GRHL3和c-Myc在食管鳞状细胞癌组织中的表达及其临床意义

目的:研究食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)患者肿瘤组织及癌旁组织中GRHL3和c-Myc的表达情况及其临床意义.方法:收集2015年4月至2016年7月在河北医科大学第四医院胸外科行肿瘤切除并经病理证实的64例ESCC患者的肿瘤组织、癌旁组织,采用Real-time PCR法和免疫组化法检测GRHL3和c-Myc基因的mRNA和蛋白表达情况,分析其与患者临床特征的关系.结果:与癌旁组织相比,ESCC组织中GRHL3 mRNA表达水平和蛋白阳性表达水平均显著升高[(2.85±2.83) vs(2.06±2.02),P＜0.01;81.30％ vs 25.00％,P＜0.01],ESCC组织中c-Myc mRNA表达水平和蛋白阳性表达水平均显著升高[5.13±5.11)vs (2.03±2.00),P＜0.01;42.20％ vs.20.30％,P＜0.01].ESCC组织中GRHL3 mRNA的表达与c-Myc mRNA的表达呈显著正相关关系(P＜0.05),GRHL3蛋白表达和c-Myc蛋白表达也呈显著正相关(P＜0.01).GRHL3蛋白表达和c-Myc蛋白表达与患者肿瘤浸润程度、淋巴结转移、临床分期、分化程度相关.结论:ESCC患者肿瘤组织中GRHL3与c-Myc表达水平显著提高,两者表达呈正相关,且两者与患者临床病理特征密切相关,可能是影响ESCC病理进程的重要因素.     

食管鳞状细胞癌组织Bin1基因启动子甲基化状态及其临床意义

目的:探讨食管鳞状细胞癌(esophageal squamous cell cancer,ESCC)患者的食管鳞癌组织、癌旁组织中Bin1基因启动子甲基化状态及其mRNA的表达及其临床意义.方法:采用实时荧光定量PCR(qRT-PCR)分别检测58例经病理证实的ESCC患者的食管鳞癌组织、癌旁组织中Bin1基因mRNA的表达情况;用甲基化特异性PCR(MSP)检测上述食管鳞癌组织中Bin1基因启动子甲基化状态,比较ESCC患者Bin1甲基化状态与临床病理分期的关系.结果:ESCC组织中Bin1基因启动子甲基化率明显高于癌旁组织(58.62％ vs 25.86％,x2=12.76,P＜0.01),Bin1甲基化状态与患者TNM分期、肿瘤侵润深度、分化程度、淋巴结转移相关(均P ＜0.05).ESCC组织中Bin1 mRNA的表达水平明显低于癌旁组织[(0.78 ±0.05) vs (1.03±0.03),t=9.643,P＜0.01)];发生Bin1甲基化的组织中Bin1 mRNA表达水平明显低于未发生甲基化的组织[(0.68±0.04) vs (0.85±0.07),t=2.476,P＜0.05].结论:Bin1基因启动子区甲基化状态可能与ESCC的发生密切相关,它是ESCC中Bin1 mRNA低表达或缺失的机制之一,且与ESCC进展和淋巴结转移有关.     

