膀胱移行细胞癌3号染色体短臂上抑癌基因杂合性缺失的研究

目的 探讨位于3号染色体短臂的候选抑癌基因FHIT、hMLH和VHL在中国人膀胱移行细胞癌（TCC）发生中的可能作用。方法 选取6个位于上述基因内含子或与上述基因紧密连锁的微卫星多态标记对40例膀胱TCC组织进行杂合性缺失（LOH）分析。结果 位于FHIT基因内含子内的两个微卫星多态标记（D2S1234和D3S1300）中至少有一个为杂合子的杂合率为95．0％（38／40）,至少有一个发生LOH的频率为57．8％（22／38）。与hMLH1基因连锁的两处微卫星多态标记（D3S1561和D3S1612）中至少有一个为杂合子的杂合率为55．0％（22／40）,至少有一个发生LOH的频率为59．1％（13／22）。与VHL基因连锁的两个微卫星多态标记（D3S1038和D3S1284）中至少有一个为杂合子的杂合率为90．0％（36／40）,至少有一个发生LOH的频率为47．2％（17／36）。在所检测微卫星多态标记中仅D3S1284的LOH与膀胱TCC的病理分期正相关（P＜0．01）,余微卫星多态标记的LOH与膀胱移行细胞癌病理分级及病理分期均不相关（P＞0．05）。结论 上述3个基因在膀胱TCC中存在高频率LOH,提示它们可能在膀胱TCC的发生发展中起重要作用。FHIT基因和hMLH1基因的缺失作为分子标志有可能为膀胱TCC的早期诊断提供新的途径和依据,而VHL基因的缺失可能是膀胱TCC发生的晚期事件。     

食管鳞癌组织中hMLH1基因微卫星变异检测

目的:探讨食管鳞癌及不典型增生组织中hMLH1基因微卫星变异。方法:采用PCR-变性聚丙烯酰胺凝胶电泳-银染技术,对40例食管鳞癌、40例正常及26例不典型增生组织中hMLH1基因所在区域的3个微卫星位点D3S1561、D3S1289及D3S1448进行检测,分析其微卫星不稳定(MSI)及杂合性缺失(LOH)状况。结果:在不典型增生组织中三个位点的MSI总阳性率为65.0%,高于在癌组织的30.0％(P<0.05);前者的LOH总阳性率为7.6％,与后者的7.5％比较差异无统计学意义(P>0.05)。癌组织的MSI及LOH与不典型增生组织的MSI及LOH的r分别为0.623和1(P均<0.05)。结论:错配修复缺陷参与了食管鳞癌的发生过程,且是早期分子事件;在食管鳞癌发生发展过程中,hMLH1基因的LOH并非一个常见和重要的事件。     

3p、9p微卫星DNA异常与肺癌的早期诊断

目的研究3p、9p微卫星DNA异常在肺癌早期诊断中的价值。方法应用PCR-银染法从外周血检测105例原发性肺癌及9例肺部良性疾病3p14、3p21、9p21上的3个微卫星位点（D3S1228、D3S1029、D9S171）的异常表现。结果肺癌患者血清中DNA含量多于良性病变患者。肺癌组各微卫星位点的微卫星不稳症（MSI）或微卫星杂合性缺失（LOH）异常的阳性率为43％。50％,以D3S1029最高,达49．6％,3个位点中至少1个位点出现微卫星异常者占76．2％．其中45．7％呈多位点改变,同肺良性病变组比较有显著性差异（P〈0．05）。肺癌组中各微卫星位点的异常表现与肺癌临床分期和临床病理类型之间无显著相关性沪〉0．05）。结论3p、9p微卫星DNA异常和肺癌分期无关,可以作为一项肺癌早期基因检测的新途径。不同微卫星位点出现异常表现的具体形式有所不同,多位点联合检测可以提高诊断的敏感性和特异性。     

