中国非小细胞肺癌患者血清DAPK基因、p16基因启动子甲基化的分析(英文)

目的分析65名中国南京军区南京总医院的非小细胞肺癌(NSCLC)住院患者血清中DAPK、p16启动子区域甲基化的改变状况及其临床意义。方法运用甲基化特异性PCR技术,检测65例NSCLC患者血清DAPK、p16基因启动子区域甲基化的改变情况,并分析与临床病理资料的关系。结果 NSCLC患者血清DAPK基因甲基化检出率为30．8％(20/65),p16基因甲基化检出率为43．1％(28/65),而正常对照组和良性肺部疾病组血清未检出DAPK基因、p16基因甲基化,DAPK基因、p16基因甲基化检出率与NSCLC病理类型、分期及转移状态无明显关系。结论 DAPK基因、p16基因检出启动子区域异常甲基化是NSCLC早期辅助诊断的分子标志物之一。     

胃癌中E-cadherin和DAPK基因启动子异常甲基化的研究

目的:检测胃癌中死亡相关蛋白激酶(death-associated protein kinase,DAPK)基因和上皮钙粘蛋白(epithelial cadherin,E-cadherin)基因启动子区CpG岛甲基化状态,并探讨两个基因甲基化改变的特点及其与临床病理特征、患者一般资料之间的关系.方法:采用目前常用的甲基化特异性PCR(Methylation-specific PCR.MSP)方法检测4l例胃癌组织和20例正常对照组织中DAPK、E-cadherin基因启动子区甲基化状态并进行统计分析.结果:41例胃癌组织中DAPK、E-cadherin基因启动子区甲基化阳性率分别为68.3%(28/41)和46.3%(19/41),20例正常时照组织中未检测到DAPK、E-cadherin基因启动子区发生甲基化,两个基因在胃癌组织中的甲基化率明显高于正常对照组织(P<0.05),但DAPK和E-cadherin基因启动子甲基化在胃癌的发生中无协同性(相关性和一致性).胃癌组织中一个基因发生甲基化的检出率为78.0%(32/41);胃癌组织中DAPK基因启动子区甲基化与淋巴结转移、分化程度相关(P<0.05),而E-cadherin基因启动子区甲基化则与淋巴结转移和浸润深度有相关性(P<0.05).两个基因启动子区异常甲基化与被检查者肿瘤的大小、肿瘤的部位等临床病理特征以及被检者的性别、年龄不具有相关性.结论:DAPK、E-eadherin基因启动子区甲基化是胃癌发生、发展过程中的频发事件,通过检测胃粘膜组织中两个基因启动子区甲基化状况,可能会对胃癌的早期诊断及判断预后提供一定的参考价值;联合检测两个基因甲基化状态优于各单个基因检测.     

肝癌患者血清p16、Runx3基因甲基化及其与临床病理参数的相关性

目的:探讨血清p16和Runx3基因启动子区CpG岛异常甲基化与原发性肝癌(hepatocellular carcino-ma,HCC)关系。方法:采用甲基化特异性PCR(MSP)法对56例HCC患者、40名健康体检者、40例乙肝患者和40例肝硬化患者血清p16和Runx3基因甲基化状况进行检测。结果:HCC患者血清p16和Runx3基因甲基化检出率分别为69.6%(39/56)和66.1%(37/56),而肝硬化患者、乙肝患者和健康体检者均未检出基因甲基化,与HCC组比较差异均有统计学意义(P<0.05)。Logistic回归分析结果均未发现以α=0.05为显著性水准进入回归模型的变量。结论:HCC患者血清中可检测到p16和Runx3基因的甲基化,有助于肝癌的诊断、治疗和预后评估。     

抑癌基因甲基化谱分析在肝细胞癌诊断中的价值

目的 探讨beclin 1、RASSFA-1、p16、DAPK等抑癌基因启动子异常甲基化及其联合检测在肝癌早期诊断中的价值。方法 采用甲基化特异性PCR（methylation specific PCR,MSP）法,检测37例肝癌组织和相应的癌旁组织中beclin 1、RASSFA-1、p16和DAPK 4 种基因启动子区甲基化状态。通过与AFP的单一ROC曲线分析及其逐步Logistic回归的ROC曲线下面积,对单一及组合方式的敏感度、特异性、Youden指数和阳性似然比/阴性似然比进行比较分析。结果 肝癌组织中beclin 1、RASSFA-1、p16和DAPK 基因启动子区甲基化检出率分别为5.4%（2/37）、94.6%（35/37）、 73.0%（27/37 ）和 35.1% （13/37）,其中相应的癌旁组织RASSFA-1、p16和DAPK基因启动子区甲基化检出率与肝癌组织比较差异有统计学意义（P<0.05）,RASSFA-1基因启动子区甲基化和AFP肿瘤标志物联合检测可显著提高肝癌检测的敏感度（95.0%）和特异性（97.3%）,曲线下面积最大（0.903）,并获得高于单一基因甲基化的敏感度、Youden指数、阳性似然比/阴性似然比。结论 RASSFA-1基因启动子区甲基化和AFP肿瘤标志物联合检测,可能是早期筛查和诊断肝细胞癌的有效指标。     

