might not be used as a stratification marker to predict the potential of lymphatic metastasis in these two tumor types.     

Aberrant p16 methylation, a possible epigenetic risk factor in familial esophageal squamous cell carcinoma

Aim. Detection of methylation in the p16 gene, an inhibitor of cyclin D-dependent protein kinase, as a new tumor marker for early detection of esophageal squamous cell carcinoma (ESCC) in DNA derived from blood and serum. Method. A large family with clustering of ESCC was assessed in Khorasan province in northeastern Iran. The family had three histologically proven cases of ESCC in two consecutive generations and several other deceased cases with histories of ESCC. DNA from blood of 28 living family members in three consecutive generations, 30 sporadic ESCC cases (from serum, blood, and tumor tissues), and 30 healthy volunteers (from blood) were examined for the methylation status of p16 promoter using methylation-specific PCR (MSP). Results. Aberrant p16 promoter methylation was found in 64.3% (n=28) of ESCC family members and none (n=30) of our normal volunteers. Five of the 28 family members with esophageal cancer symptoms had negative endoscopy results for ESCC, while four of these members had p16 hypermethylation in their blood. The family members with negative endoscopy and positive p16 promoter methylation are being monitored closely for signs of ESCC development through regular check-ups and chromoendoscopies. In sporadic ESCC in northeastern Iran, 73.3% (n=30) of tumor tissue samples had p16 hypermethylation. Serum and blood samples from the same patients showed p16 hypermethylation in 26.6% and 43.3% of the samples, respectively. Conclusion. Aberrant p16 methylation may be a valuable diagnostic tool as a tumor marker for the early identification of individuals in high risk ESCC families.     

5-Aza-CdR对食管鳞癌甲基化基因的影响

目的探讨5-aza-CdR对食管鳞癌甲基化基因的影响。方法应用人类全基因组寡核苷酸微阵列芯片,荧光双交换法检测5-aza-CdR干预的食管鳞癌细胞Eca-109与正常培养的Eca-109细胞中的差异表达基因,并进行生物信息学分析。结果经统计学分析,共筛选获得384个差异表达基因,其中303个基因表达上调,81个基因表达下调;差异基因的功能涉及信号转导、物质合成代谢、细胞周期、细胞增殖、细胞凋亡、DNA转录、DNA复制、DNA修复、氧化还原、物质运输、免疫反应等;上调表达基因中有138个基因分别含有1～5个CpG岛序列,占总上调基因的45.54%。结论5-aza-CdR可以影响食管鳞癌细胞中基因异常的甲基化修饰,调控肿瘤细胞的异常凋亡和分化,为进一步研究食管鳞癌致病分子机制,提供表观遗传学的依据。     

Aberrant p16 methylation, a possible epigenetic risk factor in familial esophageal squamous cell carcinoma

AIM: Detection of methylation in the p16 gene, an inhibitor of cyclin D-dependent protein kinase, as a new tumor marker for early detection of esophageal squamous cell carcinoma (ESCC) in DNA derived from blood and serum. METHOD: A large family with clustering of ESCC was assessed in Khorasan province in northeastern Iran. The family had three histologically proven cases of ESCC in two consecutive generations and several other deceased cases with histories of ESCC. DNA from blood of 28 living family members in three consecutive generations, 30 sporadic ESCC cases (from serum, blood, and tumor tissues), and 30 healthy volunteers (from blood) were examined for the methylation status of p16 promoter using methylation-specific PCR (MSP). RESULTS: Aberrant p16 promoter methylation was found in 64.3% (n = 28) of ESCC family members and none (n = 30) of our normal volunteers. Five of the 28 family members with esophageal cancer symptoms had negative endoscopy results for ESCC, while four of these members had p16 hypermethylation in their blood. The family members with negative endoscopy and positive p16 promoter methylation are being monitored closely for signs of ESCC development through regular check-ups and chromoendoscopies. In sporadic ESCC in northeastern Iran, 73.3% (n = 30) of tumor tissue samples had p16 hypermethylation. Serum and blood samples from the same patients showed p16 hypermethylation in 26.6% and 43.3% of the samples, respectively. CONCLUSION: Aberrant p16 methylation may be a valuable diagnostic tool as a tumor marker for the early identification of individuals in high risk ESCC families.     

