RASSF1A基因甲基化与非小细胞肺癌预后的相关性研究

背景与目的研究发现在很多肿瘤中均存在RASSF1A基因启动子区域高甲基化状态导致基因表达失活的现象,本研究就RASSF1A基因启动子的甲基化状态与非小细胞肺癌预后的关系进行探讨。方法采用甲基化特异的PCR检测150例非小细胞肺癌和20例肺部良性病变RASSF1A启动子甲基化状态。结果150例非小细胞肺癌中58例发现RASSF1A启动子存在甲基化(58/158,38.7%),20例肺部良性病变中无一例发现RASSF1A启动子甲基化。存在RASSF1A启动子高甲基化的病例预后较未发现RASSF1A甲基化的病例差(P=0.004),Cox回归分析显示RASSF1A启动子的甲基化状态是非小细胞肺癌术后的一个预后相关因素(RR=1.584,95%CI:1.040-2.411,P=0.032)。结论MSP法检测RASSF1A启动子甲基化状态可以作为非小细胞肺癌术后的一个预后评价指标。     

宫颈癌组织中RASSF1A基因启动子甲基化对其表达水平的影响及临床意义

目的 研究宫颈癌组织中RAS相关区域家族1A基因(RASSF1A)启动子甲基化水平和RASSF1A基因mRNA表达水平,分析其与宫颈癌临床病理参数的关系及临床意义.方法 收集40例宫颈癌组织及相应癌旁组织,采用巢式特异性甲基化方法检测RASSF1A基因启动子甲基化水平,采用实时荧光定量PCR(qRT-PCR)检测宫颈癌和癌旁组织中RASSF1A基因mRNA表达水平.结果 宫颈癌组织中RASSF1A基因mRNA表达水平(0.26±0.05)显著低于癌旁组织(0.28±0.03),差异有统计学意义(t=2.27,P=0.026);宫颈癌组织中RASSF1A基因启动子区的甲基化率(0.71%±0.04%)显著高于癌旁组织(0.66%±0.03%),差异有统计学意义(t=6.78,P=0.000);RASSF1A基因mRNA表达水平与病理分化程度(t=3.31,P=0.002)、国际妇产科联合会(FIGO)分期(t=2.13,P=0.040)、淋巴结转移(t=2.56,P=0.015)、浸润深度(t=2.93,P=0.006)有关;RASSF1A基因启动子区的甲基化水平与病理分化程度(t=2.08,P=0.045)、FIGO分期(t=2.66,P=0.011)、淋巴转移(t=2.22,P=0.033)、浸润深度(t=2.12,P=0.041)有关.结论 宫颈癌组织中RASSF1A基因启动子甲基化和RASSF1A基因mRNA的表达水平与宫颈癌的恶性程度相关,RASSF1A基因甲基化水平有望成为宫颈癌转移风险的重要指标.     

宫颈鳞癌组织中RASSF1A基因甲基化状态及其临床意义

背景与目的:Ras相关区域家族1A(ras associated domain family 1A,RASSF1A)基因是一种肿瘤候选抑痛基因,其启动子区高甲基化与多种人类上皮性肿瘤的发生有关.本研究通过检测宫颈鳞癌(cervical squamous cell carcinoma,CSCC)组织中RASSF1A基因启动子区甲基化状态及其蛋白表达情况,旨在分析RASSF1A基因甲基化与基因失活的相关性,并探讨RASSF1A基因甲基化与CSCC发生的关系.方法:分别采用甲基化特异性PCR(methylation-specific PCR,MSP)及免疫组织化学方法检测15例正常宫颈组织和46例CSCC组织中RASSF1A基因的甲基化程度及RASSF1A蛋白的表达情况,并分析RASSF1A基因甲基化与RASSF1A蛋白表达及其与CSCC临床病理特征之间的关系.结果:正常宫颈组织中未见RASSF1A基因甲基化,而CSCC组织中RASSF1A基因甲基化率为43.5%(20/46),两者比较差异具有统计学意义(P＜0.05).CSCC中临床Ⅰ期、Ⅱ期、Ⅲ～Ⅳ期的甲基化阳性率呈逐渐增高趋势(19.0%→58.8%→75.0%,P＜0.05),不同组织学分级、有无淋巴结转移与RASSF1A基因甲基化间未见明显相关性(P＞0.05).RASSF1A基因甲基化与RASSF1A蛋白的表达呈负相关(r=-0.469,P＜0.05).结论:RASSF1A基因启动子区甲基化是导致RASSF1A蛋白低表达的机制之一,可能与CSCC的发生发展有关.     

