肺癌患者癌组织和痰液细胞中p53和K-ras基因突变的研究

目的 检测肺癌组织和肺癌患者痰液脱落细胞中ｐ５３、Ｋ－ｒａｓ基因突变情况,比较联合检测ｐ５３、Ｋ－ｒａｓ和单一检测ｐ５３或Ｋ－ｒａｓ基因在肺癌诊断中的价值。方法 应用ＰＣＲ－ＳＳＣＰ－银染法检测了５９例肺癌组织、癌旁肺组织、１４全肺部良性病变肺组织及患者痰液脱落细胞中ｐ５３基因第５～８外显子、Ｋ－ｒａｓ基因第１外显子突变。结果 肺癌组织中ｐ５３基因突变率为３７．３％（２２／５９）,痰液脱落细胞为３     

人肺癌组织中C—Ha—ras,C—myc和C—sis基因的扩增

用同位素α－＾３２ＰｄＣＴＰ标记癌基因Ｃ－Ｈａ－ｒａｓ，Ｃ－ｍｙｃ和Ｖ－ｓｉｓ基因片段，通过点杂交和Ｓｏｕｔｈｅｒｎ杂交法检测１７例鳞癌，９例腺癌６例其它类肺癌组织和６例非肺癌组织中Ｃ－Ｈａ－ｒａｓ，Ｃ－ｍｙｃ和Ｃ－ｓｉｓ基因扩增的情况，结果发现：４６．８％（１５／３２）的肺癌组织有Ｃ－Ｈａ－ｒａｓ基因扩增，其中１０例（５８．８％）鳞癌，３例腺癌（３３．３％）出现Ｃ－Ｈａ－ｒａｓ基因扩增，２５％（     

肾癌组织中ras基因产物p21表达和K－ras基因点突变分析

目的研究ｒａｓ基因激活在肾癌发生中的意义。方法应用聚合酶链反应－单链构象多态性分析（ＰＣＲ－ＳＳＣＰ）和免疫组织化学检测肾癌组织Ｋ－ｒａｓ基因第１２位密码子突变及其基因产物ｐ２１蛋白的表达水平。结果４２例肾癌中６例Ｋ－ｒａｓ基因第１２位密码子点突变，１９例ｐ２１蛋白阳性表达，正常肾组织和癌旁组织均阴性。ｐ２１表达与肿瘤分期和分级有关。ｐ２１阳性患者的手术后生存率明显低于ｐ２１阴性患者。结论肾癌Ｋ－ｒａｓ基因第１２位密码子点突变并不常见，ｐ２１表达可能成为肾癌恶性程度和预后评价的指标。     

肺癌组织中p53,K—ras,p53,p16基因的改变

目的 探讨ｐ５３、Ｋ－ｒａｓ、ｐ１５、ｐ１６基因在肺癌组织中改变情况及多基因联合检测在肺癌诊断中的价值。方法 对５９例肺癌组织和１４例肺部良性组织,应用ＰＣＲ－ＳＳＣＰ－银染法检测ｐ５３基因第５～８外显子突变情况;应用ＰＣＲ－ＲＦＬＰ法对Ｋ－ｒａｓ基因突变进行检测;并应用ＰＣＲ法检测ｐ１５、ｐ１６基因纯合缺失情况。结果肺癌组织中ｐ５３、ｐ１５、ｐ１６基因改变频率分别为３７．３％、１１．９％、２３．     

非小细胞肺癌CT与相关基因表达的研究

目的：研究非小细胞（ＮＳＣＬＣ）肺癌ＣＴ征象与肺癌相关基因ｐ５３,Ｃ－ｍｙｃ,Ｋ－ｒａｓ,及Ｃ－ｅｒｂＢ２表达间的关系。方法：运用免疫组化法,检测５４例经手术病理证实的非小细胞肺癌组织中的ｐ５３,Ｃ－ｍｙｃ,Ｋ－ｒａｓ及Ｃ－ｅｒｂＢ２基因的表达,并分析其与术前ＣＴ征象间的关系。结果：ｐ５３表达与非小细胞肺癌期别及ＣＴ征象中瘤体大小,深分叶征,毛刺征,空洞征,及纵隔淋巴结转移有关,但与组织类型胸,胸     

