CDKN2基因的变异在人前列腺增生症中发病机制的研究

目的：研究抑癌基因ＣＤＫＮ２的突变、缺失在人前列腺增生症（ＢＰＨ）病因中的作用机制。方法：用ＰＣＲ－银染ＳＳＣＰ技术分析了２０例正常前列腺组织和４１例ＢＰＨ中ＣＤＫＮ２基因纯合性缺失和突变的情况。结果：１３例ＢＰＨ有ＣＤＫＮ２基因缺失，总缺的率为３１．６％；正常前列腺组织中有１例出现ＣＤＫＮ２基因缺失，缺失率为５％（Ｐ〈０．０５）；无ＣＤＫＮ２基因的突变。结论：ＣＤＮＫ２基因的与ＢＰＨ的发病机制有     

MTS1基因在人BPH中突变缺失的初步研究

目的探讨ＭＴＳ１基因在人前列腺增生症（ＢＰＨ）发病机制中的作用和变异情况。方法用ＰＣＲ银染ＳＳＣＰ技术检测１８例正常前列腺和３８例ＢＰＨ中抑癌基因ＭＴＳ１各外显子的纯合性缺失和突变。结果１２例ＢＰＨ出现ＭＴＳ１基因缺失,总缺失率为３２％;正常前列腺组织中有１例出现ＭＴＳ１基因缺失,缺失率为６％,两者之间差异有显著性（Ｐ＜００５）;无１例有ＭＴＳ１基因的突变。结论ＢＰＨ的发病机制可能与ＭＴＳ１的纯合性缺失有关,未见有突变发生。应对ＭＴＳ１基因在人ＢＰＨ中的作用作更深入的研究     

BPH和前列腺腺癌中Livin的表达及其意义

目的：探讨凋亡抑制因子Livin在BPH和前列腺腺癌中的表达及其临床意义。方法：采用免疫组织化学SP法,用Livin单克隆抗体对10例BPH、30例前列腺腺癌和10例正常前列腺组织中Livin基因的表达进行检测,分析其在正常前列腺组织、BPH组织和前列腺腺癌组织中的表达状况,以及Livin基因的表达与前列腺腺癌病理学分级之间的关系。结果：Livin基因在10例正常前列腺组织中均不表达,而在10例BPH组织中有8例呈阳性表达（80％）,在30例前列腺腺癌中有21例呈阳性表达（70％）。Livin基因在正常组织中的阳性表达率与在BPH、前列腺腺癌中的阳性表达率差异有明显的统计学意义（P〈0．01）;Livin基因在BPH和前列腺腺癌中的阳性表达率差异无明显的统计学意义（P〉O．05）;Livin基因的表达和前列腺腺癌的分级无明显关系（P〉0．05）。结论：细胞凋亡抑制因子Livin在BPH和前列腺腺癌组织中的表达上调,提示Livin基因可能通过抑制细胞凋亡对BPH和前列腺腺癌的发生和发展起重要作用。检测Livin基因的表达对鉴定前列腺组织的良恶性方面无明显作用。     

CDKN2基因变异致前列腺增生症发病机制的研究

ＣＤＫＮ２基因位于人染色体９Ｐ２１～２２区，研究表明，其在各种恶性肿瘤中存在高频率的缺失、突变、甲基化和重排等变异。前列腺增生症（ＢＰＨ）是常见的老年男性疾病，其病因尚未阐明。为了探讨ＣＤＫＮ２基因与ＢＰＨ生物学行为之间的关系，我们采用ＳＳＣＰ技术分析了２０例正常前列腺组织和４１例ＢＰＨ组织中ＣＤＫＮ２基因纯合性缺失和突变情况。现将结果报告如下。１　材料与方法１．１　标本与试剂本组６１例，其中ＢＰＨ组织４１例，非前列腺增生组织（包括正常前列腺组织１０例，前列腺炎组织１０例）２０例，均来自湖北医…     