非小细胞肺癌患者血清Ras相关区域家族1A基因启动子异常甲基化检测及其临床意义

目的 研究非小细胞肺癌(NSCLC)患者血清Ras相关区域家族1A(RASSF1A)基因启动子区域的甲基化状态及其临床意义.方法 采用甲基化特异性聚合酶链反应(MSP)技术,检测75例NSCLC患者血清RASSF1A基因启动子区域的甲基化状态,并分析其与临床病理参数之间的相关性.结果75例NSCLC患者血清RASSF1A基因启动子区域异常甲基化检出率为30.7%(23/75),而35例肺部良性疾病患者和15例健康志愿者中检出率均为0,差异有统计学意义(P＜0.001).RASSF1A启动子异常甲基化与NSCLC患者的年龄、性别、病理类型无显著相关性,但在晚期及肿瘤分化程度较低的患者中检出率较高(P＜0.05).结论 RASSF1A启动子异常甲基化在NSCLC的发生、发展中起重要作用,有望成为NSCLC辅助诊断和预后判断的分子标记.     

食管癌组织与外周血RASSF 1A甲基化的检测及其临床意义

目的:通过对食管癌组织及同一患者对应的术前外周血中RASSF1A基因甲基化的检测,加深食管癌变分子机制的了解并为食管癌早期诊断和高危人群预警提供候选指标.方法:采用甲基化特异性PCR方法,分别检测来自食管癌高发区的30例食管癌患者血浆、肿瘤组织及癌旁正常组织中RASSF1A基因启动子甲基化状况.结果:食管癌组织RASSF1A甲基化阳性率为40%(12/30),而这12例癌组织甲基化阳性的患者其外周血甲基化阳性共7例,癌组织和外周血RASSF1A甲基化阳性一致率为58%(7/12),18例癌组织甲基化阴性的患者其外周血也均为阴性,阴性一致率为100%(18/18).癌旁正常食管组织甲基化率为13%(4/30),明显低于癌组织(40%,12/30),P<0.05.7例外周血甲基化阳性的患者中,淋巴结转移阳性5例(55%,5/9),阴性2例(10%,2/21),两者差异有统计学意义,P<0.05.低分化鳞癌的RASSF1A甲基化阳性率(83%,5/6)明显高于中分化鳞癌(24%,5/21),P<0.05.结论:外周血RASSF1A甲基化可以反映同一个体食管鳞癌组织中RASSF1A基因甲基化的状态,可能是高危人群筛查重要候选分子标志之一.     