非小细胞肺癌p53基因突变检测及临床病理分析

目的　探讨ｐ５３基因点突变在非小细胞肺癌中的发生情况及临床病理联系。方法　运用聚合酶链反应（ＰＣＲ）－单链构象多态性分析方法（ＳＳＣＰ）检测原发性非小细胞肺癌的ｐ５３ 基因第５－８外显子点突变。结果　８例良性肿瘤均无点突变发生,４０ 例肺癌中１９例（４７．５％ ）有点突变发生,点突变发生与临床分期和淋巴结累及程度明显相关（Ｐ＜ ０．０２）。结论　ｐ５３基因点突变在非小细胞肺癌的发生和进展中起着相当重要的作用。     

p53基因突变与人非小细胞肺癌临床病理生理分析

目的 探讨ｐ５３基因点突变与人非小细胞肺癌临床生理特征的联系。方法 应用聚合酶反应－单链构象多态性分析方法（ＰＣＲ－ＳＳＣＰ０检测原发性非小细胞肺癌癌组织的ｐ５３基因第５～８外显子点突变。结果 ４０例肺癌组织中１９例（４７．５％）有点突变发生,８例良性肿瘤组织均无ｐ５３基因点突变发生,点突变发生与病理分期和淋巴结转移有明显关系（Ｐ〈０．０２５）,结论ｐ基因估变在非小细胞肺癌的发生和进展中可能起重要作用。     

癌基因K-ras、C-myc、erbB-2的激活与非小细胞肺癌组织学类型相关性研究

为探讨癌基因突变、扩增与非小细胞肺癌发生及类型间的关系，应用核酸杂交的方法检测了63伤1肺手术标本中（腺癌19例，鳞癌23例，腺鳞癌3例，大细胞癌3例，小细胞癌3例，其它12例）三种癌基因（c－myc、K－ras、erbB－2）的扩增及突变。实验结果显示：50％（24／48）的非小细胞癌分别出现三种癌基因的激活；31．6％（6／19）的腺癌出现K－ras12密码子突变及K－ras扩增；30．4％（7／23）的鳞癌有erbB－2扩增；10．4％（5／48）的非小细胞癌存在c－myc扩增。提示K－ras突变可能为肺腺癌所特有，而erbB－2扩增是肺鳞癌发生的重要因素。     

人肿瘤发生中的多发性遗传改变

人肿瘤发生中的多发性遗传改变１．多发性遗传改变一种以上与癌相关基因的改变，也就是Ｎ－ｒａｓ的激活和ｃ－ｍｙｃ的放大，是由Ｗｅｉｎｂｅｒｇ和他的同事在ＨＬ－６０培养细胞中首先发现的．紧接着，Ｔａｙａ等阐述了在人肺巨细胞癌即Ｌｕ－６５中Ｋ－ｒａｓ突变和ｃ...     

原发性肝癌组织癌基因ras和抑癌基因P~（53）点突变研究

应用多聚酶链延伸反应──限制性片段长度多态性分析法（ＰＣＲ－ＲＦＬＰ）对４３例福尔马林液固定、石蜡包埋肝癌组织癌基因ｒａｓ和抑癌基因Ｐ（５３）点突变进行分析。结果表明，ｒａｓ基因总突变率为４１．９％（１９／４３），其中Ｎ－ｒａｓ第１２位密码子突变率为３７．２％（１６／４３），Ｃ－Ｋｉ－ｒａｓ第１２位密码子突变率为６．９％（３／４３），Ｃ－Ｋｉ－ｒａｓ第１３位、Ｃ－Ｈａ－ｒａｓ第１２和６１位密码子未发现有突变者。Ｐ~（５３）基因第２４９密码子突变率为２０．９％（９／４３），第２４８密码子未发现有突变者。结果提示Ｎ－ｒａｓ第１２密码子和Ｐ（５３）第２４９密码子突变与肝癌的发生和发展有关。     

大鼠肺鳞癌癌变过程中凋亡和增殖相关基因表达的研究

背景与目的:细胞凋亡信号转导的关键是凋亡下游蛋白caspase-3的激活。有关caspase-3与人类非小细胞肺癌的研究已取得了很大的进展,但它与大鼠肺鳞癌的关系的研究未见报道。本研究探讨大鼠肺鳞癌癌变过程中细胞增殖与凋亡相关基因cas-pase-3和增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的表达在癌变过程中的作用。方法:取60只Wistar大鼠,其中50只用化学致癌物3-甲基胆蒽(3-methylcholanthrene,MCA)及二乙基亚硝胺(diethyinitrosamine,DEN)碘油溶液于左肺叶支气管灌注以诱发大鼠肺鳞状细胞癌;另10只灌注碘油作为对照。用免疫组织化学SP法检测癌变过程中caspase-3和PCNA蛋白的表达;TUNEL法检测凋亡细胞。结果:对照组大鼠支气管粘膜上皮、癌前病变和肺鳞癌中,caspase-3蛋白表达的阳性评分均值分别为3.10±0.99、2.25±1.13、1.38±0.95;PCNA的平均增殖指数(PCNA-LI)分别为:14.10±5.02、28.13±8.72、41.88±14.24;平均凋亡指数(AI)分别为:0.60±0.52、2.06±0.85、2.26±1.14。肺鳞癌与对照组大鼠支气管粘膜上皮相比,其caspase-3蛋白阳性表达的差异有非常显著性(P<0.01),与癌前病变相比其caspase-3阳性表达的差异有显著性(P<0.05)。对照组大鼠支气管粘膜上皮为低增殖指数和低凋亡指数,分别与后     

K-ras activation in non-small cell lung cancer in the dog.

