比较基因组杂交技术检测肝细胞癌染色体异常及其临床意义

目的探讨肝细胞癌染色体异常及其临床意义。方法运用比较基因组杂交技术检测25例肝细胞癌染色体DNA异常情况,并与临床指标作相关分析。结果 25例肝癌均存在不同程度的染色体DNA拷贝数的扩增或缺失,较为常见的染色体DNA异常是+1q(72%)、+1p(64%)、+2q(48%)、+2p(48%)、+5q(48%)、+Xq(48%)、+7q(44%)、-4q(48%)、-16p(48%)、-8p(40%)、-17p(36%);相关分析显示+17p、+18p、-8p、-13q、-11q、-8q染色体异常事件与临床指标部分相关。结论肝细胞癌存在明显的染色体异常,部分染色体异常事件是非随机性的,可能与肝癌的发生、发展有关,并与肿瘤的生物学行为和预后相关。     

成套荧光原位杂交探针在慢性淋巴细胞白血病染色体异常检测中的应用

目的 探讨成套探针荧光原位杂交（FISH）技术在慢性淋巴细胞白血病（CLL）患者染色体异常检测中的应用,并初步分析染色体异常与其他预后指标的相关性及其临床意义.方法 对21例初诊CLL患者同时应用序列特异性探针D13S25（13q14.3）、RB1（13q14）、ATM（11q22.3）、p53（17p13.1）及着丝粒探针CSP12（12p11.1-12q11.1）进行间期FISH检测,分析患者染色体异常的发生率,同时分析染色体异常与CLL患者年龄、性别、Binet分期、CD38表达及乳酸脱氢酶（LDH）间的相关性,并对生存情况进行初步观察.结果 21例CLL患者中,13例发现有染色体异常（61.90％）,其中11例为单条染色体异常,1例为2条染色体异常,1例为3条染色体的复杂异常.按染色体异常发生率的高低依次为13q14-9例（42.86％）（其中D13S25单独缺失7例,D13S25和RB1同时缺失2例）,＋123例（14.29％）,11q22-（ATM缺失）2例（9.52％）,17p13-（p53缺失）2例（9.52％）.染色体异常与患者年龄、性别、Binet分期、CD38表达以及LDH水平未发现相关性.结论 成套探针FISH技术能够快速、敏感、准确地检测CLL患者的染色体异常.     

广西地区乙肝病毒／黄曲霉毒素B1双暴露相关性肝细胞性肝癌微阵列比较基因组学的研究

目的研究乙肝病毒／黄曲霉毒素B1双暴露相关性肝细胞性肝癌（hepatocellularcarcinoma,HCC）染色体遗传学畸变的特点。方法将32例手术切除经病理证实为HCC的癌组织,按照乙肝病毒与黄曲霉毒素的暴露情况分为4个亚组：A组为HBV（＋）／AFB,（＋）10例;B组为HBV（＋）／AFB1（-）10例;C组为HBV（-）／AFB1（＋）6例;D组为HBV（-）／AFB1（-）6例。应用微阵列比较基因组杂交技术（ArrayCGH）检测分析其22对染色体DNA拷贝数的变化。结果32例HCC样本中,共发现573个染色体畸变区段（chromosomalaberrations,CNAs）。其中1q、4p、5p、6p、7p、8q、10p、17q、20p、20q和X主要表现为扩增区段;1p、2q、4q、8p、9p、10q、11q、13q、14q、16p、16q、17p、19p、19q、21q、22q和Y主要表现为缺失区段。同时,共检测出25个染色体发生高频畸变的区段（recurrentlyalteredregions,RARs）,其中lq21．1-q44、5p13．2-p15．3、6p12．1-p25．2、7q11．2-q35、8q11．2-q24．3、17q12-q25．2、18q12．3-q22．3和x为高频率扩增区段,而lp31．1-p36．2、2q23．2-q37．2、4q12-q35．2、6q14．1-q26、8p12-p23．2、9p21．1-p24．2、10q21.3-q26．2、13q12．1-q21．1、14q21．3-q32．2、16p12．1-p13．2、16q12．1-q24．1、17p12-p1313、19p13．1-p13．3、19q13．2-q13．4、21q21．3-q22．2、22q11．2-q13．2和Y染色体为高频缺失区段。8p12-p23．2缺失的发生率在进展期HCC（TNM分期为Ⅲ～Ⅳ期）中明显高于早期HCC（TNM分期为I～Ⅱ期）（仁0．038）。4q12-q35．2、13q12．1-q21．1的缺失及7q21．1-q35的扩增发生率在A组中最高。Cox模型分析结果示：在单因素分析中AFP水平、肿瘤大小、TNM分期、BCLC分期、侵袭与转移的发生、8p12-p23．2的缺失以及19p13．1-p13．3的缺失等为影响患者无瘤生存时间的危险因素（P〈0．05）.而在多因素分析中AFP水平、TNM分期以及8p12-p23．2的缺失等为影响患者无瘤生存时间的危险因素（P〈0．05o结论广西地区HCC染色体遗传学改变具有多样性,其中染色体19p13．1．p13-3的高频缺失可能为广西地区HCC特有的分子生物学特征之一。染色体8p12-p23．2的缺失可能为HCC的晚期事件,并且与患者的不良预后有关。染色体4q12-q35．2、13q12．1-q21．1缺失及7q21．1-q35扩增可能与HBV／AFB。双因素的协同致癌作用有关。     

