158例非小细胞肺癌中3p25杂合性丢失研究

目的 探讨非小细胞肺癌组织（ＮＳＣＬＣ）中３ｐ２５位点上等位基因的杂合性丢失（ＬＯＨ）与肺癌发生，发展的相关性。方法 采用银染聚合酶链反应结合二核苷酸（ＣＡ）ｎ重复程序出现多态性评价ＬＯＨ，分析１５８例ＮＳＣＬ癌组织中３ｐ２５等位基因的ＬＯＨ。结果 １５８例ＮＳＣＬＣ组织中８０例出现３ｐ２５ＬＯＨ，总的杂合性丢失率为５０．６％，腺癌组织中３ｐ２５杂合性丢失率为６２．０％，鳞癌为４３．２％，两者间差     

非小细胞肺癌中3p14的杂合性丢失的研究

目的探讨非小细胞肺癌（ＮＳＣＬＣ）细胞中３ｐ１４位点等位基因的杂合性丢失（ｌｏｓｓｏｆｈｅｔｅｒｏｚｙｇｏｓｉｔｙ,ＬＯＨ）与ＮＳＣＬＣ发生及发展之间的相关性。方法采用ＰＣＲ结合二核苷酸重复序列多态性方法并经ＬＯＨ┐银染法检测７４例非小细胞肺癌新鲜手术切除标本。结果７４例ＮＳＣＬＣ组织中３１例出现３ｐ１４ＬＯＨ,总丢失率为４３．２％;其中鳞癌的杂合性丢失为１４例（３８．９％,１４／３６）,腺癌为１３例（４４．８％,１３／２９）,３ｐ１４杂合性丢失在Ⅰ,Ⅱ,Ⅲ期ＮＳＣＬＣ中分别为３１．６％（６／１９）,４２．９％（９／２１）,４７．４％（１６／３４）,而正常肺组织未见３ｐ１４ＬＯＨ。结论３ｐ１４杂合性丢失普遍出现于ＮＳＣＬＣ中,提示该位点可能存在着一个或多个抑癌基因,因此３ｐ１４丢失可能与肺癌的发生及发展相关     

肺癌染色体8p21～23区域精细缺失图谱的构建

目的：进一步明确肺癌在８ｐ２１￣２３区域等位基因杂合性丢失（ｌｏｓｓ ｏｆ ｈｅｔｅｒｏｚｙｇｏｓｉｇｔｙ,ＬＯＨ）的频率与共同缺失区范围,以便分离该区域内与肺癌相关的候选抑瘤基因。方法：应用位于８ｐ２１￣２３的１６个微卫星多态标记,对３２例肺癌组织和２例肺癌细胞系进行ＬＯＨ分析。结果：３２例肺癌组织与２例肺癌细胞系中共２０例（５８．８２％）存在至少一个位点的杂合性丢失,缺失频率最高的位点中８ｐ２     

原发性NSCLC中3p杂合性丢失的研究

目的 探讨非小细胞癌（ＮＳＣＬＣ）中第３屿染体短臂（３Ｐ）上两个独特区域－－３Ｐ１４和３Ｐ２５等位基因的杂合性丢失（ＬＯＨ）与ＮＳＣＬＣ发生、发展之间的关系。方法 采用聚合酶链反应（ＰＣＲ）结合二核苷酸（ＣＡ）ｎ重复序列多态性方法,分析１６２例鹇手术标本、８５例癌旁肺组织及５例肺部良性疾病组织中３Ｐ１４和３Ｐ２５位点的ＬＯＨ。结果 １６２例ＮＳＣＬＣ组织中共有１０８例（６６．６７％）出现ＬＯＨ,８     