HHIP基因CpG岛高甲基化水平参与胃癌的发生

目的：观察人胃癌、癌旁组织与胃癌AGS细胞中人音猬因子相互作用蛋白（human hedgehog interacting protein,HHIP）基因启动子区域CpG岛的甲基化水平,探索其与胃癌发生的关系。方法：RT-PCR检测30例人胃癌组织、癌旁组织及AGS细胞中HHIP mRNA的表达,免疫组织化学方法和甲基化特异性PCR（methylation specific PCR,MSP）分别检测胃癌组织和癌旁组织的HHIP表达和HHIP基因启动子区域甲基化状态。AGS细胞予甲基化转移酶抑制剂5-氮杂-2′-脱氧胞苷（5-Aza-2′-deoxycitydine,5-Aza-dc）处理前后,RT-PCR、MSP和硫化测序PCR（bisulfite sequencing PCR,BSP）分别检测AGS细胞中HHIP mRNA表达、启动子区域甲基化水平变化、CpG岛甲基化位点数量的变化;分析HHIP基因启动子区CpG岛甲基化水平变化与HHIP mRNA表达水平变化之间的相关性。结果：胃癌组织中的HHIP mRNA（0.82± 0.38 vs 1.60±0.26,P=0.000）和蛋白（0.51±0.03 vs 0.83±0.27,P<0.05）的表达均低于癌旁组织,并且与年龄、性别、TNM分期、分化程度、淋巴结转移均无显著相关性（均P>0.05）。癌旁组织中HHIP基因启动子区甲基化水平显著低于胃癌和AGS细胞\[（17.7±3.59）% vs （62.9±614）%、（99.7±0.67）%,均P<0.05\]。AGS细胞在5-Aza-dc干预后HHIP mRNA表达明显增高（4.68±022 vs 0.21±012,P<0.01）,HHIP基因启动子区甲基化水平明显下降\[（10.1±0.21）% vs （90.2±0.67）%,P<0.01\], CpG岛甲基化位点明显减少,并且HHIP基因启动子区甲基化水平与mRNA表达呈负相关（r=-0.693,P=0.00）。结论：HHIP基因启动子区CpG岛的高甲基化水平可能通过抑制HHIP基因表达参与胃癌的发生。     

IL-6与VEGF在食管鳞状细胞癌中的表达及其临床意义

目的：检测食管鳞状细胞癌（esophageal squamous cell cancer,ESCC）患者血清中白细胞介素-6（interleukin-6,IL-6）和ESCC组织中血管内皮生长因子（vascular endothelial growth factor,VEGF）的表达及其两者关系,并研究两者表达对ESCC的临床意义。方法：收集河北医科大学第四医院胸外科于2014年1月至2015年1月期间行ESCC切除术的患者52例,每例患者均取原发灶组织和癌旁组织标本;同时在术前抽取患者外周血5 ml,再取52例健康体检者外周血5 ml为血清对照。应用ELISA法测定ESCC患者血清中IL-6的水平,免疫组化技术检测VEGF在ESCC组织中的表达,RT-PCR法检测肿瘤组织中IL-6和VEGF mRNA的表达情况。结果：ESCC患者血清IL-6表达水平为（116.71±25.98）pg/ml,明显高于健康对照组的\[（78.43±9.36）pg/ml\]（P＜0.05）,血清中IL-6的表达水平与患者肿瘤分化程度、TNM分期、肿瘤浸润深度和淋巴结转移相关（P＜0.05）。肿瘤组织VEGF蛋白阳性表达率明显高于癌旁组织（67.31% vs 32.69%,P<0.01),且与患者肿瘤分化程度、TNM分期、肿瘤浸润深度和淋巴结转移相关（P＜0.05）。肿瘤组织中IL-6 mRNA的表达与VEGF mRNA的表达呈显著正相关（r=7.113,P＜0.05）。结论：ESCC患者肿瘤组织中IL-6和VEGF均呈高表达且两者呈正相关,两者可能在ESCC侵袭和转移中发挥重要作用。     

食管鳞状细胞癌中ADAMTS1mRNA及其蛋白表达

摘 要：［目的］ 评价含凝血酶敏感素基序的去整合素金属蛋白酶1（a disintegrin and metalloproteinase with thrombospondin motifs 1,ADAMTS1）mRNA及其蛋白表达在食管鳞状细胞癌（esophageal squamous cell carcinoma,ESCC）发生发展中的意义。［方法］ 应用RT-PCR方法及免疫组织化学SP方法分别检测ESCC组织及癌旁正常组织中ADAMTS1 mRNA及其蛋白表达的情况。［结果］ ①ADAMTS1mRNA在ESCC组织中表达量显著性低于癌旁正常组织（0.394±0.123 vs 0.895±0.276,P<0.01）;ADAMTS1mRNA在无淋巴结转移组中的表达量显著性低于淋巴结转移组（0.298±0.102 vs 0.482±0.157,P<0.05）。②ESCC组织中ADAMTS1蛋白的阳性表达率显著性低于癌旁组织（38.9% vs 94.4%,P<0.01）。［结论］ ADAMTS1异常表达可能与ESCC的发生、发展及淋巴结转移密切相关。     