食管癌p16基因变异分析

目的 为了探讨Ｐ１６基因与食管癌发生、发展的关系。方法 应用聚合酶链反应、单链构象多态分析（ＰＣＲ－ＳＳＣＰ）方法检测３０例食管癌中Ｐ１６基因的纯合性缺失和突变情况，并分析Ｐ１６基因变异与食管癌的是关系。结果 ３０例食管癌患者中４例Ｐ１６基因纯合性缺失，２例存在突变。６例阳性样品与其它样品在年龄、饮酒史方面差异有显著性（Ｐ〈０．０１），与淋巴结转移、病理分期有差异（Ｐ〈０．０５），而与性别、肿瘤部     

人非小细胞肺癌P16基因突变研究

目的探讨ｐ１６基因缺失和突变与人非小细胞肺癌发生、发展的关系。方法应用多重ＰＣＲ、ＳＳＣＰ分析及ＤＮＡ序列分析技术，对４５例人非小细胞肺癌组织中ｐ１６基因外显子１和外显子２进行基因缺失和突变分析。结果４５例肺癌样品中共检出１９例ｐ１６基因缺失，总的缺失率为４２．２％（１９／４５）。其中纯合型缺失８例（１７．８％），半合型缺失１１例（２４．４％）；外显子１缺失７例（１５．６％），外显子２缺失１２例（２６．７％）；ｐ１６基因外显子２的突变检出率为３１．１％（１４／４５）；ＤＮＡ序列分析显示ｐ１６基因７２位密码子无义突变７例，７５位密码子错义突变７例。结论ｐ１６基因的缺失和突变可能与人非小细胞肺癌的发生、发展有关。     

乳癌中p16,c-erbB-2的表达及意义

探讨ｐ１６和ｃ－ｅｒｂＢ－２蛋白表达与乳癌临床病理因素及预后的关系。方法 采用免疫组化和ＰＣＲ－ＳＳＣＰ技术,分别检测５０例乳癌ｐ１６,ｃ－ｅｒｂＢ－２蛋白的表达和２０例乳癌ｐ１６基因的纯合性缺失及突变。结果 ５０例乳癌中,１７例（３４．０％）ｐ１６蛋白表达阳性,２４例（４８．０％）ｃ－ｅｒｂＢ－２蛋白表达阳性。２０例乳癌中无ｐ１６基因的纯合性缺失,１例ｐ１６基因外显子２有点突变。结果表明,ｐ１６     

前列腺组织中PTTG的表达及其意义

目的：研究垂体肿瘤转化基因（PTTG）在前列腺组织,尤其是前列腺癌组织中的表达及临床意义。方法：采用免疫组织化学二步法检测正常前列腺（8例）、BPH（16例）、无远处转移的前列腺癌（47例）以及有远处转移的前列腺癌（19例）组织中PTTG的表达情况。用Stata统计软件分别对BPH和前列腺癌组织中PTTG的表达情况与PSA和Gleason分级间的关系进行分析。结果：PTTG在正常前列腺和BPH组织中无表达或弱表达,而63．7％（42／66）的前列腺癌中有阳性或强阳性表达,在良恶性前列腺组织之间的表达差异有统计学意义（x2=25．2487,P=0．000）,无远处转移的前列腺癌和有远处转移的前列腺癌之间的表达差异无统计学意义（x2=0．3568,P=0．837）。PTTG在前列腺癌中的表达与PSA和Gleason分级之间无相关性。结论：PTTG在前列腺癌组织中高表达,可能在前列腺癌的发生发展过程中起重要作用。     