宫颈癌细胞系RASSFIA基因启动子及第1外显子区甲基化状态的研究

背景与目的：抑癌基因Ras相关区域家族1A (Ras association domain family 1A,RASSF1A)启动子及第1外显子区CG位点高甲基化导致该基因沉默与多种恶性肿瘤的发生发展相关。本研究旨在探讨宫颈癌细胞系RASSFIA基因启动子及第1外显子区甲基化状态以及甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(5-Aza-2’deoxycytidine,5-Aza-dc)作用对RASSFIA基因表达的影响。方法：采用5 μmol/L(低浓度)和10 μmol/L(高浓度)的5-Aza-dc作用于HeLa、Caski、HT-3以及C-33A等4种宫颈癌细胞系,分别采用甲基化特异PCR (methylation-specific PCR,MSP)和亚硫酸盐基因组测序法(bisulfite genome sequencing,BGS)检测5-Aza-dc处理前后RASSF1A基因启动子及第1外显子区甲基化状态,RT-PCR检测干预前后RASSF1A基因mRNA的转录表达。结果：HeLa和Caski两种HPV阳性细胞系RASSF1A基因启动子及第1外显子区均呈低甲基化状态,mRNA表达阳性。低浓度和高浓度5-Aza-dc作用后,mRNA表达未见明显改变(F_HeLa=3.003,P=0.125;F_Caski=0.045,P=0.956)。HT-3和C-33A两种HPV阴性宫颈癌细胞系RASSF1A基因启动子及第1外显子区则表现为高度甲基化状态,mRNA表达受到抑制。低浓度和高浓度5-Aza-dc作用后,HT-3和C-33A细胞系RASSF1A基因启动子及第1外显子区CG位点甲基化程度降低,检测到其mRNA表达,高浓度5-Aza-dc作用组表达水平明显高于低浓度组和细胞对照组(F_HT-3=18.002,P=0.03;F_C-33A=17.179,P=0.03),LSD-t检验显示差异有统计学意义(P<0.05)。结论：HPV阳性和HPV阴性宫颈癌细胞系中RASSFIA基因启动子及第1外显子区甲基化状态不同;RASSF1A基因启动子及第1外显子区的高甲基化可抑制该基因表达;5-Aza-dc处理可使RASSF1A基因启动子及第1外显子区去甲基化,重新激活基因的表达,这种作用在一定范围内有剂量依赖性。     

急性髓系白血病RASSF1A基因启动子区异常甲基化临床意义研究

目的 RASSF1A基因异常甲基化可能参与血液肿瘤的发生,并为微小残留疾病(minimal residual de-sease,MRD)监测、分层、预后评估及靶向治疗提供依据.本研究旨在分析RASSF1A基因启动子区异常甲基化在急性髓系白血病(acutemyeloid leukemia,AML)中的临床意义.方法 选取2005-01-01-2013-03-01解放军总医院(113例)以及第一附属医院(39例)住院患者和门诊体检患者,共152例AML患者骨髓标本以及15例健康供者骨髓标本纳入本项研究.提取基因组DNA,并进行DNA硫化修饰;设计重亚硫酸盐测序PCR(bisulfite sequencing PCR,BS-PCR)引物以及甲基化特异性PCR(methylation specific PCR,MS-PCR)引物,进行PCR扩增,进而电泳分析以及DNA序列分析.同时对RASSF1A高甲基化组以及低甲基化组的血液学特点、骨髓原始细胞比例、细胞遗传学异常、基因异常、完全缓解率和总生存期进行统计学分析.结果 MS-PCR分析结果显示,RASSF1A基因在15例健康人中呈完全非甲基化状态,在152例AML患者中有38例出现启动子区高甲基化状态,其甲基化阳性率为25％.4例MS-PCR阳性AML患者经BS-PCR测序分析后,显示RASSF1A甲基化率分别为88.2％、85.5％、78.6％和92.7％,而在4例MS-PCR阴性患者RASSF1A基因启动子区甲基化率分别为10％、11.8％、12.7％和6.8％,4例健康供者RASSF1A基因启动子区甲基化率分别为5.0％、9.1％、8.2％和7.3％.进而通过统计学分析发现携带RASSF1A基因高甲基化的AML患者易合并存在ASXL1基因突变或DNMT3A基因突变.携带RASSF1A基因高甲基化的AML患者,其无进展生存期以及总生存期较短.结论 RASSF1A基因启动子区异常甲基化可能参与AML的发生,同时可能为AML分层诊治以及预后评估提供分子理论依据.     