Atrogin-1基因在贲门腺癌组织中的表达及其异常甲基化

目的：探讨atrogin-1基因在贲门腺癌（gastric cardia adenocarcinoma,GCA）中的异常甲基化及表达,并分析其临床意义。方法: 选用河北医科大学第四医院2004—2008年间的贲门腺癌患者手术标本共139例,分别应用亚硫酸氢盐转换-甲基化特异性PCR（bisulfite conversion-methylation specific polymerase chain reaction,BS-MSP）、RT-PCR和免疫组织化学法检测贲门腺癌组织及相应癌旁（距癌灶边缘3～5 cm）组织中atrogin-1基因的甲基化、mRNA和蛋白表达情况,应用免疫组织化学法检测相应组织中Smad4蛋白的表达。结果: 贲门腺癌组织中atrogin-1基因启动子区的甲基化率\[44.6%（62/139）\] 显著高于癌旁组织\[3.6%(5/139)\]（χ2=63.891,P=0.001）,且atrogin-1基因的甲基化与TNM分期及肿瘤的组织学分化程度密切相关（χ2=6.144, P<0.05）。贲门腺癌组织中atrogin-1基因的mRNA和蛋白表达水平显著低于癌旁组织\[(0.482 5±0.175 4) vs (0.896 9±0.290 1),t=10.62, P=0.01;34.5% vs 82.0%, χ2=4.441,P=0.001\],且与其启动子区的甲基化状态之间有明显的相关性(r=-0.256,P=0.001)。贲门腺癌组织中Smad4蛋白表达的阳性率显著低于癌旁组织（46.0% vs 95.7%; χ2=2.945,P=0.001）,且与atrogin-1蛋白表达之间呈明显的正相关(r=0.604,P=0.001）。结论: Atrogin-1基因启动子区高甲基化导致的基因沉默可能是贲门癌组织中此基因表达降低的机制之一。     

IGFBP3基因在食管鳞状细胞癌中的表达及甲基化状态

目的：检测食管鳞状细胞癌(esophageal squamous cell cancer, ESCC)中胰岛素样生长因子结合蛋白3(insulin-like growth factor binding protein 3, IGFBP3)基因的表达情况及甲基化状态,探讨其与ESCC发生发展的关系。方法：收集河北医科大学第四医院2008至2011年间的82例ESCC手术患者的ESCC原发灶组织及癌旁正常黏膜组织。RT-PCR及甲基化特异性-PCR(methylation specific-PCR, MSP)的方法分别检测DNA甲基转移酶抑制剂5-氮杂-2′-脱氧胞苷(5-aza-2′-deoxycitydine, 5-Aza-dC)处理前后的ESCC细胞系(TE1、TE13、YES-2、T.TN、Eca109)及82例ESCC及相应癌旁组织中 IGFBP3 基因mRNA表达水平及甲基化状态,应用免疫组织化学方法检测IGFBP3在ESCC组织中的蛋白表达情况,并分析 IGFBP3 基因甲基化状态与其表达水平之间的关系。 结果： 在ESCC细胞株TE1、TE13、YES-2、T.TN、Eca109中, IGFBP3 基因mRNA均呈阴性或弱阳性表达,用5-Aza-dC培养处理后,其mRNA表达水平均呈现不同程度的增高（P＜0.05）;MSP检测结果显示,在ESCC细胞株TE1、TE13、T.Tn、Yes-2中 IGFBP3基因均呈高甲基化状态。在ESCC组织中 IGFBP3 mRNA表达显著低于癌旁组织\[（0.15±0.07）vs（0.88±0.32）,P＜0.01\],且IGFBP3蛋白在癌组织中的表达阳性率显著低于癌旁组织\[29.3%（24/82）vs 84.1%（69/82）,P＜0.01\],并与TNM分期密切相关（P＜0.05）;IGFBP3基因在ESCC组织中的甲基率为68.3%（56/82）,明显高于癌旁组织的15.9%（13/82）（P＜0.01）;IGFBP3基因在Ⅲ和Ⅳ期肿瘤组织中的甲基化率明显高于Ⅰ和Ⅱ期肿瘤组织（P＜0.05）,而该基因的甲基化率与肿瘤患者的组织学分级无相关性（P> 0.05）。IGFBP3基因甲基化状态与其表达之间有明显的相关性(P＜0.05)。结论： ESCC组织及细胞株中IGFBP3基因呈高甲基化状态,该基因的甲基化可能导致其表达下调,并有可能是ESCC的发生机制之一。     