To investigate the role of K-ras mutations in canine non-small cell lung cancer, we first determined the nucleotide sequence of the normal canine K-ras gene and then examined 21 canine lung tumors for activating K-ras mutations. Canine K-ras was analyzed by direct sequencing of polymerase chain reaction products generated with oligonucleotide primers derived from the human K-ras sequence. Four nucleotide differences were found between the canine and human K-ras sequence from position 5 to 211. The deduced amino acid sequence of the canine gene was identical to that of the human. Activated K-ras alleles were detected in 5 of the 21 canine lung tumors examined. The activating lesions were point mutations, predominantly in codon 12. Of the 14 adenocarcinomas examined, 2 (14%) had K-ras mutations. Two of 5 (40%) adenosquamous carcinomas and the only large cell carcinoma also contained activated alleles. The overall frequency of K-ras point mutation in non-small cell lung cancer (25%) is similar to that reported in human non-small cell lung cancer. We conclude that K-ras activation by point mutation is associated with, but not necessary for, non-small cell lung cancer development in the dog.     

Postmenopausal estradiol-progestagen therapy and risk for uterine cervical cancer

The aim of this study was to evaluate the association of postmenopausal estradiol-progestagen therapy (EPT) with the risk for precancerous lesions, squamous cell carcinoma and adenocarcinoma of the uterine cervix. All Finnish women who had used EPT in 1994-2008 for at least 6 months (n = 243,857) at the age of 50 years or more were identified from the national Medical Reimbursement Registry and linked to the Finnish Cancer Registry. The incidence of cervical precancerous or cancerous lesions among EPT users was compared to that in the background population. There were 210 EPT users with squamous lesions (178 with precancerous and 32 with cancer) and 79 EPT users with glandular lesions (14 precancerous and 65 adenocarcinomas). The ever use of EPT did not associate with the incidence of precancerous lesions, but the risk for squamous cell carcinoma decreased (standardized incidence ratio 0.41; 95% confidence interval 0.28-0.58) and that for adenocarcinoma increased (1.31; 1.01-1.67). After the use of EPT for 5 years, the risk for squamous cell carcinoma decreased (0.34; 0.16-0.65), and the risk for adenocarcinomas increased (1.83; 1.24-2.59). The prolonged use of EPT is associated with the occurrence of cervical malignancies. If the association would be a causal one, the use for 5+ years among 10,000 women followed for 10 years would mean about two to three fewer cases of cervical squamous cell carcinoma but about two extra cases with adenocarcinoma.     

Increased epidermal growth factor receptor (EGFR) gene copy number is strongly associated with EGFR mutations and adenocarcinoma in non-small cell lung cancers: a chromogenic in situ hybridization study of 182 patients

SUMMARY: To evaluate the association of epidermal growth factor receptor (EGFR) gene copy number with EGFR and k-ras mutation status and tyrosine kinase inhibitor (TKI) sensitivity in non-small cell lung cancer (NSCLC), EGFR gene copy number of 182 NSCLC tumor specimens were analyzed by chromogenic in situ hybridization (CISH). EGFR and k-ras mutation analyses were also performed for, respectively, 176 and 157 of the 182 patients. Additionally, 36 patients in this study had received TKI monotherapy. The tumor was considered to be CISH positive if the gene copy number was >or=5 signals per nucleus in >or=40% of tumor cells. CISH-positive tumors were strongly associated with adenocarcinoma (56.8%) compared with squamous cell carcinoma (15.9%) (p<0.0001). The CISH-positive tumors were also strongly associated with EGFR mutations (78%) compared with wild type (20.2%) (p<0.0001). Only six tumors had k-ras mutations. None had EGFR mutation and only one was CISH positive. In the patients treated with TKI, EGFR mutation was strongly associated with TKI responsiveness (22/25 responders) (p<0.0001), but the CISH-positive tumors were only marginally significant (18/25 responders) (p=0.0665). Patients with EGFR mutations or CISH-positive tumors were all associated with longer median survival, although not statistically significant. Our results suggest Increased EGFR copy number was highly correlated with EGFR mutation in adenocarcinoma. Although it is less correlated with TKI responsiveness when compared with EGFR mutations, it still could be a good alternative molecular predictive marker for TKI responsiveness, since CISH can be done on paraffin section and is much quicker than DNA sequencing.     