鼻咽癌染色体3p21-26的等位基因杂合性丢失研究

目的细胞遗传学研究表明,３号染色体缺失是鼻咽癌常见的染色体异常之一。分子生物学研究表明,染色体３ｐ在鼻咽癌中出现高频率的等位基因杂合性丢失（ＬＯＨ）。本研究将进一步确定鼻咽癌３ｐ等位基因杂合性丢失的频率及范围。方法应用位于３ｐ２１２６区域１６个微卫星多态性位点,对２４例低分化鼻咽癌患者进行了ＬＯＨ分析。结果２４例患者中有１６例存在杂合性丢失（６６．７％）。丢失频率最高的两个位点是Ｄ３Ｓ１５６０（５０％,１１／２１）和Ｄ３Ｓ１６２０（５０％,９／１８）。在具有丢失的１６例患者中,８例显示为１个连续的多个相邻位点的杂合性丢失区域,５例患者存在２个或２个以上的杂合性丢失区。病例１,３,４,７,８,１０,１６,１７,１８,１９和２２在Ｄ３Ｓ１５９７和Ｄ３Ｓ１２９７之间,表现为一个不同大小的杂合性丢失区。结论最小共同丢失区位于Ｄ３Ｓ１５６０Ｄ３Ｓ１６２０（３ｐ２５．３２６．２）之间,提示该区域有一个尚未克隆的、与晚期鼻咽癌明显相关的抑癌基因。     

原发性肝细胞癌多染色体杂合性丢失研究

研究原发性肝细胞癌（ＨＣＣ）５对染色体３８个位点等位基因杂合性丢失（ＬＯＨ），ＬＯＨ与肝癌的临床病理改变和肝炎病毒的关系。方法：用ＰＣＲ微卫星多态性分析检测肝癌ＬＯＨ。结果发生高频率ＬＯＨ的位点有染色体１ｐ的Ｄ１Ｓ１８６（这，４８．１％）和Ｄ１Ｓ２４３（１ｐ３６．３，５１．６％）；染色体９ｎ２４的Ｄ９Ｓ５４（６１．８％），９这的Ｄ９Ｓ１７４７（５２．％）和ＤＤ９Ｓ１７５２（５１．８％）ＷＵＧ ＨＫ     

发性肝癌9号染色体等位基因杂合性丢失研究

目的：对广东地区３７例肝癌染色体９ｐ的９个位点进行微卫星多态性分析,明确这睦位点染色体等位基因杂合性丢失（ＬＯＨ）的情况,定位该区域可能存在的与肝癌有关的肿瘤抑制基因。方法：对所取位点用ＰＣＲ法进行微卫星多态性分析并确定发生ＬＯＨ的位点;结果：３０例（８０．８０％）在至少一个位点发生ＬＯＨ,其中ＬＯＨ频率最高的位点是Ｄ９Ｓ５４（６０．６％）,ＬＯＨ频率高于５０％的位点有位于９ｐ２１带的ＩＦＮＡ,Ｄ     

原发性非小细胞肺癌（NSCLC）中MTS2／p15基因丢失的研究

目的探讨ＭＴＳ２／ｐ１５基因缺失与人体非小细胞肺癌（ＮＳＣＬＣ）的发生及发展的相关性。方法采用聚合酶链式反应（ＰＣＲ）技术，分别检测１６０例不同病理类型及分期的肿瘤组织，１０例非癌性肺组织及２例胎肺组织中ｐ１５基因的纯合性丢失。结果１６０例ＮＳＣＬＣ组织中ｐ１５基因的丢失率以腺癌为最高６４．７％，鳞癌其次４３．９％，混合型鳞腺癌为最低３７．０％，１０例非癌性肺组织及２例胎肺组织均未见有ｐ１５基因的丢失。经ＴＮＭ分期分析表明，Ⅰ期ｐ１５基因丢失率为３４．６％，Ⅱ期为４７．４％，Ⅲ期为６１．４％，Ⅲ期的ｐ１５丢失率明显高于Ⅰ期（Ｐ＜０．００５）。结论ＭＴＳ２／ｐ１５基因缺失除与ＮＳＣＬＣ的发生相关外，还可能涉及到患者病程的进展。     