急性淋巴细胞白血病患者微卫星DNA杂合性缺失检测

目的：探讨微卫星DNA杂合性缺失与急性淋细胞白血病（ALL）的相关性。方法：ALL患者72例，其中T－ALL9例，B－ALL55例，慢性粒细胞性白血病（慢粒）急淋为8例，应用PCR－变性聚丙烯酰胺凝胶电泳－银染技术，检测ATM基因D11S2179位点、BRCA2基因D13S269点位点和D13S25在B－ALL、T－ALL和慢粒急淋变患者的微卫星杂合性缺失（LOH）。结果：72例ALL患者中，3位点总LOH频率为26．4％。D11S2179、D13S260和D13S53位点LOH的发生频率分别为15．3％、12．5％和5．5％。B－ALL患者3个位点平均LOH发生率为12．7％。在D11S2179和D11S2602位点同时发生LOH频率为9．1％。9例T－ALL患者未发现LOH。8例慢性急淋变患者只在D11S2179位点检测到3例（37．1％），其他2位点未异常。结论：ATM基因、BRCA2基因帮D13S25位点微卫星的LOH与B－ALL的发病存在相关性；而ATM基因遗传不稳定性可能参与慢粒急淋变过程。     

T细胞非霍奇金淋巴瘤微卫星不稳定性及hMSH2、hMLH1蛋白表达研究

目的：探讨微卫星不稳定性（Microsatellite instability,MSI）及DNA错配修复基因系统（Mismatch repair gene system,MMR）功能缺陷在T细胞非霍奇金淋巴瘤（Non-Hodgkins lymphoma,NHL）发病机制中的意义。方法：对38例T细胞NHL采用PCR扩增,变性聚丙烯酰胺凝胶垂直电泳,银染方法检测17号染色体p53基因周围4个微卫星位点多态标志D17S945、D17S938、D17S947、D17S926的MSI,并运用免疫组织化学SP法检测肿瘤细胞hMsH2、hMLH1蛋白表达情况。结果：38例T细胞NHL中MSI的阳性率为23．68％（9／38）,LOH为7．89％（3／38）。各位点MSI、LOH频率介于2．86％～18．42％和0．00％～5．26％。各位点MSI阳性率差异无统计学意义（P〉0．05）;肿瘤组织中,hMSH2、hMLH1蛋白表达缺失率分别为52．63％（20／38）、42．11％（16／38）。结论：T细胞NHL中存在MSI和MMR蛋白表达缺陷。     

胶质母细胞瘤8号、22号染色体等位基因杂合性丢失的研究

目的：寻找胶质母细胞瘤（GBM）8号，22号染色体上可能存在肿瘤抑制基因的杂合性丢失（LOH）区域。为发展和定位肿瘤抑制基因（TSG）提供线索和依据。方法：应用聚合酶链反应（PCR）方法，采用荧光标记的引物和377型DNA序列自动分析仪，分别分析了21例GBM8号，22号染色体上14个，7个微卫星多态性标记的LOH。结果：本组病例8号染色体LOH率为23．8％，在16．3％能提供信息位点检测到LOH，8P的LOH率明显高于8q，分别为23．8％，14．3％，8q上所有位点的LOH率都在15％以上，以8p22-23上D8S550和D8S549的LOH率较高，分别是27．8％，30．0％，染色体22q的LOH率为47．6％，在30．0％可提供信息位点存在LOH，以22q13.2-13.3上D22S274的LOH率最高（41．2％），结论：8号，22号染色体等位基因杂合性丢失可能在GBM的分子发病机制中发挥着重要作用。在8p22-23上D8S550和D8S549位点间区域以及22q13.2-13.3上D22S274位点所在区域可能存在多个与GBM相关的肿瘤抑制基因。     

FHIT基因与宫颈癌的相关性研究

目的：探讨FHIT基因在宫颈癌发生中的作用及与宫颈癌的临床分期和组织分化程度间的关系。方法：选择FHIT基因的2个微卫星多态标记对42例宫颈癌进行杂合性丢失（LOH）和微卫星不稳定性（MI）分析。结果：在D3S1234和D3S1300座位上，LOH频率分别为52．5％（21／40），70．8％（17／24），MI频率分别为22．5％（9／40）、20．83％（5／24）。FHIT基因改变与宫颈癌的临床分期和组织分化程度无关。结论：FHIT基因参与了宫颈癌的发生。对于宫颈癌的诊断和筛查可能具有实用价值。     