B细胞易位基因2在食管鳞癌组织中的表达及临床意义

目的:探讨B细胞易位基因2（BTG2）在食管鳞状细胞癌（ESCC）组织中的表达及临床意义。方法:分析癌症基因组图谱（TCGA）数据库中BTG2 mRNA在ESCC组织及癌旁组织中的表达,受试者工作特征（ROC）曲线分析BTG2 mRNA表达量在预测ESCC患者疾病状态和放疗敏感性中的诊断价值;免疫组织化学检测我院184例ESCC患者癌组织标本BTG2蛋白表达,其中包括55例行根治性放疗术的ESCC患者,50例正常黏膜组织作为对照。分析BTG2蛋白表达情况与ESCC临床特征的关系。结果:与对照组比较,BTG2 mRNA在ESCC组织中表达下降（5.08±1.06 vs 5.91±1.29,t=2.387,P=0.019）,与放疗敏感组患者比较,BTG2 mRNA在放疗抵抗的ESCC患者癌组织中表达下降（3.82±0.97 vs 5.44±0.73,t=4.935,P＜0.001）,ROC结果显示BTG2 mRNA在鉴别放疗敏感与放疗抵抗患者时特异性为95.65%,敏感性为75%（AUC=0.902,P＜0.001）;免疫组织化学结果显示ESCC组织中BTG2阳性表达（103/184）低于正常黏膜组织（42/50）,差异有统计学意义（χ2=13.10,P＜0.001）;BTG2表达在放疗敏感和放疗抵抗组织中的阳性表达率分别为57.9%（22/38）和23.5%（4/17）,差异有统计学意义（χ2=5.565,P=0.018）;ESCC患者BTG2蛋白表达差异在不同性别、年龄、T分期及M分期中无统计学差异（P＞0.05）,与N分期（χ2=4.134,P=0.042）及临床分期（χ2=5.303,P=0.021）相关;单因素分析结果显示,肿瘤分级（HR=0.500,95%CI:0.317～0.790,P=0.003）、N分期（HR=0.275,95%CI:0.157～0.479,P=0.000）、M分期（HR=0.317,95%CI:0.151～0.665,P=0.002）、临床分期（HR=0.269,95%CI:0.167～0.434,P=0.000）及BTG2表达（HR=1.956,95%CI:1.242～3.079,P=0.003）与ESCC患者总生存（OS）有关;多因素分析结果显示,肿瘤分级（HR=0.613,95%CI:0.381～0.987,P=0.044）、N分期（HR=0.507,95%CI:0.259～0.991,P=0.047）、临床分期（HR=0.504,95%CI:0.278～0.916,P=0.025）及BTG2表达（HR=1.608,95%CI:1.011～2.558,P=0.045）影响ESCC患者的OS。结论:BTG2具有作为预测ESCC进展及放疗敏感性生物标记物的潜在价值,并且是影响ESCC患者总生存率的独立预后因素。     