大肠癌染色体组DNA拷贝数异常的临床病理学意义

目的分析大肠癌染色体组DNA拷贝数异常区域与患者临床病理特征的关系。方法应用比较基因组杂交技术（CGH）检测73例大肠癌患者的标本。结果大肠癌染色体8p12-pter丢失和8q23-qter扩增与淋巴结转移显著相关;而8q23-qter扩增和18q12-qter丢失与远处器官转移和（或）术后复发显著相关;8q23-qter扩增、8p12-pter和18q12-qter丢失,与大肠癌患者不良预后密切相关。Cox多因素分析表明,18q12-qter丢失在大肠癌是一个独立的不良预后生物学指标。结论采用CGH法检测染色体组DNA拷贝数异常能预测大肠癌患者的预后,并为临床治疗提供有意义的信息。     

肝母细胞瘤基因组不平衡性与染色体变异

目的建立稳定的比较基因组杂交（CGH）技术,探讨肝母细胞瘤（HB）染色体1p36杂合性缺失（LOH）的特点。方法应用CGH检测30例HBDNA的丢失或扩增;应用聚合酶链反应-简单重复序列多态性（SSLP）方法,对30例HB中染色体1p36上6个微卫星的杂合子丢失进行检测。结果每例HB细胞染色体均有不同程度的变异,HB常见增益的染色体区域是1q、2q、2p、8q、8p、12q和22q,常见丢失的染色体区域为1P、4q、4p、16q、17p和18q。30例HB中,1号染色体上6个基因座发生LOH的总频率为63．3％（19／30）,其中D1S199为最高（66．7％）,其次为D1S450（46．7％）。结论HB存在多条染色体DNA拷贝数扩增或丢失的区域;HB在染色体1p36上存在广泛的LOH;染色体变异引起相应瘤基因扩增和抑癌基因的丢失可能参与了HB的发生、发展。     

肝细胞癌临床病理学特征与p53蛋白表达的关系

应用免疫组织化学技术检测29例肝细胞癌（肝癌）和23例肝硬化标本中突变型p53蛋白的表达，并探讨后者与肝癌临床病理学特征之间的关系。结果：肝癌突变型p53蛋白表达阳性率为70％（20／29），癌旁组织阳性率为60％（18／29），肝硬化组织阳性率为30％（7／23），有血管侵犯的12例肝癌变变型p53蛋白表达均为阳性，而无血管侵犯的17例中仅8例为阳性（P＜0．05）；突变型p53蛋白阳性组血清甲胎蛋白（AFP）明显高于阴性组（8370±4764ng／ml比98．7±64．3ng／ml，P＜0．05）。提示：P53蛋白表达在肝硬化时即发生了异常；p53蛋白异常可能是AFP基因被激活的原因之一；p53蛋白异常有利于肝癌向门静脉转移。     