人肝癌p57^kip2基因表达缺失与杂合性缺失的关系研究

目的通过对p57^kip2 mRNA表达水平下降的人体肝癌进行杂合性缺失研究,探讨肝癌发生有关基因调控的分子机制．方法对人肝细胞肝癌及癌周肝组织各30例,正常肝组织对照20例,共80例标本,采用原位杂交检测p57^kip2 mRNA的表达,并运用PCR-聚丙烯酰胺凝胶电泳-银染法和基因分型对30例肝癌组织及其癌周肝硬化组织p57^kip2基因所在染色体区域的10个微卫星位点（D11S1397,D11S1318,D11S4046,D11S922,TH01,D11S2359,D11S1760,D11S1338,D11S569和D11S1397）进行杂合性缺失检测,同时分析了p57^kip2 mRNA表达与杂合性缺失之间的关系．结果原位分子杂交检测正常肝组织未见p57^kip2 mRNA表达,癌周肝硬化组与肝癌组阳性表达率均为26．7％（8/30）．杂合性缺失检测结果显示在10个微卫星位点中,仅有TH01（7／30,23．3％）、D11S2359（5／30,16．6％）和D11S569（1／30,3．3％）3个位点有杂合性缺失;p57^kip2 mRNA和蛋白表达缺失主要与TH01位点相关．结论p57^kip2 mRNA和蛋白表达异常提示其可能在原发性肝癌的发生发展中起重要作用．与p57^kip2基因同一区域的10个微卫星位点不是p57^kip2基因LOH和MSI的频发位点,但p57^kip2 mRNA表达缺失与TH01位点的杂合性缺失相关．     

散发性结直肠癌染色体10q23～24区域杂合缺失分析

目的抑癌基因的杂合缺失（LOH）被认为是结直肠癌形成的通路之一,本实验拟通过对染色体10q23～24区的LOH分析,发现高频杂合缺失区域并筛查与结直肠癌相关的抑癌基因。方法7个荧光标记的微卫星引物（围绕D10S185位点）与83例结直肠癌的肿瘤和正常组织进行聚合酶链反应（PCR）反应。产物在ABI Prism377自动荧光测序仪进行电泳,以GeneSean3．1和Genotyper2．1软件进行扫描以及杂合缺失分析LOH。与临床病理因素之间的关系比较采用X^2检验。结果7个位点平均杂合缺失率为36．11％,以D10S583位点最高,达54．84％;最低是D10S205,21．3％。发现两个高频杂合缺失区域：一个在D10S583和D10S185之间,大约0．9cM（10q23．33）的距离;另一个在D10S1709和D10S1265位点之间,大约1．5cM（10q24．2～24．31）的距离。D10S1265位点的杂合缺失与Dukes分期显著相关,其余位点与临床病理因素均无显著相关。结论在散发性结直肠癌10q23-24发现了两个高频LOH区域：10q23．33和10q24．2～24．31。除对磷脂酶同族蛋白（PTEN）基因外,10q上可能存在与散发性结直肠癌相关的其他抑癌基因。     

FHIT基因与脊索瘤的相关性研究

目的：探讨脆性组氨酸三联体（fragile histidine triad,FHIT)基因在脊索瘤发生中的作用。方法：选择FHIT基因内3个微卫星移态标记对18例脊索瘤进行杂合性丢失（loss of hetorozygosity,LOH)和微卫星不稳定性分析（microsatellite instability,MI)。结果：在18例脊索瘤标本中，LOH的总发生率为44．4％，其中在D3S 1300，D3S4130和D3S1234的LOH发生率为37．5％、26．7％、23．1％。MI总发生主为33．3％，3个位点的MI发生率分别为25％、13．3％、15．4％，微卫星多态分析表明，FHIT基因在脊索瘤中既存在杂合性丢失，又存在微卫星不稳定现象。结论：FHIT基因参与了脊索瘤的发生，可能是脊索瘤候选抑癌基因之一。     

子宫内膜癌患者FHIT、SLIT2、EDNRB基因杂合性丢失

目的：探讨子宫内膜癌患者FHIT、SLIT2、EDNRB基因3个微卫星位点D3S1287、 D4S1593、D13S160杂合性丢失（loss of heterozygosity，LOH），以确定侯选的抑癌基因。方法：应用PCR-变性 PAGE-银染方法分别对35例子宫内膜癌患者癌组织及相对应的正常子宫内膜组织在FHIT、SLIT2、EDNRB基因3个微卫星位点D3S1287、D4S1593、D13S160 行LOH检测。结果：LOH总检出率为54.3%，D3S1287、D4S1593、D13S160位点分别为34.5%、20.5%、19.3%。FHIT、SLIT2、EDNRB基因的3个微卫星位点发生LOH率与子宫内膜癌手术-病理分期无明显相关性。结论：子宫内膜癌患者肿瘤组织在FHIT、SLIT2、及EDNRB基因的微卫星位点D3S1287、D4S1593、D13S160 均有LOH。FHIT、SLIT2及EDNRB基因为抑癌基因，其失活可能与子宫内膜癌的发生有关。     