C-myc基因扩增、p16基因变异和乙型肝炎病毒DNA整合与肝细胞癌的相关性研究

目的 探讨C－myc基因扩增、p16基因变异和乙型肝炎病毒（HBV）DNA整合与肝细胞癌（HCC）的相关性。方法 应用差异聚合酶链反应（d-PCR)分析C－myc基因扩增,用聚合酶链反应（PCR）和单链构象多态（SSCP）分析p16基因变异,用PCR法检测整合型HBV－DNA。结论 （1）HCC组外周血、癌和癌旁肝组织中C－myc基因扩增阳性率分别为48％（14／29）、45％（13／29）和52％（15／29）,3者间差异无显著意义（x^2＝0．0008,P＞0．05,自由度ν＝2）,但明显高于肝硬化（LC）组外周血和LC组织阳性率[0(0/12)和8％（1／12）,P均小于0．05]。（2）29例癌组织中有3例p16基因纯合性缺失,未发现点突变。（3）正常肝、肝硬化和肝癌组织中整合型HBV－DNA阳性率分别为14／5（2／14）、67％（8／12）和97％（28／29）,3者间差异有显著意义（x^2＝29．4345,P＜0．01）。结论（1）HCC的发生与C－myc基因扩增密切相关;（2）HCC中p16基因的纯合性缺失发生频率为10％;（3）HBV－DNA整合与HCC的发生密切相关;（4）对外周血的检测能反映肝细胞（C－myc基因扩增和HBV－DNA整合情况。     

h-Spry2基因在部分人类肿瘤中的表达和突变研究

目的 探讨h-Spry2基因在136例肿瘤发生中的作用。方法 分别采用逆转录－聚合酶链式反应（RT－PCR）、聚合酶链式反应（PCR）、单链构象多态性分析（SSCP）和DNA测序技术检测h-Spry2基因在部分人类肿瘤及肿瘤细胞系中的表达、缺失和突变。结果 h-Spry2基因在71例癌肿瘤及瘤旁正常组织中均有表达，表达差异无显著性无定量分析，故无统计学处理。该基因在65例骨及软组织肿瘤中无纯合性缺失。在MKN45细胞系cDNA中存在15处同义突变；在DLD－1细胞系cDNA和基因组DNA中存在1处相同的同义突变。h-Spry2基因与果蝇Sprouty同源序列的扩增片段等长于用相同引物扩增的cDNA片段长度。结论 h-Spry2基因在上述肿瘤的发生中不起作用；h-Spry2基因与果蝇Sprouty同源序列中不含有内含子。     

CDKN2／p16^INK4a基因克隆,探针制备及其在肺癌基因诊断中的 …

OBJECTIVES: To clone CDKN2/p16(INK4a) gene, prepare its probe, and to study the change of CDKN2/p16(INK4a) gene in lung cancers. METHODS: Total RNA of normal lung tissue was extracted, CDKN2/p16(INK4a) gene cDNA synthesized, and CDKN2/p16(INK4a) gene recombinant vector, constructed. Southern blot was used to study CDKN2/p16(INK4a) gene in 46 cases of lung cancers, 3 cases of normal lung tissues, 6 cases of lung tissues near cancer, and 3 cases of lymph nodes with lung cancer metastasis. RESULTS: Cloned CDKN2/p16(INK4a) cDNA was proved by enzyme digestion and sequencing. Southern blot showed 4.3 kb band in normal lung tissues and lung tissues near cancers, and deletion of CDKN2/p16(INK4a) gene in cancer tissues and lymph nodes with lung cancer metastasis, with a deletion rate of 17.4% (8/46). CONCLUSION: CDKN2/p16(INK4a) gene may play a role to some extent in progression of lung cancers.     

乳腺癌细胞p16基因的研究

目的 探讨Ｐ１６基因在乳腺癌中的突变形式和频率以及有效的检测手段。方法 采用细胞２,反复贴壁法纯化人乳腺癌细胞、经多聚酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）分析技术。对１４例纯化乳腺癌细胞标本,１４例新鲜未纯化标本和２７例石蜡包埋标本及相应的癌旁乳腺组织标本的这６基因突变进行了对比性研究。结果 纯化的癌细胞组Ｐ１６基因突变率为４２．９％,而新鲜未纯化细胞组、石蜡包埋细胞组及癌旁乳腺细胞组的Ｐ     

CDKN2/p16基因存在状态与肺癌

目的　研究CDKN2 /p16基因缺失和蛋白丢失在肺癌发展中的作用。方法　采用免疫组化和改良标本处理的PCR技术 ,对 89例肺癌病人手术标本CDKN2 /p16基因第 1、2外显子纯合缺失和P16蛋白表达丢失情况进行分析对比研究。结果　CDKN2 /p16基因第 1或 (和 )第 2外显子总缺失率为2 5 8% (2 3/ 89) ,P16蛋白丢失率为 47 2 % (4 2 / 89) ,基因的缺失和蛋白的丢失集中发生于非小细胞肺癌(NSCLC) ,并与转移和分期有关。结论　CDKN2 /p16基因的异常可能是NSCLC区别于小细胞肺癌(SCLC)的特有的分子事件 ,其在NSCLC的发生发展中起一定的负调控作用     