Prognostic and clinical impact of sarcopenia in esophageal squamous cell carcinoma

Recently, depletion of skeletal muscle mass (sarcopenia) has been linked to poor prognosis in several types of cancers, but has not been investigated in esophageal squamous cell carcinoma (ESCC). This retrospective study investigates the relationship between sarcopenia and clinical outcome in ESCC patients treated by surgical resection or definitive chemoradiation therapy (dCRT). The study was retrospectively conducted in a single academic hospital in Kumamoto, Japan, and involved 325 ESCC patients (256 surgical cases and 69 dCRT cases) treated between April 2005 and April 2011. Skeletal muscle mass was quantified by radiologic measures using standard computed tomography scans. The skeletal muscle tissue in the 325 ESCC patients was distributed as follows: mean: 47.10; median: 46.88; standard deviation (SD): 7.39; range: 31.48–71.11; interquartile range, 46.29–47.90. Skeletal muscle tissue was greater in male patients than in female patients (P < 0.0001), but was independent of other clinical and tumor features. Sarcopenia was not significantly associated with overall survival (log rank P = 0.54). Lymph node involvement significantly altered the relationship between sarcopenia and survival rate (P for interaction = 0.026). Sarcopenia significantly reduced the overall survival of patients without lymph node involvement (log rank P = 0.035), but was uncorrelated with overall survival in patients with lymph involvement (log rank, P = 0.31). The anastomosis leakage rate was significantly higher in the sarcopenia group than in the non‐sarcopenia group (P = 0.032), but other surgical complications did not significantly differ between the two groups. Sarcopenia in ESCC patients without lymph node involvement is associated with poor prognosis, indicating sarcopenia as a potential biomarker for identifying patients likely to experience an inferior outcome. Moreover, sarcopenia was associated with anastomosis leakage but no other short‐term surgical outcome.     

Amplification of the telomerase RNA component gene as a new genetic marker for disease progression and prognosis in esophageal squamous cell carcinoma

Amplification of the human telomerase RNA component (TERC) gene was found in esophageal squamous cell carcinoma (ESCC). However, its roles in the progression and prognosis of ESCC have not been well understood. The amplification of TERC in normal mucosa, low‐grade and high‐grade intraepithelial neoplasia, and invasive ESCC samples were evaluated using a fluorescence in situ hybridization assay. The amplification of TERC invariably occurred in high‐grade intraepithelial neoplasia and invasive ESCC, partially occurred in low‐grade intraepithelial neoplasia specimens, and seldom occurred in normal mucosa. The average signal ratio of TERC to chromosome 3 centromere‐specific probe (TERC/CSP3) was 1.00 ± 0.01 (average ± standard deviation) in normal mucosas, 1.01 ± 0.08 in low‐grade intraepithelial neoplasias, 1.39 ± 0.26 in high‐grade intraepithelial neoplasias, and 1.56 ± 0.41 in invasive ESCC. High TERC/CSP3 ratio was positively associated with lymph node metastasis (P = 0.005) and advanced tumor stage (P = 0.045). Patients with high amplification of TERC had poor survival (P = 0.01). The amplification of TERC could be used as a new genomic marker for disease progression and prognosis of ESCC. The amplified TERC gene may be a potential therapeutic target for ESCC.     

RASSF1A基因启动子区甲基化在贲门腺癌、食管下段鳞癌组织中的变化及临床意义

目的:比较RASSF1A基因甲基化在贲门腺癌、食管下段鳞癌不同病理阶段组织中的共性和差异.方法:33例贲门腺癌和36例食管下段鳞癌手术切除标本纳入本研究,每例标本选取癌组织,癌旁不典型增生组织(距癌3-5 cm),癌旁正常组织(距癌>5 cm)组织各1块,采用甲基化特异性PCR检测RASSF1A基因启动子区的甲基化情况...     

Upregulated ataxia‐telangiectasia group D complementing gene correlates with poor prognosis in patients with esophageal squamous cell carcinoma

The ataxia‐telangiectasia group D complementing gene (ATDC) plays significant roles in various human cancers. However, the clinical significance of ATDC in esophageal squamous cell carcinoma (ESCC) has not been investigated. The ATDC messenger RNA level of 40 paired ESCC and nonneoplastic tissues were evaluated using quantitative real‐time polymerase chain reaction, 10 pairs of which were also used for Western blot analysis. In addition, immunohistochemical staining was performed to detect the ATDC expression in 118 paraffin‐embedded cancerous and matched nonneoplastic tissues, and the correlation of ATDC expression with the clinicopathological parameters and prognosis of the ESCC patients was analyzed. The quantitative real‐time polymerase chain reaction, immunohistochemical staining, and Western blot results demonstrated that the expression level of ATDC was significantly higher in ESCC tissue than in matched noncancerous tissues. Both ATDC messenger RNA and protein expression in the ESCC tissue were significantly correlated with tumor differentiation, stage, and lymph node metastasis. However, there was no significant difference in ATDC expression based on patient age or gender. Moreover, the results of both univariate and multivariate analyses showed that increased ATDC expression was correlated with a shorter 5‐year survival time for ESCC patients after surgery. We concluded that increased ATDC expression is associated with poor clinical outcomes and that this marker might be a useful indicator for prognosis and a promising target for therapy in ESCC patients.     