胃癌患者6号染色体长臂遗传不稳定性

目的研究中国人胃癌6号染色体长臂微卫星不稳定性（microsatellite instability，MSI）和杂合性缺失（loss of heterozygosity，LOH）状况及与临床病理特征之间的联系。方法采用PCR-SSLP-银染方法对27例胃癌组织及其相应正常组织6号染色体长臂（6q）不同位置的4个位点进行MSI和LOH检测。结果27例信息个体中16例检测到一个或多个位点MSI，平均MSI频率59．2％；11例不具MSI的信息个体中7例存在1个或更多位点LOH，平均频率63．6％。MSI和LOH频发位点均为D6S434（6q16．3-q21）和D6S404（6q16．3-q23）。MSI和LOH在高分化胃癌和低分化胃癌各个时期均有发生．但高分化胃癌中高水平MSI（MSI-H）发生有增高趋势。结论MSI和LOH发生率与年龄、性别、组织分化程度、病理分期及发病部位等临床指标虽然无明显相关，但6q等位基因缺失关键区域与其它国家、地区相似，进一步证实6q上该区可能存在与胃癌相关的肿瘤抑制基因。     

散发性结直肠癌3号染色体等位基因杂合缺失

目的 了解散发性结直肠癌（SCRC）3号染色体等位基因杂合缺失（LOH）发生情况。探讨其与临床病理特征间的关系,并对3号染色体上可能的SCRC相关基因进行初步定位。方法 用覆盖3号染色体的13个微卫星标记对83例散发性结直肠癌进行LOH扫描分析。结果 3号染色体至少有两个位点发生LOH者占39%（29/74）,3p所选4个位点中至少有一个发生LOH者占37%（27/74）,3q所选9个位点中至少有一个发生LOH者占53%（39/74）。整条染色体上以D3S1300（3q14.2)位点LOH率最高,达54%（23/43）。3qLOH在远端结直肠癌较近端高发,3q及D3S1300LOH阳性肿瘤多表现为浸润型生长和局部侵犯,并多发于大于50岁的老年患者。结论 3号染色体存在散在分布及区域性高频等位基因LOH,并与SCRC临床病理资料相关,提示其上SCRC相关基因的存在。高频LOH位点D3S1300的发现,提示3p14.2附近区域的FHIT基因可能作为肿瘤抑制基因在结直肠癌的发生发展中发挥作用。     

中国人胃癌染色体11p15.5处杂合性缺失分析

目的　通过对中国人胃癌染色体 11p15 .5处杂合性缺失的研究 ,了解中国人群中胃癌病人在此区域杂合性缺失的情况。方法　从 66例胃癌病人的石蜡包埋的手术病理标本中 ,提取肿瘤及相应正常组织的DNA ,用荧光标记的方法选取 11p15 .5处的微卫星DNA标记进行PCR扩增 ,再将PCR产物进行变性聚丙烯酰胺凝胶电泳 ,电泳后行杂合性缺失分析。结果　 11/ 3 6( 3 0 .6% )的病人在D11S13 18处存在杂合性缺失 ;而D11S40 46位点处 ,13 / 60 ( 2 1.7% )的病例存在杂合性缺失 ;2 4/ 61( 3 9.3 % )的病例在 11p15 .5处至少有 1个位点存在杂合性缺失。杂合性缺失的频率与肿瘤病人的年龄、性别、肿瘤大小、肿瘤部位及淋巴结是否转移均无关。结论　高频杂合性缺失的区域可能有抑癌基因的存在。这将有助于阐明胃癌发生发展的分子机制 ,进而为对胃癌进行风险预测、基因诊断以及基因治疗提供可能     

Screening of tumor suppressor genes on lq31.1-32.1 in Chinese patients with sporadic colorectal cancer