Concordant loss of MTAP and p16/CDKN2A expression in gastroesophageal carcinogenesis: evidence of homozygous deletion in esophageal noninvasive precursor lesions and therapeutic implications

The gene that encodes methylthioadenosine phosphorylase (MTAP), an enzyme involved in adenine and methionine salvage pathways, is located on chromosome 9p21 telomeric to the p16INK4A/CDKN2A tumor suppressor gene. Inactivation of the p16INK4A/CDKN2A gene occurs by three different mechanisms: hypermethylation of the gene promoter, intragenic mutation coupled with loss of the second allele, and homozygous deletion. Immunohistochemical labeling for the p16INK4A/CDKN2A gene product parallels gene status but does not elucidate the mechanism of gene inactivation. Since the MTAP gene is often co-deleted with p16INK4A/CDKN2A, concurrent immunolabeling for both proteins can identify cases with homozygous p16INK4A/CDKN2A gene deletion. MTAP loss itself has therapeutic implications since it may confer selective sensitivity to inhibitors of de novo purine biosynthesis, such as L-alanosine. Twelve tissue microarrays were constructed from 92 cases of Barrett-associated adenocarcinomas and precursor lesions and 112 cases of gastric adenocarcinoma and precursor lesions comprising 1161 individual cores. Multiple cores were arrayed from any given case, and when available, included the entire histologic spectrum of intestinal metaplasia-dysplasia-carcinoma. Tissue microarrays were labeled with monoclonal antibodies against MTAP protein (clone 6.9, Salmedix, Inc) and p16 (clone 16P07, Neomarkers). Complete loss of labeling was considered negative, while any labeling (p16: nuclear; MTAP: cytoplasmic and nuclear) was considered positive. Loss of MTAP labeling occurred exclusively in conjunction with loss of p16 labeling, confirming that the previous findings from this group that concurrent loss of MTAP and p16 labeling is a surrogate marker of 9p21 homozygous deletions. Complete loss of MTAP and p16 was seen in 4 of 25 (16%) patients with Barrett's esophagus, 4 of 18 (22%) with low-grade dysplasia, 5 of 39 (13%) with high-grade dysplasia, 17 of 78 (22%) with invasive adenocarcinoma, and 8 of 36 (22%) of metastases. There were 7 cases of esophageal adenocarcinoma with loss of both MTAP and p16 for which precursor lesions were available. In 6 on these 7 cases (85%), the precursor lesion(s) had loss of both MTAP and p16. Lack of MTAP and p16 expression was seen in 11 of 106 (10%) cases of gastric adenocarcinoma. All metaplastic (30 biopsies from 20 cases) and dysplastic (15 biopsies from 13 cases) gastric tissues had both intact MTAP and p16INK4A/CDKN2A gene products. No precursor lesions were available from the gastric cancers that had loss of both MTAP and p16. Two benign gastric hyperplastic polyps also had intact p16 and MTAP. Concurrent MTAP and p16 loss detected by immunohistochemistry can serve as a convenient surrogate for p16INK4A/CDKN2A gene homozygous deletion in archival tissues. Inactivation of p16INK4A/CDKN2A by homozygous deletion appears to be an early event in Barrett carcinogenesis, occurring in noninvasive precursor lesions, including nondysplastic Barrett mucosa, in subsets of cases. In the absence of MTAP, cells depend exclusively on the de novo synthesis pathway for production of adenosine. This loss of MTAP during 9p21 homozygous deletion might be exploited therapeutically using de novo purine synthesis antimetabolites to treat a subset of invasive gastroesophageal adenocarcinomas and esophageal precursor lesions.     