ras和erbB2基因在肺癌中的表达及意义

目的：了解ｒａｓ和ｅｒｂＢ２基因在肺癌中表达的临床意义。方法：采用免疫组化技术研究了ｒａｓｐ２１和ｅｒｂＢ２ｐ１８５在４０例肺癌（腺癌１９例，鳞癌１８例，腺鳞癌１例，小细胞肺癌２例）中的表达，并将肺癌按临床病理特征分组进行对比分析。结果：①在非小细胞肺癌（ＮＳＣＬＣ）中，ｐ２１的阳性率为６１％，ｐ１８５的阳性率为４２％，２例小细胞肺癌（ＳＣＬＣ）均无ｐ２１和ｐ１８５的表达。②ｐ１８５在肺腺癌中的表达明显高于鳞癌（Ｐ＜０．０５）。③在ＮＳＣＬＣ中，ｐ２１在Ⅰ～Ⅱ期中的阳性率为２５％，Ⅲａ～Ⅲｂ期中的阳性率为７７％，两者有非常显著的差异（Ｐ＜０．０１）。④ｐ２１和ｐ１８５阳性的ＳＮＣＬＣ患者，其淋巴结转移发生早，速度快（Ｐ＜０．０５）。⑤ｐ１８５在ＳＮＣＬＣ中的表达存在性别差异，女性明显高于男性（Ｐ＜０．０１）。结论：ｒａｓ和ｅｒｂＢ在ＮＳＣＬＣ中的表达具有重要的临床意义。     

卵巢癌及宫颈癌中17p13.3的杂合性丢失

目的探讨染色体１７ｐ１３．３的杂合性丢失（ＬＯＨ）与卵巢癌、宫颈癌发生及发展之间的相关性。方法采用ＰＹＮＺ．２２探针做Ｓｏｕｔｈｅｒｎ印迹技术,检测２４例卵巢癌、９例宫颈癌及１３例妇科非癌患者手术切除组织染色体１７ｐ１３．３的ＬＯＨ。结果１２例卵巢癌（包括１例交界性粘液性囊腺癌）和４例宫颈癌发生１７ｐ１３．３的ＬＯＨ,丢失频率分别为５０．０％和４４．４％。１３例非癌组织中,仅１例（７．７％）发生丢失,该例经病理证实为宫颈上皮内瘤变Ⅲ级,属癌前期病变（Ｐ＜０．０１）。结论染色体１７ｐ１３．３的ＬＯＨ可能与宫颈癌和卵巢癌的发生相关,检测１７ｐ１３．３的杂合性丢失将有助于深入了解卵巢癌和宫颈癌发生及发展的分子基础。     

人非小细胞肺癌中FHIT等位基因缺失和突变的研究

目的　探讨FHIT等位基因缺失、突变在肺癌发生、发展中的作用。方法　应用PCR SSCP和DNA序列分析方法对 3 5例人非小细胞肺癌和 4个肺癌细胞株中FHIT基因的 4个外显子 (外显子 3、4、5、8)和微卫星D3S13 0 0、D3S13 12、D3S13 13进行研究 ,并以远癌肺组织和 10例肺良性病变组织做对照。结果　在 3 5例肺癌中 ,2 2例肺癌发生了一个或两个以上的FHIT等位基因缺失 ,缺失率为 62 .86% ( 2 2 /3 5 )。在鳞癌中 ,FHIT等位基因缺失率 ( 88.2 4% ,15 /17)明显高于腺癌 ( 3 8.89% ,7/18) (P <0 .0 1) ;在吸烟患者中 ( 76.19% ,16/2 1)亦明显高于不吸烟患者 ( 4 2 .86% ,6/14 ) (P <0 .0 5 )。而FHIT等位基因缺失与肺癌的细胞分化程度、P TNM分期、原发肿瘤大小、部位、患者性别、年龄及有无转移均无明显关系 (P >0 .0 5 )。Lewis肺癌、A5 49细胞株亦有FHIT基因部分缺失。 4例肺癌组织具有微卫星灶D3S13 12点突变 ,经DNA序列分析显示均为D3S13 12微卫星灶基因的 87位点密码子发生了CT点突变。结论　FHIT基因异常主要以等位基因的缺失为主 ,而点突变发生率较低。FHIT等位基因缺失主要发生在肺鳞癌和吸烟患者中 ,且FHIT基因可能为烟草致肺癌的靶基因 ,其等位基因缺失可能是肺癌的早期分子事件。     