慢性淋巴细胞白血病的分子遗传学特点

目的 了解慢性淋巴细胞白血病（CLL）的分子遗传学特性。方法运用间期荧光原位杂交（FISH）技术对60例初发的B细胞CLL（B-CLL）患者进行12号染色体3体（＋12）、del（13q14）和del（17p13）检测。结果60例患者中,41例（68．3％）至少有一种分子遗传学异常,2例（3．3％）具有2种染色体异常。12例（20．0％）有＋12异常,其畸变细胞率在4．0％-34．0％之间;24例（40．0％）有del（13q14）异常,其畸变细胞率在22．0％～93．0％之间,其中3例有2条染色体del（13q14）异常;7例（11．7％）有del（17p13）异常,其畸变细胞率在6．0％～68．0％之间。不同Binet分期中,3种分子遗传学异常差异无统计学意义。结论FISH是一种在分析CLL染色体数目和结构异常方面较为快速、准确和敏感的方法,可为CLL的研究提供较为准确的分子遗传学信息。     

肝细胞癌染色体4q和8p杂合性丢失的研究

目的：检测肝细胞性肝癌患者的4q和8p两个区域上相关的9个微卫星位点的杂合性丢失频率,探讨杂合性丢失与临床病理学特征的关系。方法：选取45例信息性肝癌标本,提取组织DNA,并检测浓度,然后进行PCR扩增,最后将产物通过变性聚丙烯酰胺凝胶电泳分离并观察目的条带有无密度减少或丢失。结果：45例信息性肝癌病例的9个微卫星位点总染合性缺失率为66.7%。其中D8S552的LOH率最高为41.9%,通过χ2检验或Fisher确切概率法进行统计学分析（P〈0.05）,得到该位点在性别（P=0.023）,血清AFP（P=0.004）,有无肝内转移（P=0.023）中均有统计学差异。D4S415为22.5%,D4S3331为22.3%,D8S1810为18.8%,D4S2954为15.4%,D4S3030为15.4%;D8S1725为12.5%,D8S1827为9.8%,D8S1754为6.3%。结论：染色体4q和8p上D4S415、D4S3331、D8S552位点杂合性丢失是肝细胞性肝癌发生发展的重要事件,提示其临近部位可能存在潜在的抑癌基因;为其他肝癌相关抑癌基因的研究提供理论基础。     

前列腺肿瘤高密度allelotype初步分析

目的：探讨原发性前列腺癌及高级别前列腺上皮内肿瘤（PIN）22条常染色体等位基因杂合性缺失及其意义采用显微切割技术提取前列腺癌及高级别前列腺上皮内肿瘤各10例的DNA，然后采用PCR及微卫星多态性技术，对22条常染体上的382个微卫星标志位点进行高密度allelotype分析。结果：10例原发性前列腺癌在染色体1p,1q,2p,2q,6q,8p,8q,10q,11q,13q,16p,17p等部位存在高频杂合性缺失（LOH），前列腺上皮内肿瘤则在染色体2p,2q,6q,8p,10q,1q等区易检测到LOH.结论：前列腺癌患者2q14,8p12-21,10q23-24,13q14等区域存在高频的LOH，位于这些区域的抑癌基因在前列腺癌发生，发展过程中起着重要作用。     

从门静脉癌栓建立肝癌细胞株及对其细胞遗传特征的研究

背景和目的：肝细胞癌（hepatocellular carcinoma,HCC)预后差的原因是术后复发率高和早期过门静脉转移。本研究建立门静脉癌栓肝癌细胞株,并探讨其分子细胞遗传特征。方法：从HCC患者门脉癌栓中获得取肝癌细胞进行细胞培养,对培养成功的细胞（H4M）进行染色体G带染色后分析其核型和对比基因组杂交（comparative genomic hybridization,CGH)结果;用差异PCR检测周期素D1基因。结果：H4M细胞核型为超3倍体,染色体数为71－78,其中有一条标记染色体含有一长的均染区（hsr)。H4M主要的细胞遗传学改变为8p缺失和11q13高拷贝数扩增。周期素D1基因CCND1明显扩增。结论：染色体8p丢失和11q13高拷贝数扩增与H4M细胞的转移特性相关。周期素D1基因CCND1扩增可能是染色体11q13扩增的原因。     