Ras相关区域家族7基因在食管鳞状细胞癌中的表达及其临床意义

目的：检测人食管鳞癌 (esophageal squamous cell carcinoma, ESCC)组织中Ras相关区域家族7（Ras-association domain family 7,RASSF7）基因的mRNA、蛋白表达情况及其甲基化状态,探究RASSF7 在ESCC发生发展中的作用。方法：组织标本取自河北医科大学第四医院2011—2012年间手术切除的69例ESCC原发灶组织及癌旁组织。分别应用RT-PCR及甲基化特异性PCR(methylation specific polymerase chain reaction,MSP)方法检测DNA甲基转移酶抑制剂5-氮杂-2’-脱氧胞苷（5-aza-2’-deoxycitydine, 5-Aza-dC）处理前后的4株食管癌细胞系（TE13、T.Tn、YES-2、Ec109）和69例病灶组织及其癌旁组织中RASSF7 mRNA表达水平及甲基化状态,应用免疫组织化学方法检测69例ESCC组织及相应癌旁组织中RASSF7的蛋白表达。结果：RASSF7基因在TE13、T.Tn、YES-2细胞系中表达阳性,在Ec109细胞系中表达缺失;经5-Aza-dC处理后,RASSF7在TE13、T.Tn、YES-2细胞中表达下调,在Ec109细胞中表达阳性。5-Aza-dC处理前后4株食管癌细胞系中均未检测到RASSF7 的甲基化。人ESCC组织中RASSF7 的mRNA相对表达量（0.63±0.08 vs 0.42±0.20,P<0.01）与蛋白表达阳性率\[81.2%（56/69） vs 53.6%（37/69）,P<0.01\]均显著高于相应癌旁组织,且均与患者的淋巴结转移情况及分化程度有关（P<0.05或P<001）,与TNM分期、年龄和性别无关（均P>0.05）。ESCC组织和相应癌旁组织中均未检测到RASSF7的甲基化。结论：4株食管癌细胞系、人ESCC组织和癌旁组织中RASSF7基因的表达差异与RASSF7 本身甲基化状态无关,ESCC组织中RASSF7 的高表达可能参与了ESCC的发生及转移。     

转录因子 SOX7 基因在贲门腺癌中的异常表达及其甲基化状态

目的：检测贲门腺癌（gastric cardia adenocarcinoma,GCA）组织及相应癌旁非肿瘤组织中Y性别决定区基因7（sex determining region Y-box 7, SOX7 ）的甲基化状态及其对 SOX7 mRNA表达、Wnt通路中心因子β-catenin异质表达的影响及与临床病理特征之间的相关性。方法：选择河北医科大学第四医院胸外科2006-2012年手术切除的GCA及癌旁组织各130例,应用甲基化特异性PCR（methylation specific PCR, MSP）、RT-PCR分别检测130例GCA及癌旁组织中 SOX7 基因的甲基化状态及其mRNA表达情况,应用免疫组织化学方法检测标本中β-catenin蛋白的表达。分析 SOX7 的甲基化状态与临床病理特征、 SOX7 mRNA表达、β-catenin蛋白的异质表达及上消化道肿瘤家族史（upper gastrointestinal cancers,UGIC）的关系。 结果 ：GCA组织中 SOX7 的甲基化率显著高于癌旁组织为\[57.7%（75/130） vs 30.8%（40/130）, P <0.01\],其高甲基化仅与肿瘤患者的淋巴结转移情况有关（ P <0.05）,与肿瘤组织的病理分级及临床分期均无关（ P >0.05）。GCA组织中 SOX7 mRNA的表达量明显低于癌旁非肿瘤组织\[(0.414±0.054) vs (0.695±0.034), P <0.01]\],其β-catenin蛋白的异质表达率显著高于癌旁组织（85.4% vs 43.1%, P <0.01）;GCA组织中 SOX7 基因mRNA表达情况及β-catenin蛋白的异质表达率均与该基因的甲基化状态有关（ P <0.05）,并且 SOX7 基因的甲基化状态与贲门癌患者的上消化道家族史密切相关（ P <0.01）。结论： CpG岛甲基化是 SOX7 基因表达下调的机制之一,并可能通过Wnt/β-catenin信号转导通路的激活在贲门腺癌的发生中扮演着重要的角色。     