鼻咽癌组织中DAPK基因启动子甲基化的研究

目的:分析鼻咽癌组织中DAPK基因启动子的甲基化状态,探讨DAPK基因启动子甲基化与鼻咽癌的关系。方法:应用甲基化特异性PCR技术检测48例鼻咽癌组织、26例慢性鼻咽黏膜炎症组织的DAPK基因启动子甲基化状态,比较鼻咽癌和慢性鼻咽黏膜炎症组织的DAPK基因启动子甲基化率。结果:鼻咽癌组织的DAPK基因启动子甲基化率为75%,而慢性鼻咽黏膜炎症组织中未检测到DAPK基因启动子甲基化。结论:鼻咽癌组织中DAPK基因启动子存在高甲基化水平,检测DAPK基因启动子甲基化或许能为鼻咽癌的诊断提供依据。     

抑癌基因高甲基化与胃癌化疗药物敏感相关性的探讨

目的:研究抑癌基因p16、MG-MT、DAPK启动子甲基化与胃癌患者临床病理参数及对化疗药物敏感性的关系。方法:MSP检测38例胃癌组织及相应20例正常胃黏膜组织p16、MGMT、DAPK基因启动子甲基化状况。MTT法检测5-氟尿嘧啶(5-FU)、顺铂(DDP)、多柔比星(ADM)、吉西他滨(GEM)、丝裂霉素(MMC)等化疗药物单用及联用对38例原代胃癌细胞的抑制率。并比较p16、MGMT、DAPK甲基化与非甲基化组胃癌对化疗药物的敏感性差异。结果:三基因在胃癌组织中甲基化率均明显高于正常胃黏膜组织,P<0.05。p16甲基化与Lauren分型相关,DAPK甲基化与胃癌TNM分期相关,P值均<0.05。p16甲基化组5-FU、5-FU+DDP+ADM及5-FU+DDP+GEM抑制率高于非甲基化组;DAPK甲基化组MMC抑制率低于非甲基化组,P值均<0.05。结论:抑癌基因启动子高甲基化可能是胃癌发生发展过程中的重要事件。不同抑癌基因发生甲基化对胃癌化疗敏感性影响可能不同。     

肺癌血清p16基因启动子畸变甲基化检测

[目的]检测肺癌患者血清DNA中p16基因启动子畸变甲基化,并探讨其在肺癌早期诊断和预后评价中的作用。[方法]收集新鲜的肺癌组织及其对应的血清标本69例,肺良性疾病患者标本25例,健康人血清标本10例。采用PCR技术分别检测肿瘤组织DNA和血清游离DNA中p16基因启动子畸变甲基化的状况。[结果]28.99%(20/69)肺癌组织出现p16高甲基化;出现高甲基化的组织标本相对应的20例血清中有15例(75%)出现p16基因高甲基化。肺良性疾病患者及正常人的血清、DNA中均未见p16基因高甲基化。血清p16基因高甲基化与肺癌TNM分期及分化程度密切相关(P<0.01或P<0.05)。[结论]非小细胞肺癌血清中p16基因甲基化状况的检测,可能有助于非小细胞肺癌的早期诊断或预后评估。     

原发性肝癌(HCC)TACE治疗前后p16、AFP、sVEGF的动态变化

目的:探讨HCC患者经肝动脉灌注化疗栓塞术(TACE)治疗前、后p16基因甲基化、AFP和sVEGF变化的临床意义。方法:采用甲基化特异性PCR(MSP)法检测56例HCC患者血清p16基因甲基化率,ELISA法检测AFP和sVEGF。结果:HCC患者血清p16基因甲基化检出率为69.6%(39/56),AFP阳性检出率为80.4%(45/56),sVEGF阳性检出率为73.2%(41/56)。结论:血清DNA中p16基因甲基化检测可作为HCC分子诊断标志,三者联合检测对肝癌诊断有实用价值。     