Background As a model for both multistep and multipathway carcinogenesis, colorectal neoplastic progression provides paradigms for researching both oncogenes and tumor suppressor genes (TSGs). However, the mechanism of colorectal cancer (CRC) is not completely understood, and many genes may be involved in the colorectal carcinogenesis. The purpose of this study was to screen for the potential TSGs on chromosome lq31.1-32.1 in Chinese patients with sporadic colorectal cancer, to explore whether colorectal cancer in the Chinese population has unique genetic alterations and determine whether other putative TSGs exist and contribute to colon carcinogenesis. Methods Six polymorphic microsatellite markers, at a density of approximately one marker in every 1.6 cM, were chosen for refined loss of heterozygosity (LOH) mapping of lq31.1-32.1. Eighty-three colorectal cancer patients' tumor and normal DNA were analyzed via polymerase chain reaction (PCR) for these microsatellite markers. PCR products were eletrophoresed on an ABI 377 DNA sequencer. Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. On the basis of refined LOH mapping results, we undertook a microarray-based expression screening to identify tumor association genes in 19 of the CRC cases. Results The average LOH frequency of lq31.1-32.1 was 24.41%, with the highest frequency of 36.73% (18/49) at D1S2622, and the lowest of 16.42% (11/67) at D1S412. A minimal region of frequent deletion was located within a 2 cM genomic segment at D1S413-D1S2622. There was no significant association between LOH of any marker in the studied regions and the clinicopathological data (patient sex, age, tumor size, growth pattern, or Dukes stage). On the basis of refined mapping results, we chose 25 genes located in the D1S413-D1S2622 (lq31.3-32.1) region and presented a microarray-based high throughput screening approach in 19 sporadic CRC cases to identify candidate CRC related tumor suppressor genes. This study found 4 significantly down-expressed genes, including CSRP1, LMOD1, PPP1R12B and CFHL3. There was no significant association between expression levels of CFHL3, CSRP1, LMOD1, PPPIR12B and the clinicopathological data. By database searching, CSRP1 was hypothesized to be a colorectal cancer related tumor suppressor gene. Conclusions Through detailed deletion mapping, we found that the lq31.3-32.1 region might harbor one or more colorectal cancer related tumor suppressor gene(s). And by microarray-based high-throughput screening of candidate genes located in this region and by subsequent database searching, we present the first evidence that CSRP1 might be involved in the progression of CRC.     

人肝细胞癌4号染色体高频率杂合子丢失的研究

目的 为克了生新的肝细胞（HCC）相关肿瘤抑制基因的供位点依据。方法 使用4号染色体的12个微卫星DNA多态性标志,分析了48例原发性HCC的杂合子丢失（LOH）。结果 44％（21／48）和63％3（30／48）的肿瘤组织中至少有1个位点的LOH分别发生在4p和4q。丢失图排列鉴定了两个独立的共同丢失片段（CDR）。第1个定位在4q22-q25的CDR位于D4S392和D4S1625位点之间;第2个定位在4q27-q31的CDR位于D4S1625和D4S1652位点之间。结论 至少有两个肿瘤抑制位点存在于4q.     

Screening of tumor suppressor genes on 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer

Background As a model for both multistep and multipathway carcinogenesis, colorectal neoplastic progression provides paradigms for researching both oncogenes and tumor suppressor genes （TSGs）. However, the mechanism of colorectal cancer （CRC） is not completely understood, and many genes may be involved in the colorectal carcinogenesis. The purpose of this study was to screen for the potential TSGs on chromosome 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer, to explore whether colorectal cancer in the Chinese population has unique genetic alterations and determine whether other putative TSGs exist and contribute to colon carcinogenesis. Methods Six polymorphic microsatellite markers, at a density of approximately one marker in every 1.6 cM, were chosen for refined loss of heterozygosity （LOH） mapping of 1q31.1-32.1. Eighty-three colorectal cancer patients＇ tumor and normal DNA were analyzed via polymerase chain reaction （PCR） for these microsatellite markers. PCR products were eletrophoresed on an ABI 377 DNA sequencer. Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. On the basis of refined LOH mapping results, we undertook a microarray-based expression screening to identify tumor association genes in 19 of the CRC cases. Results The average LOH frequency of 1q31.1-32.1 was 24.41%, with the highest frequency of 36.73% （18/49） at D1S2622, and the lowest of 16.42% （11/67） at D1S412. A minimal region of frequent deletion was located within a 2 cM genomic segment at D1S413-D1S2622. There was no significant association between LOH of any marker in the studied regions and the clinicopathological data （patient sex, age, tumor size, growth pattern, or Dukes stage）. On the basis of refined mapping results, we chose 25 genes located in the D1S413-D1S2622 （1q31.3-32.1） region and presented a microarray-based high throughput screening approach in 19 sporadic CRC cases to identify candidate CRC related tumor suppressor genes. This study found 4     