血浆Ras相关区域家族蛋白1A基因甲基化在肝细胞癌分子诊断中的价值

目的:建立一种可检测微量DNA标本中DNA甲基化的甲基化敏感性限制性内切酶-定量PCR(methylation-sensitive restriction enzymes-based quantitative PCR,MSRE-qPCR)方法,并运用该技术探讨血浆Ras相关区域家族蛋白1A(Ras association domain family1A,RASSF1A)基因甲基化检测在肝细胞癌(hepatocellutar carcinoma,HCC)非侵入性诊断中的价值。方法:用MSRE HhaⅠ消化DNA样品,再用qPCR技术分析酶切结果,建立检测RASSF1A基因甲基化的MSRE-qPCR方法。以45例肝组织(20对HCC患者手术切除肿瘤标本及匹配非癌组织和5例正常肝)为材料,测试该方法的应用价值;运用亚硫酸氢盐测序PCR(bisulf ite sequencing PCR,BSP)技术进行进一步验证,并与甲基化特异性PCR(methylation specif ic PCR,MSP)方法相比较。再运用该技术检测150例血浆标本(包括72例HCC患者、37例肝硬化或慢性肝炎等良性病变患者和41例健康对照)的RASSF1A基因甲基化状态,并分析其与HCC患者临床病理参数的关系。结果:MSRE-qPCR法可定量检测低至1%以下的RASSF1A甲基化片段。20例HCC组织中有14例(70%)发生RASSF1A高甲基化,对应非癌组织中RASSF1A甲基化阳性率为25%,而5例正常肝组织均为阴性。MSRE-qPCR结果经BSP验证无误,且与MSP检测结果具有较好的一致性。HCC患者血浆RASSF1A甲基化阳性率(47/72,65.3%)显著高于健康对照(1/41,2.4%)和肝良性病变组(3/37,8.1%),差异均有统计学意义(P<0.0001)。联合检测血浆RASSF1A甲基化与血清AFP可显著提高HCC诊断效率。结论:建立的MSRE-qPCR方法要求样本少、操作简便、成本低廉,可定量检测RASSF1A基因甲基化水平。血浆RASSF1A甲基化分析对于HCC的非侵入性诊断具有重要价值。     

RASSF1A基因启动子外周血和组织甲基化在乳腺癌中的意义

目的通过对乳腺癌、癌旁组织及同一患者对应的术前外周血中RASSF1A基因甲基化的检测,加深对乳腺癌发病分子机制的了解并为乳腺癌早期诊断和预后判断提供候选指标。方法采用甲基化特异性PCR方法,分别检测156例乳腺癌患者血浆、肿瘤组织及癌旁正常组织和39例乳腺良性病变血浆及其正常组织中RASSF1A基因启动子甲基化状况。结果早期乳腺癌组织RASSF1A基因启动子甲基化发生率为62.2%(23/37),同一患者外周血甲基化发生率为56.7%(21/37),Kappa值为0.6553(P<0.001),40例血浆RASSF1A甲基化的患者中,37例发生淋巴结转移92.5%(37/40),3例未发生淋巴结转移7.5%(3/40),差异有显著性(P=0.0003);乳腺癌组织和外周血中的RASSF1A甲基化水平与乳腺癌的年龄、家族史、分型、分期、ER、PR、CerbB-2无关;晚期癌复发的86例中,其中癌组织甲基化病例的复发率为25.5%(13/51),癌旁组织甲基化的复发率为75.7%(53/70),两组差异有显著性(P<0.0001)。结论乳腺癌血浆RASSF1A甲基化与组织中的变化较为一致,可作为早期病例筛查和判断淋巴结转移的候选指标之一;晚期乳腺癌癌旁RASSF1A甲基化可能参与肿瘤复发,可作为预测晚期病例预后的候选指标之一。     