Frequent polymorphic variations but rare tumour specific mutations of the S100A2 on 1q21 in non-small cell lung cancer

Contrary to the recent hypothesis that S100A2 is a tumour suppressor, no somatic mutations have yet been identified. We therefore screened 90 non-small cell lung carcinoma (NSCLC) samples, initially for mutations in S100A2 and then also for mutations in P53 and K-RAS genes. Alterations were detected in 46.7% of squamous lung cancer (SCC) samples, but we detected only one novel tumour specific mutation, Q23X in squamous carcinoma. We detected four polymorphisms, two of them published for the first time (144+109 C/G and 297+75A/G) and two already published: S62N, in the coding region and related to squamous cell carcinoma (SCC), and 297+17T/C. Analysis of S100A2 expression revealed that expression in adenocarcinomas and squamous cell carcinomas is significantly different, but not related to any of the found alterations. In one tumour with S62N polymorphism, P53 and K-RAS genes were also mutated, while two tumours with the Q23X mutation have a P53 but no K-RAS mutation. To the best of our knowledge, this is the first report describing alterations in the S100A2 gene proving a relation between changes in predominantly squamous lung cancer.     

CD44变异型在支气管粘膜上皮癌变过程中的表达及其意义

目的　探讨CD44变异型 (以下简称CD44v)表达在肺鳞癌发生中的作用。方法　应用免疫组织化学方法检测 3 4例肺鳞癌及其癌前病变标本中CD44v的表达 ,分析CD44v在肺鳞癌发生过程中出现表达的时间和频率。结果　癌旁正常支气管上皮细胞未见CD44v表达 ,在鳞状化生的支气管上皮细胞、轻→中→重度不典型增生、原位癌、浸润癌中 ,CD44v表达的阳性率逐渐升高。结论　本研究结果表明 ,CD44v可作为判断肺鳞癌发生可能性的标记物 ,在肺鳞癌的早期诊断中具有重要意义。     

K-ras oncogene mutation as a prognostic marker in non-small cell lung cancer: a combined analysis of 881 cases.

The treatment of non-small cell lung cancer (NSCLC) remains unsatisfactory, with most patients succumbing to metastatic disease within 5 years of diagnosis. Improved selection of patients for aggressive adjuvant therapy may contribute to improved survival. Mutation of the k-ras oncogene is considered a potentially clinically useful prognostic biomarker for this purpose. This report presents the results of a meta-analysis performed to determine whether the existing data support such a role for k-ras mutations in NSCLC. Two year survival data derived from 881 NSCLC patients in eight published studies were analyzed using a general variance-based meta-analytical method employing confidence intervals. The outcome of interest was a summary relative risk (RR(s)) reflecting the risk of death at 2 years associated with k-ras mutation-positive versus k-ras mutation-negative disease. Prior to calculation of RR(s), analysis for heterogeneity showed Q to equal 15.52 (7 df, P = 0.03). This indicated heterogeneity across the analyzed studies in terms of their estimate of effect. Possible sources of heterogeneity were identified and included selection bias and various other sources of uncontrolled confounding. Although a RR(s) of 2.35 (95% CI = 1.61-3. 22) was found when all eight studies were combined (favoring a negative prognostic role for k-ras mutation), it is unclear whether the magnitude of the RR(s) would persist after adjusting for other well-established prognostic indicators (e.g. stage). The existing data suggest that k-ras mutation may be associated with shortened survival in NSCLC, although this finding awaits confirmation in well-designed multivariate analyses which adjust for the effect of known prognostic indicators.     

非小细胞肺癌中3p^25杂合性丢失研究

目的为探讨非小细胞肺癌（ＮＳＣＬＣ）细胞中第三号染色体短臂上３ｐ２５位点上等位基因的杂合性丢失（ＬｏｓｓｏｆＨｅｔｅｒｏｚｙｇｏｓｉｔｙＬＯＨ）与肺癌发生发展的相关性。方法采用银染聚合酶链反应（ＰＣＲ）结合二核苷酸（ＣＡ）ｎ重复程序出现多态性评价杂合性丢失方法，分析５８例ＮＳＣＬＣ组织中３ｐ２５等位基因的杂合性丢失。结果５８例肺癌组织中有２９例出现３ｐ２５ＬＯＨ，总的杂合性丢失率为５０％。３ｐ２５的杂合性丢失率在１８例腺癌中为１２例（６６．７％），明显高于鳞癌（４２．９％，１５／３５），有显著性统计学意义（Ｐ＜０．０５），但３ｐ２５杂合性丢失与ＮＳＣＬＣ临床分期关系不明显，Ⅰ期为４２．１％，Ⅱ期为５５．６％和Ⅲ期为５２．２％，５例正常胎儿肺组织均不出现３ｐ２５ＬＯＨ。结论３ｐ２５杂合性丢失在ＮＳＣＬＣ中普遍存在，从而说明３ｐ２５位点处存在着某些与肺癌发生发展有关的抑癌基因。