非小细胞肺癌微切割组织中3p杂合性丢失的研究

目的:探讨微切割非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中3p14.2和3p25等位基因的杂合性丢失(loss of heterozygosity,LOH)与NSCLC发生发展之间的相关性.方法:采用微切割技术和聚合酶链反应(PCR)结合二核苷酸(CA)n重复序列多态性方法,分析44例NSCLC石蜡切片中癌组织和非癌组织中3p14.2和3p25位点的杂合性丢失(LOH).结果:在44例NSCLC中29例组织标本检测3p14.2位点的LOH,其丢失率为65.5%(19/29),而另外15例患者中3p25的LOH的丢失率为40.0%(6/15).微切割的非癌组织均未见有3p的丢失,癌组织的3p LOH频率与肺癌组织学分类及TNM分期均无明显关系(P＞0.05).结论:NSCLC癌组织中普遍出现3p14.2及3p25杂合性丢失现象,从而提示该两处可能存在一个或多个与人体肺细胞癌变相关的抑癌基因.     

肺癌中ING1基因变化及其表达水平的研究

目的：检测肺癌中抑癌基因ING1微卫星杂合性缺失（LOH）及其主要蛋白产物p33^ING1b的表达情况，以探讨ING1基因改变-9肺癌发生发展的关系。方法：采用银染PCR—SSCP法检测肺癌组织ING1基因微卫星LOH发生的频率；应用组织芯片技术和免疫组化方法，检测肺癌p33^ING1b蛋白表达水平。结果：70例肺癌组织ING1基因4个微卫星位点的总杂合性缺失率为55．7％（39／70），并且越靠近ING1基因位点，发生率越高，但与临床病理参数无关。217例肺癌p33^ING1b蛋白的LOH发生率为47．0％（102／217），鳞状细胞癌高于腺癌、腺鳞癌和细支气管肺泡癌（P〈0．05），其余类型间差异无显著性（P〉0．05）；p33^ING1b的表达与其他临床病理参数不相关（P〉0．05），但与LOH发生频率相关（P〈0．05）。结论：p33^ING1b蛋白在肺癌组织中存在高频率的低表达或失表达，并且ING1基因的微卫星LOH发生频率也很高。推测LOH是该基因异常表达的重要原因，其结果可能导致该基因的下调和（或）蛋白质功能的失活，从而丧失其对细胞的生长抑制作用，促进了肿瘤的发生。     

Allelotype of non-small cell lung cancer

Loss of heterozygosity (LOH) studies have been used extensively to identify regions on chromosomes that may contain putative tumour suppressor genes. We have undertaken extensive allelotyping of 45 specimens of non-small cell lung cancer (NSCLC) using 92 polymorphic microsatellite markers on 39 chromosome arms. The most frequent allelic imbalances were found on chromosome arms 3p, 9p and 17p. Significant allelic imbalance was found on other chromosome arms including, 5q (21%), 8p (19%), 13q (24%) and 17q (18%). The LOH data on 3p was subdivided into the four chromosomal regions considered to contain putative tumour suppressor genes 3p25-p24 (10%), 3p21 (10%), 3p14 (25%) and 3p13-p12 (22%). The frequency of loss in the different regions on 9p were: 9pter-p23 (31%), 9p23-p22 (45%) and 9p21-cent (30%). LOH on 17p was separated into three regions: 17pter-p13 (9%), 17p13 (33%) and 17p13-cent (22%). No correlation was found between LOH on any of the chromosomal arms and any of the clinicopathological parameters such as pathology, level of differentiation, TNM staging or alcohol intake. Only one significant association was found between LOH and tumour types. A significant difference was found between LOH on 17q in adenocarcinomas and squamous cell carcinomas (p=0.037). The fractional allele loss (FAL) values for this group of 45 NSCLC gave a median value of 0.9 (range 0-0.45). No correlation was found between FAL and nodes at pathology (p>0.05) and between FAL and tumour grade (p>0.05). No correlation was found between p53 or ras mutations in these NSCLC specimens and their FAL values. Accumulated genetic damage, as provided by this allelotype analysis, provides a useful molecular parameter by which to assess NSCLC and may, in time, assist in the determination of the clinical behaviour and clinical outcome of these tumours.     