Bacteria and cancer--antagonisms and benefits

There is considerable historical and recent evidence concerning the antagonisms between acute bacterial infections or their toxins and cancer and allied diseases. These data provide renewed incentives to undertake clinical programmes with mixed bacterial vaccines in many countries at the present time.     

Religiosity and physical and emotional functioning among African American and White colorectal and lung cancer patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

Genetic polymorphisms of alcohol and aldehyde dehydrogenases and risk for esophageal and head and neck cancers

Alcoholic beverages are causally related to cancer of the oral cavity, pharynx, larynx and esophagus. Ethanol is oxidized to acetaldehyde and then to acetate by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), both of which have genetic polymorphisms. A review of case-control studies of the effects of ALDH2, ADH2 and ADH3 genotypes shows consistently positive associations between inactive heterozygous ALDH2 and the less-active ADH2 genotypes and the risk for esophageal cancer in East Asian heavy drinkers and this enzyme-related vulnerability may extend to light-to-moderate drinkers. Some studies suggest similar associations with the risk for head and neck cancer in moderate-to-heavy-drinking Japanese. An established carcinogen in experimental animals, acetaldehyde can interact with human DNA. ALDH2-associated cancer susceptibility fits into a scenario in which acetaldehyde plays a critical role in the development of human cancer. Alcohol flushing and drinking behavior may partly explain this carcinogenic effect in carriers of less-active ADH2 genotypes. Whether the ADH3 genotype influences head and neck cancer risk in Western nations is controversial. Professional and public education about risky conditions connected to the ALDH2 and ADH2 genotypes and environmental factors is important in a new strategic approach to the prevention of alcohol-related cancers in East Asians. The use of simple tests to identify inactive ALDH2 on the basis of alcohol flushing responses could benefit many people, by helping them to identify their own cancer risks. Such testing could also help clinicians diagnose esophageal cancer earlier, through the use of endoscopic screening in the high-risk population.     

Religiosity and Physical and Emotional Functioning Among African American and White Colorectal and Lung Cancer Patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

Renal irradiation and the pharmacology and toxicity of methotrexate and cisplatinum

We used a rat model to study the effects of renal irradiation on the pharmacology of methotrexate (MTX) and cisplatinum (cis-Pt). Unanesthetized rats were given bilateral kidney irradiation (20 Gy in 9 fractions). At 9 months after irradiation, 3% of the animals had died and survivors showed moderately impaired renal function. At 15 months, 30% of the animals had died and survivors showed severely impaired renal function. Some animals were given i.v. MTX 1 week to 15 months after irradiation. In irradiated rats, the area under the MTX plasma clearance curve equaled that of controls through 6 months, and was significantly above controls from 9 months on. Other animals were given i.p. cis-Pt 1 week to 9 months after irradiation. The acute toxicity of cis-Pt was the same in control and irradiated rats when cis-Pt was given immediately before or after irradiation. Beginning 3 months after irradiation there was a progressive increase in cis-Pt toxicity and a simultaneous decrease in urinary platinum excretion. Irradiated animals that survived cis-Pt treatment showed increased radiation nephritis; the greatest effect occurred when cis-Pt was given 3 months or more after irradiation. MTX and cis-Pt clearance decreased when renal dysfunction was first observed and changes in renal function preceded changes in drug clearance and toxicity.