Role of podoplanin expression in esophageal squamous cell carcinoma: a retrospective study

Lymphatic metastasis is the predominant cause of the low overall survival of patients with esophageal squamous cell carcinoma (ESCC), as there are no faithful methods available predicting early metastasis. Recent studies suggest an effect of podoplanin expression on metastatic spreading to lymph nodes. The purpose of this study was to investigate the influence of podoplanin expression on lymphatic metastasis and tumor cells, and to find the relationship between podoplanin expression and prognosis of patients with ESCC. We evaluated the level of podoplanin expression on tumor cells and the lymphatic vessel density change of tumor mass compared with normal tissue from the same patient through D2-40 immunohistochemistry staining, and analyzed associations between these two variables and various clinicopathologic parameters individually or conjunctively. There was an association between podoplanin expression and the frequency of lymph node metastases. In 45 patients (80%), podoplanin was expressed on the tumor cells. Twenty-one patients (37.5%) showed high levels of expression. The 5-year disease-free survival rate (5%) for patients with high levels of podoplanin expression was significantly lower (P < 0.001) than for patients with low and moderate expression of podoplanin (54% and 27%, respectively). We concluded that podoplanin is expressed frequently in ESCC, and that the expression of podoplanin on cancer cells, lymphatics, or both is correlated with lymphatic metastasis and clinical outcome.     

Association of mu‐opioid receptor expression with lymph node metastasis in esophageal squamous cell carcinoma

The mu‐opioid receptor (MOR), a membrane‐bound G protein‐coupled receptor, is the main target for opioids in the nervous system. MOR1 has been found in several types of cancer cells and reported to be involved in tumor progression and metastasis. However, the expression and clinical significance of MOR1 in esophageal squamous cell carcinoma (ESCC) remain unclear. In our study, the expression of MOR1 was confirmed in ESCC cell lines (KYSE180, KYSE150, and EC109) by Western blot. MOR1 was also detected on tissue microarrays of ESCC samples in 239 cases using immunohistochemical staining. We found that MOR1 was mainly located in the cytoplasm and occasionally occurred in the membrane or nucleus of ESCC cells. Moreover, results indicated that MOR1 expression in the cytoplasm was associated with lymph node metastasis (R = 0.164, P = 0.008, Kendall's tau‐b‐test). No more associations were found between MOR1 expression status and other clinical parameters. However, no statistical significant differences were found between MOR1 expression in the cytoplasm, nucleus/membrane, and the overall survival of ESCC patients (P = 0.848; P = 0.167; P = 0.428, respectively, log‐rank test). Our results suggest that the cytoplasmic MOR1 may be a high‐risk factor for lymph node metastasis of ESCC patients. We also hypothesize that MOR1 agonists used in ESCC patients should be prudent, and opioid receptor antagonists may be novel therapeutic drugs for ESCC patients.     

Demethylation-mediated upregulation of melanoma-associated antigen-A11 correlates with malignant progression of esophageal squamous cell carcinoma

BackgroundThe expression and methylation status of oncogenes are closely related to the onset and progression of cancer.AimsTo explore the role and methylation status of melanoma-associated antigen-A11 in the pathogenesis of esophageal squamous cell carcinoma.Methods116 esophageal squamous cell carcinoma patients with tumor tissues and corresponding adjacent normal tissues were obtained. The expression level and methylation status of melanoma-associated antigen-A11 in esophageal cancer cell lines and esophageal squamous cell carcinoma tissues were determined respectively.ResultsSignificant up-regulation of melanoma-associated antigen-A11 was detected in esophageal cancer cell lines and esophageal squamous cell carcinoma tissues. Up-regulation of melanoma-associated antigen-A11 contributed to proliferation and invasion in cancer cells. Hypomethylation of the CpG site was associated with pathological differentiation, clinical stage, tumor size, lymph node metastasis and distant metastasis. Esophageal squamous cell carcinoma patients in stage III and IV, with high expression of melanoma-associated antigen-A11 or hypomethylation of the CpG site within the promoter demonstrated poor survival.ConclusionMelanoma-associated antigen-A11 is up-regulated in esophageal squamous cell carcinoma at least partly by hypomethylation of the CpG site within the promoter and this hypomethylation may affect the prognosis of esophageal squamous cell carcinoma patients.     

Meta‐analysis: non‐steroidal anti‐inflammatory drug use and the risk of esophageal squamous cell carcinoma

The relationship between non‐steroidal anti‐inflammatory drug (NSAID) use and esophageal squamous cell carcinoma (ESCC) has remained unclear. To evaluate the relationship between NSAID use and risk of ESCC, we searched the MEDLINE, Biosis, Web of Science and ISI proceedings databases up to September 2010, together with a manual search of reference lists of relevant articles. Studies evaluating the association between exposure to NSAIDs and risk of ESCC were included. The analyses used random‐effect or fixed‐effect model based on homogeneity analysis. Seven studies (six case‐control studies and one nested case‐control study) were included in this meta‐analysis. NSAID use was associated with a reduced risk of ESCC (odds ratio = 0.58, 95% confidence interval = 0.47 to 0.72). Specific analysis for aspirin and non‐aspirin NSAIDs yielded similar results. There was a protective association between NSAIDs and ESCC. This finding warrants more prospective studies evaluating the relationship between NSAIDs and ESCC.