肺癌组织9p21区域杂合性缺失和p16基因异常的分析

目的：研究肺癌组织９ｐ２１杂合性缺失和ｐ１６基因的变化。方法：４９例癌组织和相应的１６例转移性肺门淋巴结ＲＮＡ进行ＲＴ－ＰＣＲ和ＳＳＣＰ分析，对其中３８例肺癌组织ＤＮＡ进行杂合性缺失分析（９不７１和ＩＦＮＡ两个位点）。结果：９ｐ２１区域杂合笥缺失频率：４７．３％（１８／３８）。Ｄ９Ｓ１７１和ＩＦＮＡ位点杂合性缺失频率分别为：２６．３％（１０／３８）和３６．８％（１４／３８）。原发灶和转移性林巴结果     

人非小细胞肺癌组织p63和p73的表达及意义

目的 :探讨p5 3家族新成员p6 3和p73在非小细胞肺癌 (NSCLC)中的表达及其临床意义。方法 :利用免疫组化S P法对 6 0例NSCLC和 7例正常肺组织检测p6 3和p73基因的蛋白表达。结果 :在NSCLC中 ,p6 3和p73蛋白表达的阳性率分别为 80 0 % (4 8/ 6 0 )、73 3% (4 4/ 6 0 ) ;与正常肺组织相比 ,p6 3和p73蛋白阳性表达的差异均有显著意义 ,P <0 0 5。p6 3蛋白表达与肺癌组织学类型 (P =0 0 0 0 3)和淋巴结转移有关 ,P =0 0 2 8;而与分化程度和临床分期无关 ,P >0 0 5。p73蛋白表达与肺癌组织学类型、淋巴结转移、分化程度和临床分期均无关 ,P >0 0 5。在肺癌中 ,p6 3蛋白和p73蛋白表达之间呈显著正相关 ,P =0 0 0 0 1。结论 :p6 3和p73高水平的表达协同促进肺癌的形成 ,p6 3是肺鳞癌恶性进展的一个标志物。     

Tobacco smoke-induced DNA damage and an early age of smoking initiation induce chromosome loss at 3p21 in lung cancer

The short arm of chromosome 3 is thought to harbor a novel oncogenic locus that is important in the genesis of lung cancer. The region at 3p21 is believed to contain a distinct locus that is sensitive to loss from the action of tobacco smoke carcinogens and has been reported to be specifically targeted for deletion in lung cancer. To investigate whether 3p21 alteration in lung cancer is associated with carcinogen exposure, PCR-based analysis was performed to detect loss of heterozygosity (LOH) on chromosome 3 at 3p21 in non-small cell lung carcinoma (NSCLC). We also measured instability at the BAT-26 locus, because the mismatch DNA repair gene, hMLH1, is found at 3p21. LOH at 3p21 was analyzed for association with the clinical features of NSCLC, p53 mutation status, polynuclear aromatic hydrocarbon-DNA adduct levels (measured using 32P-postlabeling) and carcinogen exposure information including cigarette smoking and asbestos exposure. Of 219 lung cancers, 150 cases (68.5%) were informative at the D3S1478 locus, and 44.2% of squamous cell carcinoma cases and 30.2% of adenocarcinoma cases showed 3p21 LOH. None of the cancers showed BAT-26 instability. The prevalence of 3p21 LOH was higher in both current and former smokers compared with never smokers and was higher in p53 mutated cases. Among squamous cell carcinoma cases, there was a strong association of increased 3p21 LOH with increasing polynuclear aromatic hydrocarbon-DNA adducts levels (P = 0.03), as well as an increased prevalence LOH with earlier age of smoking initiation (P = 0.02). Our results confirm that 3p21 LOH is strongly associated with measures of biologically effective dose of exposure to tobacco carcinogens. Our results also suggest that alterations of hMLH1 are not related to any of the reported associations, because there was no evidence of microsatellite instability. Finally, LOH in 3p21 may be an early molecular event in NSCLC, because it is significantly associated with a tendency to start smoking at a young age.     