Deletion of chromosomes 9p and 17 associated with abnormal expression of p53, p16/MTS1 and p15/MTS2 gene protein in hepatocellular carcinomas

Ｈｅｐａｔｏｃｅｌｌｕｌａｒｃａｒｃｉｎｏｍａ (ＨＣＣ)ｒａｎｋｓｔｈｉｒｄａｍｏｎｇｔｈｅｃａｕｓｅｓｏｆｃａｎｃｅｒｍｏｒｔａｌｉｔｙｉｎＣｈｉｎａ ,ａｎｄａｂｏｕｔ4 2 5%ｏｆｔｈｅｗｏｒｌｄ’ｓｎｅｗｃａｓｅｓｏｆＨＣＣｅａｃｈｙｅａｒｏｃｃｕｒｉｎＣｈｉｎａ 1ＥｐｉｄｅｍｉｏｌｏｇｉｃｓｔｕｄｉｅｓｐｒｏｖｉｄｅｅｖｉｄｅｎｃｅｔｈａｔｉｎｆｅｃｔｉｏｎｗｉｔｈｈｅｐａｔｉｔｉｓＢ (ＨＢＶ )ａｎｄ/ｏｒｈｅｐａｔｉｔｉｓＣ (ＨＣＶ ) ,ａｎｄｉｎｇｅｓｔｉｏｎｏｆａｆｌａｔｏｘｉｎＢｃｏｎ…     

Detailed deletion mapping of loss of heterozygosity on 9p13-23 in laryngeal squamous cell carcinoma by microsatellite analysis

Background This study was designed to investigate the hot spots of microsatellite loss of heterozygosity (LOH) on 9p13-23 in laryngeal squamous cell carcinoma and to find out the correlation between the incidence of microsatellite LOH and the clinicopathological parameters.Methods Tumor tissues were obtained from paraffin embedded sections with microdissection. Genomic DNA was extracted from tumor tissues and peripheral blood lymphocytes with the phenol-chloroform. Polymerase chain reaction (PCR) amplification and denaturing gel electrophoresis were carried out in a set of 42 squamous cell carcinoma (SCC) of larynx and corresponding peripheral blood lymphocytes using 13 highly polymorphic microsatellite markers on 9p13-23. The correlation was analyzed between microsatellite LOH at the high frequency on 9p13-23 and clinicopathological parameters in the patients with squamous cell carcinoma of larynx.Results Of the 42 laryngeal cancers, 41 (97.6%) showed LOH in at least one of the microsatellite markers tested on 9p13-23. The most frequently deleted marker was D9S162 in 17 of the 19 (89.5%) informative samples. The marker D9S171, which is located on 9p21, had LOH detected in 12 of the 15 informative cases (80.0%). LOH at the D9S1748 marker (closest to the p16 gene locus) was detected in 18 of the 36 informative cases (50.0%). Allelic deletion mapping revealed two minimal regions of LOH encompassing markers D9S161-D9S171 on 9p21 and IFNA-D9S162 on 9p22-23. Multiple LOH (≥4) on 9p21-23 was found more frequently in the patients under 60 years, with supraglottic SCC or cervical lymph node metastasis than those over 60 years, with glottic SCC or without cervical lymph node metastasis (P&lt;0.01 or 0.01, 0.05, respectively). On the contrary, there was no correlation between T stages or pathologic classification and the frequency of LOH on 9p21-23 in 42 SCC of Larynx.Conclusions These findings imply the presence of at least two putative tumor suppressor genes on 9p13-23 in laryngeal SCC. Multiple genetic alterations are probably implicated in supraglottic SCC with cervical lymph node metastasis in younger patients.