抑癌基因p16和RB与肺癌早期诊断的临床实验研究

目的 研究肺癌组织中ｐ１６和ＲＢ基因失活的比率和失活的机制,探讨其与肺癌生物学特性及临床、病理和基因分型诊断的关系。方法 采用免疫组化、双重原位杂交、ＰＣＲ、ＰＣＲ－ＳＳＣＰ以及序列分析等方法,检测１０６例肺癌患者的癌组织和正常肺组织以及２３例肺良性疾病标本抑癌基因ｐ１６和ＲＢ的改变,并做对比研究。结果 肺癌细胞ｐ１６ＲＢ在蛋白和ｍＲＮＡ水平的表达明显低于正常肺组织和良性疾病肺组织,且与肺癌组织学类型、有无淋巴结转移以及临床病理分期密切相关,较早期肺癌（Ⅰ、Ⅱ期）病例即 较显著的抑癌基因０ｐ１６和ＲＢ的失活（３２．６％和２８．３％）;非小细胞肺 以ｐ１６基因失活为主（５０．１％）,小细胞肺癌以ＲＢ基因失活为主（８８．２％）,ｐ１６基因失活的主要机制有纯合缺失、甲基化和点突变。结论 抑癌基因ｐ１６和ＲＢ在肺癌发生     

原发性肝癌中CDKN2/P16基因突变的研究

目的 阐明ＣＤＫＮ２／Ｐ１６基因突变与原发性肝癌发生的关系。方法 采用ＰＣＲ－ＳＳＣＰ及ＰＣＲ产物直接银染测序技术，对３０例原发性肝癌组织中ＣＤＫＮ２／Ｐ１６基因第二外显子进行突变检测。结果 检出２例（２／３０）ＣＤＫＮ２基因突变，均是位于第１２４位密码子的错义突变。结论 ＣＤＫＮ２基因的突变虽不频发，但在原发性肝癌的发生中起了一定作用。     

Absence of p16 gene (CDKN2) deletions in microdissected primary breast carcinoma specimens

Background: The p16 gene (CDKN2), a tumor suppressor gene located on chromosome 9p21, has been demonstrated to be mutated or deleted with high frequency in a variety of tumor cell lines, including breast. While previous studies have not demonstratedCDKN2 mutations in primary breast carcinomas, it is possible that gene deletion in neoplastic DNA was masked by the presence of contaminating normal stromal DNA in breast carcinoma specimens. Methods: We investigated the incidence of homozygous deletion ofCDKN2 by analyzing 20 microdissected pure populations of primary breast carcinoma cells. Using polymerase chain reaction (PCR) techniques, the entire coding region and intervening introns ofCDKN2 were amplified. The PCR products were resolved by agarose gel electrophoresis and single-strand conformation polymorphism (SSCP) analysis. Results: We detected no deletions or mutations of the p16 gene. Conclusions: CDKN2 is not deleted with high frequency in primary breast carcinomas, and the p16 gene does not play a role in breast carcinogenesis via this mechanism. Presented at the 49th Annual Cancer Symposium of The Society of Surgical Oncology, Atlanta, Georgia, March 21–24, 1996.     

前列腺癌CD44及nm23-H1基因表达

目的　探讨在前列腺癌中CD44和nm2 3 H1基因表达的意义。　方法　应用免疫组织化学、银染单链长度构相多态性半定量逆转录聚合酶链式反应及免疫印迹杂交法分别检测 32例前列腺癌石蜡标本及 15例新鲜标本中CD44、nm2 3 H1基因的突变、表达情况。　结果　前列腺癌组织中nm2 3 H1蛋白及mRNA高水平表达 ,转移组nm2 3 H1蛋白表达强度高于非转移组。前列腺癌组织中存在nm2 3 H1基因突变 ,突变检出率为 13 3% (2 / 15 )。前列腺癌转移组中 ,CD44s蛋白表达水平下降。CD44smRNA在癌及非癌对照组织中全部表达 ,但 86 7% (13/ 15 )的癌组织中同时表达CD44v。　结论　nm2 3 H1基因的高表达和CD44v/CD44s基因的表达失衡可能在前列腺癌的恶性进展中共同发挥作用。