Loss of heterozygosity is related to p53 mutations and smoking in lung cancer

Carcinogenesis results from an accumulation of several genetic alterations. Mutations in the p53 gene are frequent and occur at an early stage of lung carcinogenesis. Loss of multiple chromosomal regions is another genetic alteration frequently found in lung tumours. We have examined the association between p53 mutations, loss of heterozygosity (LOH) at frequently deleted loci in lung cancer, and tobacco exposure in 165 tumours from non-small cell lung cancer (NSCLC) patients. A highly significant association between p53 mutations and deletions on 3p, 5q, 9p, 11p and 17p was found. There was also a significant correlation between deletions at these loci. 86% of the tumours with concordant deletion in the 4 most involved loci (3p21, 5q11-13, 9p21 and 17p13) had p53 mutations as compared to only 8% of the tumours without deletions at the corresponding loci (P< 0.0001). Data were also examined in relation to smoking status of the patients and histology of the tumours. The frequency of deletions was significantly higher among smokers as compared to non-smokers. This difference was significant for the 3p21.3 (hMLH1 locus), 3p14.2 (FHIT locus), 5q11-13 (hMSH3 locus) and 9p21 (D9S157 locus). Tumours with deletions at the hMLH1 locus had higher levels of hydrophobic DNA adducts. Deletions were more common in squamous cell carcinomas than in adenocarcinomas. Covariate analysis revealed that histological type and p53 mutations were significant and independent parameters for predicting LOH status at several loci. In the pathogenesis of NSCLC exposure to tobacco carcinogens in addition to clonal selection may be the driving force in these alterations.     

Genetic alterrations of microsatellite markers at chromosome 17 in non-small cell lung cancer

Despite the high incidence and mortality, the exact molecular mechanism of the tumorigenesis and progression of lung cancer is still unclear. Alterations in oncogenes and onco-supressor genes have been described in lung cancer. Recent studies have started to define microsatellite instability (MI) and loss of heterozygosity (LOH) in genetic material of patients with lung cancer.[1] Microsatellite instability represents mutations of the short-tandem repeat sequences distributed within the geno…     

Frequent FHIT gene loss of heterozygosity in human papillomavirus-infected non-smoking female lung cancer in Taiwan

The fragile histidine triad (FHIT), located in chromosome region 3p14.2, had been reported to be a frequent allele with loss of heterozygosity (LOH) in smoking lung cancer and HPV-associated cervical cancer. Additionally, FHIT LOH may act as a tumor suppressor gene to involve in smoking-related lung tumorigenesis and HPV-related cervical tumorigenesis, respectively. In our previous report, a high prevalence of HPV 16/18 infection has been observed in non-smoking female lung cancer patients, and thus it was speculated that HPV 16/18 infection may increase the LOH frequency of FHIT in female cases to implicate in lung tumorigenesis. In this study, 157 lung cancer patients were enrolled and subjected to FHIT LOH analysis with three microsatellite markers. As expected, the frequency of FHIT LOH in males, smokers, and squamous cell carcinomas lung cancer patients was significantly higher than that of their corresponding counterpart (P=0.020 for gender, P<0.001 for smoking status, and P=0.038 for tumor type). Interestingly, a correlation between HPV 16 infection and FHIT LOH was observed in female lung cancer cases. To be more specifically, FHIT LOH frequency was remarkably increased from 18% (6 of 33) in HPV 16 non-infected female cases to 46% (11 of 24) in HPV 16 infected cases. The higher frequency of FHIT LOH observed in HPV 16-infected female lung tumors suggested that the involvement of HPV infection in lung tumorigenesis may, at least in part, be mediated through FHIT LOH.