乳腺癌APC基因甲基化状况及其蛋白表达

背景与目的:结肠腺瘤性息肉病基因(adenomatous polyposis coli,APC)蛋白表达产物是Wnt信号转导途径重要组成部分,该基因失活使β-catenin蛋白降解障碍,从而使Tcf/Lef激活,引起基因异常转录,最终产生癌变.启动子区甲基化是导致抑癌基因转录失活的重要机制.本研究探讨乳腺癌APC基因启动子1A区甲基化状态与其蛋白表达的关系,并分析APC基因异常甲基化与乳腺癌临床病理特征的关系.方法:应用甲基化特异性PCR和免疫组织化学方法检测76例乳腺癌及相应癌旁乳腺组织中APC基因启动子1A区甲基化状态以及蛋白表达.结果:癌旁乳腺组织中均未发现APC基因启动子1A区甲基化,乳腺癌组织中APC基因启动子1A区甲基化率为36.8%.癌旁乳腺组织中APC蛋白阳性率为100%,乳腺癌中APC蛋白阳性率为52.4%.乳腺癌组织中APG基因甲基化与APC蛋白表达呈负相关(r=-0.351,P＜0.05),与TNM分期呈正相关(r=0.335,P＜0.05).结论:乳腺癌发展过程中APC基因启动子1A区出现异常甲基化,是导致该蛋白表达缺失的主要原因,是导致该基因失活的重要机制之一.     

E-钙粘蛋白及PTEN基因编码蛋白与胃癌浸润转移

目的:观察抑癌基因PTEN蛋白和ECD在胃癌组织中的表达,探讨其与胃癌生物学行为及预后的关系。方法:以兔抗人PTEN多克隆抗体、鼠抗人ECD单克隆抗体,采用SABC免疫组化法,检测100例胃癌手术切除标本中拟测指标的表达。以χ2和Logrank检验对结果做统计学分析。结果:ECD、PTEN蛋白在非癌胃粘膜中均见表达;在胃癌组织中表达下调或缺失。ECD异常表达率为42.0%;弥漫型胃癌异常表达率(48.57%),明显高于肠型胃癌(26.67%),(P<0.05);ECD异常表达与浸润深度有关(P<0.05)。胃癌组织中PTEN蛋白缺失率为59%;弥漫型胃癌缺失率(65.71%)明显高于肠型胃癌(43.33%),(P<0.05);伴淋巴结转移的胃癌缺失率(64.47%)明显高于无淋巴结转移者(41.67%),(P<0.05);PTEN蛋白缺失的患者比阳性表达者预后差(P=0.0066)。65.85%PTEN阳性表达者同时伴ECD正常表达。结论:两种标志物与胃癌浸润转移有关,PTEN表达与胃癌患者预后密切相关。将两种指标联合检测,可作为正确判断胃癌患者预后,指导临床治疗的分子生物学指标。     

Cytologic classification of precancer and cancer of the endometrium and esophagus and of malignant lymphomas

The paper discusses cytological classifications of precancer and cancer of the endometrium, esophagus and malignant lymphomas presented by cytologists from five Soviet research institutes of oncology. The classifications were based on the data of 4400 cases in conformity with WHO histologic classifications.     

世界卫生组织骨质疏松症防治工作报告和防治建议

引 言 作为对第51号综合处理非传染性疾病预防与控制的世界卫生组织决议的反应,1998年7月WHO成立了致力于不断完善对骨质疏松预防和治疗策略的工作小组。小组成员来自世界各国致力于骨质疏松研究的知名专家。Harry K.Genant为本届主席。这一项世界范围内的骨质疏松教育计划旨在通过世界范围的研究,不断改善对骨质疏松的诊断水平和发展并完善对骨质疏松病人的合理治疗。其重点将以发展中国家为主。并为各国政府及其卫生部门和病人群体提供世界性有关骨质疏松症的总体的、完整的指导性资料。该项研究、教育计划的实施将由世界各国的骨质疏松症研究和治疗机构共同完成,并经权威学术机构、政府和非政府组织进行有针对性的回顾研究,最终由WHO审议通过。     

Schwannomas of Head and Neck and Review of Literature

Benign nerve cell tumours have been given various names like schwannoma, neurilemmoma, neurinoma, neurofibroma, spindle cell tumours etc. Extra cranial head and neck schwannomas usually present as solitary and well-demarcated lesions. The lesion can cause secondary symptoms, such as nasal obstruction, dysphasia, and hoarseness, depending upon the location of the lesion. Fine needle aspiration cytology, CT scans, and MRI may be of limited help in the diagnosis of schwannomas. The treatment is complete surgical excision of the benign tumour and postoperative histopathological examination establishes the final diagnosis.     