肝癌患者血清p16甲基化检测

肝细胞癌(hepatocellular carcinoma，HCC)是原发性肝癌的主要病理类型，其发病的分子机制尚不清楚。近年来发现，p16基因5’-启动子区CpG岛异常甲基化在许多肿瘤组织及患者血清或血浆循环核酸(ciI℃ulatingnucleicacid，CNA)中有较高检出率。我们用甲基化特异性PCR(methvlationspecific PCR，MSP)检测HCC患者血清p16基因启动子区CpG岛甲基化状态，分析p16甲基化与HBV感染、AFP等的关系，并探讨其临床意义。     

宫颈癌组织中DAPK基因启动子甲基化的研究

背景与目的:已有研究表明DNA异常甲基化在肿瘤的发生、发展中发挥了重要作用.本研究旨在探讨宫颈癌组织中DAPK(death-associated protein kinase)启动子甲基化与基因失活的关系.方法:应用甲基化特异性PCR (methylation-specific PCR,MSP)技术检测52例宫颈癌、60例宫颈上皮内瘤样病变(cervical intraepithelial neoplasia,CIN)和20例正常宫颈鳞状上皮DAPK启动子甲基化状况,应用免疫组化方法检测其蛋白表达;分析DAPK启动子甲基化和基因失活与宫颈癌临床病理因素之间的关系.结果:正常宫颈组织不存在DAPK基因启动子CpG岛甲基化,而CIN和宫颈癌中的DAPK甲基化率分别为18.3%(11/60)、65.4%(34/52),三者之间的差异有统计学意义(P<0.05).宫颈鳞癌的DAPK甲基化率明显高于腺癌(分别为80.0%和16.7%),其差异有统计学意义(P<0.001).DAPK启动子甲基化和DAPK蛋白表达之间呈负相关(r=-0.849,P<0.001).结论:DAPK启动子CpG岛甲基化可导致基因失活,并可能参与宫颈癌的发生.     

PROMOTER HYPERMETHYLATION OF p16 GENE AND DAPK GENE IN SERA FROM HEPATOCELLULAR CARCINOMA （HCC） PATIENTS

Objective： To analyze the aberrant methylation of p16 gene and DAPK gene in sera from primary liver cancer patients ad to evaluate the clinical significance. Methods： A methylation-specific PCR was performed for the detection of promoter hypermethylation of p16 gene and DAPK gene in blood DNA from 64 cases of HCC patients, and to analyze the relation of the aberrant methylation of p16 gene and KAPK gene and the clinical pathological data. Results： 76.6%（49/64） of the sera from 64 cases of HCC patients showed hypermethylation for p16 promoter and 40.6% （26/64） for KAPK promoter, whereas no methylated p16 gene promoter and DAPK gene promoter were found in sera from benign liver diseases patients and normal control. Methylated p16 gene and KAPK gene promoters in sera did not strongly correlated with HBsAg, stage, metastasis and differentiation in HCC; but strongly correlated with AFP. Conclusion： Detection of the aberrant methylation of p16 gene and KAPK gene in blood DNA from HCC patients might offer an effective means for the earlier auxiliary diagnosis of the malignancy.     

中国非小细胞肺癌患者血清DAPK基因、p16基因启动子甲基化的分析

Objective： To evaluate the clinical significance of the aberrant methylation of DAPK gene and p16 gene in sera from 65 NSCLC patients from Nanjing General Hospital of Nanjing Command, China. Methods： A methylation-specific PCR （MSP） was performed for the detection of promoter hypermethylation of DAPK gene and p16 gene in blood DNA from 65 cases of NSCLC, and to analyze the relation of the aberrant methylation of DAPK gene and p16 gene and the clinicopathological data. Results： 30.8% （20/65） of the sera from 65 cases of NSCLC showed hypermethylation for DAPK promoter and 43.1% （28/65） the same for p16 promoter, whereas no methylated DAPK gene promoter and p16 gene promoter were found in sera from the patients with lung benign diseases and normal controls. Methylated DAPK gene promoter and p16 gene promoter in sera were not closely correlated with the pathological classification, stage, metastasis and differentiation in NSCLC. Conclusion： Detection of the aberrant methylation of DAPK gene and p16 gene in blood DNA from NSCLC patients might offer an effective means for the earlier auxiliary diagnosis of the malignancy.     