Array-based comparative genomic hybridization reveals recurrent chromosomal aberrations and Jab1 as a potential target for 8q gain in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the major malignancies worldwide. We have previously characterized global gene expression patterns in HCC using microarrays. Here, we report the analysis of genomic DNA copy number among 49 HCC samples using BAC array-based comparative genomic hybridization (CGH). We observed recurrent and characteristic chromosomal aberrations, including frequent DNA copy number gains of 1q, 6p, 8q and 20q, and losses of 4q, 8p, 13q, 16q and 17p. We correlated gene expression with array CGH data, and identified a set of genes whose expression levels correlated with common chromosomal aberrations in HCC. Especially, we noticed that high expression of Jab1 in HCC significantly correlated with DNA copy number gain at 8q. Quantitative microsatellite analysis further confirmed DNA copy number gain at the Jab1 locus. Overexpression of Jab1 in HCC was also validated using real-time RT-PCR, and Jab1 protein levels were studied by immunohistochemistry on tissue microarrays. Functional analysis in HCC cell lines demonstrated that Jab1 may regulate HCC cell proliferation, thereby having a potential role in HCC development. In conclusion, this study shows that array-based CGH provides high resolution mapping of chromosomal aberrations in HCC, and demonstrates the feasibility of correlating array CGH data with gene expression data to identify novel oncogenes and tumor suppressor genes.     

TERT promoter mutations and chromosome 8p loss are characteristic of nonalcoholic fatty liver disease‐related hepatocellular carcinoma

The number of patients with nonalcoholic fatty liver disease (NAFLD)‐related hepatocellular carcinoma (HCC) is increasing. To understand the molecular features of the tumor phenotype, we aimed to clarify the overall landscape of genetic aberrations accumulated in NAFLD‐related HCC. Of 247 HCC patients who underwent hepatectomy during 2010 to 2014 at a single center in Japan, 10 were diagnosed with NAFLD‐HCC based on strict clinical and pathologic criteria. We analyzed the genetic aberrations of 11 NAFLD‐HCC tumor samples from these 10 patients by whole‐exome sequencing, targeted sequencing of the selected genes, and copy number variation studies. Whole‐exome sequencing revealed a mean somatic mutation rate of 1.86 per megabase, and 12 genes were recurrently mutated in NAFLD‐HCCs. Targeted sequencing of the 26 selected genes (12 recurrently mutated genes in whole‐exome sequencing and 14 representative HCC‐associated genes) revealed that TERT promoter mutations occurred in 9 of 11 HCCs (82%), followed by CTNNB1 (45%) and TP53 (36%) mutations. Array‐based copy number variation studies identified recurrent gains at 1q and 8q, and recurrent losses at 1p, 4q, 6q, 8p, 13q, 16p, 17p, and 18q. Notably, chromosome 8p loss occurred in all of the NAFLD‐HCC samples. The current study provided the characteristics of genetic aberrations in NAFLD‐HCC and suggested that TERT promoter mutations and chromosome 8p loss mainly contribute to NAFLD‐related liver carcinogenesis.     

Distinct chromosomal bias of gene expression signatures in the progression of hepatocellular carcinoma

To identify the chromosomal aberrations associated with the progression of liver cancer, we applied expression imbalance map analysis to gene expression data from 31 hepatocellular carcinomas and 19 noncancerous tissues. Expression imbalance map analysis, which detects mRNA expression imbalance correlated with chromosomal regions, showed that expression gains of 1q21-23 (74%), 8q13-21 (48%), 12q23-24 (41%), 17q12-21(48%), 17q25 (25%), and 20q11 (22%) and losses of 4q13 (48%), 8p12-21 (32%), 13q14 (32%), and 17p13 (29%) were significantly associated with hepatocellular carcinoma. Most regions with altered expression identified by expression imbalance map were also identified in previous reports using comparative genomic hybridization. We demonstrated chromosomal copy number gain in 1q21-23 and loss in 17p13 by genomic quantitative PCR, suggesting that gene expression profiles reflect chromosomal alterations. Furthermore, expression imbalance map analysis revealed that more poorly differentiated hepatocellular carcinoma contain more chromosomal alterations, which are accumulated in a stepwise manner in the course of hepatocellular carcinoma progression: expression imbalance of 1q, 8p, 8q, and 17p occur as early events in hepatocarcinogenesis, and 12q, 17q25 and 20q occur as later events. In particular, expression gain of 17q12-21 and loss of 4q were seen to accumulate constantly through the dedifferentiation process. Our data suggest that gene expression profiles are subject to chromosomal bias and that expression imbalance map can correlate gene expression to gene loci with high resolution and sensitivity.     