Quality of life and care in leukemia,myeloma and non-malignant disease. Opinions of patients and relatives,and effects of geography and time

In a questionnaire study 140 subjects answered 4200 questions in 1980 and 1986. They consisted of patients with myeloma, acute leukemia, lung carcinoma, and non-malignant disease and their relatives. In 22 additional cases the questionnaire was not answered. The results show that myeloma patients are less content with the general care than leukemia patients (P < 0.05). Similarly, relatives of deceased myeloma patients are less satisfied with the information given to them than relatives of deceased leukemia patients (P < 0.001). The information has improved with time, however, since the patients were more satisfied in 1986 than in 1980 (P < 0.001) and relatives of myeloma patients still alive were more satisfied than relatives of patients who had died earlier (P < 0.001).     

A comparative study of knowledge and awareness of colorectal and breast cancerA comparative study of knowledge and awareness of colorectal and breast cancer

Aims: To assess and compare knowledge and awareness of colorectal cancer and breast cancer in a sample of the general population. Methods: Eleven hundred visitors to six different outpatient clinics, in a University Hospital, were given a study-specific questionnaire, based on educational material from the British Association of Cancer United Patients (CancerBACUP). The questionnaire consisted of 12 statements on the incidence, presentation, detection, treatment and prognosis of colorectal and breast cancer. Results: One thousand and sixty-eight individuals returned the questionnaire. One thousand and four completed questionnaires were analysed. The mean age (SD) of respondents was 50.1 (17.2) years, and the male to female ratio was 2:3. Respondents had read more about breast than about colorectal cancer (60.3%vs 32.4%,P <0.0001, McNemar's test). The proportion of correct answers for each statement on breast cancer was higher than for answers to corresponding items on colorectal cancer. Mean overall scores (95% CI) for breast and colorectal cancer were 88.1 (86.9, 89.2) and 64.4 (62.5, 66.3) respectively, the mean difference (95% CI) being 23.7 (22.0, 25.5). Scores were higher for breast cancer irrespective of age or gender. Conclusion: There is a low level of understanding of colorectal cancer in the general population when compared to breast cancer. This highlights the importance of public education in this common cancer.     

Prospective study of fruit and vegetable consumption and incidence of colon and rectal cancers

BACKGROUND: Frequent consumption of fruit and vegetables has been associated with a reduced risk of colorectal cancer in many observational studies. METHODS: We prospectively investigated the association between fruit and vegetable consumption and the incidence of colon and rectal cancers in two large cohorts: the Nurses' Health Study (88 764 women) and the Health Professionals' Follow-up Study (47 325 men). Diet was assessed and cumulatively updated in 1980, 1984, 1986, and 1990 among women and in 1986 and 1990 among men. The incidence of cancer of the colon and rectum was ascertained up to June or January of 1996, respectively. Relative risk (RR) estimates were calculated with the use of pooled logistic regression models accounting for various potential confounders. All statistical tests were two-sided. RESULTS: With a follow-up including 1 743 645 person-years and 937 cases of colon cancer, we found little association of colon cancer incidence with fruit and vegetable consumption. For women and men combined, a difference in fruit and vegetable consumption of one additional serving per day was associated with a covariate-adjusted RR of 1.02 (95% confidence interval [CI] = 0.98-1.05). A difference in vegetable consumption of one additional serving per day was associated with an RR of 1.03 (95% CI = 0.97-1.09). Similar results were obtained for women and men considered separately. A difference in fruit consumption of one additional serving per day was associated with a covariate-adjusted RR for colon cancer of 0.96 (95% CI = 0.89-1.03) among women and 1. 08 (95% CI = 1.00-1.16) among men. For rectal cancer (total, 244 cases), a difference in fruit and vegetable consumption of one additional serving per day was associated with an RR of 1.02 (95% CI = 0.95-1.09) in men and women combined. None of these associations was modified by vitamin supplement use or smoking habits. CONCLUSIONS: Although fruits and vegetables may confer protection against some chronic diseases, their frequent consumption does not appear to confer protection from colon or rectal cancer.