食管鳞癌中p16基因启动子区甲基化及其表达

 目的探讨食管鳞癌（ESCC）p16基因甲基化的状况及其表达与食管鳞癌临床病理特征之间的关系。方法采用甲基化特异性PCR方法（MSP）分别检测75例食管癌组织、癌旁组织和切缘组织p16基因启动子区域CpG岛甲基化状态。采用Envision免疫组化法检测食管癌组织及癌旁组织的p16蛋白的表达。结果75例标本中，食管癌组织、癌旁组织和切缘细织p16基因甲基化率分别为41．3％（31／75）、13．3％（10／75）和6．67％（5／75）。癌组织和癌旁组织P16蛋白的阳性表达率分别为29．3％（22／75）和56．7％0（17／30）。31例癌组织p16基因甲基化阳性标本中有2例（6．4％）检测到P16蛋白的表达，而44例癌组织p16基因甲基化阴性标本中有20例（45．5％）检测到P16蛋白的表达。食管癌组织p16基因甲基化率显著高于癌旁组织和切缘组织（P〈0.01），P16蛋白表达与p16基因甲基化呈负相关。p16基因启动子区甲基化与食管癌的组织学分级、肿瘤部位无明显相关，与临床分期、淋巴转移密切相关。结论p16基因甲基化在食管癌发生发展中起着重要作用，食管鳞癌的分期和淋巴结转移与p16基因甲基化之间有密切关系。     

胰腺组织p15、p16抑癌基因CpG岛甲基化研究

Objective： To detect the aberrant methylation patterns in the CpG islands of p16 and p15 tumor suppressor genes, and to analyze its correlation with pancreatic carcinogenesis and with clinicopathological characteristics of patients with pancreatic cancer （PC）. Methods： The methylation-specific polymerase chain reaction （MSP） method was used to monitor methylation patterns in the CpG islands of p15 and p16 genes from 29 cases of PC and 3 cases of chronic pancreatitis （CP） paraffin-embedded tissue, as well as 2 cases of normal liver tissues and 12 cases of normal blood samples. Results： p15 and p16 genes were detected to show unmethylation patterns and no amplification using methylation-specific primers in control group. The aberrant methylation rates of p16 in carcinoma tissue and adjacent noncarcinoma tissue were 37.9% （11 of 29 cases） and 34.5% （10 of 29 cases） respectively. Of the 11 aberrant methylated samples, 5 showed complete methylation and 6 hemimethylation. The methylation rates of p15 gene in carcinoma tissue and adjacent noncarcinoma tissue were 27.5% （8/29） and 24.4% （7/29） respectively. Of the 8 aberrant methylated samples, 3 showed complete methylation and 5 hemimethylation. In 6 PC samples, aberrant methylation in CpG islands of both p15 and p16 genes existed simultaneously. The aberrant methylation patterns in CpG islands of p15 and p16 genes had no close correlation with the clinicopathological characteristics （age, sex, smoking, volume of primary tumor, differentiation, clinical stage and histological classification） of the patients with PC （P〉0.05）. Conclusion： The aberrant methylation in CpG islands of p15 and p16 genes could be regarded as an early molecular event in PC and had no close correlation with the clinicopathological characteristics of the patients with PC.     

Concurrent hypermethylation of multiple regulatory genes in chewing tobacco associated oral squamous cell carcinomas and adjacent normal tissues

The methylation pattern in the promoter region of p16, DAPK, MGMT and GSTP1 genes was investigated in oral cancer tissues and tumor associated adjacent tissues, using methylation specific PCR assay. The samples constituted 60 primary oral tumors and corresponding adjacent clinically and histopathologically normal mucosa, and buccal epithelial scrapings from 20 normal healthy individuals without any tobacco habits. The incidence of hypermethylation in oral tumor and adjacent mucosa for p16 gene was 66.7 and 50%, for DAPK was 68.3 and 60%, and MGMT gene was 51.7 and 26.7%, respectively. The overall hypermethylation in the three genes in the primary tumor was 86.7%, and corresponding adjacent normal mucosa tissues 76.7%. Hypermethylation was not observed in the promoter region of GSTP1 gene in either the primary tumors or the corresponding adjacent normal mucosa. Absence of aberrant methylation in the four genes was noted in buccal scrapings from normal healthy individuals with no tobacco habits. Thus, a high frequency of promoter region hypermethylation was observed in p16, DAPK and MGMT genes in oral cancer tissues as well as in corresponding adjacent normal mucosa. Our results indicate that epigenetic alteration of these genes is a frequent event in oral cancer, and is an early event observed in normal oral mucosa of the patients, indicating the critical importance of the epigenetic alteration in chewing tobacco associated oral carcinogenesis.