Chromosomal abnormality in hepatocellular carcinoma by comparative genomic hybridisation in Taiwan

The elucidation of the genetic changes of hepatocellular carcinoma (HCC) is very important for understanding the molecular mechanism of liver carcinogenesis. In order to identify the gains or losses in DNA sequence copy number in HCC, we used comparative genomic hybridisation to study 40 cases (44 tumours) of HCC. Tumour DNA and DNA from non-neoplastic liver tissue were labelled with different fluorochromes and then simultaneously hybridised to normal metaphase spread chromosomes. An image acquisition system was used to quantitate signal intensities contributed by tumour and reference DNA along the entire length of each chromosome. Regions of amplification and deletion were demonstrated as quantitative alterations. Losses were prevalent on chromosome regions 16q (43%), 17p (20%), 13q (20%), 4q (15%) and 8p (15%). Gains frequently occurred on 8q (30%), 1q (20%), 6p (20%) and 17q (18%). Hepatitis B virus carriers had a significantly higher frequency of losses on chromosome 16q. Furthermore, the minimal region of losses was narrowed down to 16q11-q22. This study confirms the presence of previously known chromosomal aberrations in HCC and highlights a new significant correlation between losses on chromosome 16q and hepatitis B virus carriers.     

Can a genetic signature for metastatic head and neck squamous cell carcinoma be characterised by comparative genomic hybridisation?

Survival from head and neck squamous cell carcinoma (HNSCC) has remained static for the last 20 years. The development of lymph node metastasis (LNM) significantly reduces the 5-year survival rate, thus the ability to identify tumours with the potential to metastasise would allow more aggressive treatment regimes to be directed at these patients regardless of negative clinical and radiological findings at the time of presentation. Comparative genomic hybridisation (CGH) can identify chromosomal aberrations that may lead to metastasis. DNA from 23-paired specimens of primary tumour (PT) and LNM were analysed. Nonrandom copy number changes were identified in all paired samples. Similar numbers of aberrations were identified on PT and LNM samples. The most common aberrations were 3q (90%), 8q (65%), 1q (50%), 5p (43%), 2q (41%) and 11q (41%) and deletions 3p (57%), 1p (54%), 4p (48%), 13q (48%), 11q (41%) and 10q (37%). A number of differences were also detected. No aberration was found to be preferentially associated with the LNM, although gains on 6q (48 vs 22%) and 22q (26 vs 9%) were found at higher frequencies. Clonality studies demonstrated that LNM develop from the dominant population of cells in the PT. These results were compared with two similar publications. No combination of chromosomal aberrations, as detected by CGH, was associated with metastatic progression in HNSCC.     

Deletion of 1p32-p36 is the most frequent genetic change and poor prognostic marker in adenoid cystic carcinoma of the salivary glands

PURPOSE: Adenoid cystic carcinoma (ACC) is a relatively uncommon salivary gland malignancy known for its protean phenotypic features and pernicious clinical behavior. Currently, no effective therapy is available for patients with advanced nonresectable, recurrent, and/or metastatic disease. The purpose of this study is to identify prognostic factors other than tumor stage that can be used to predict the outcome of the patients with ACC. EXPERIMENTAL DESIGN: We used comparative genomic hybridization (CGH) to identify copy number aberrations in 53 primary ACCs. Array CGH and fluorescence in situ hybridization analysis was used to validate CGH results on selected cases. We correlated these copy number aberrations with clinicopathologic factors using Pearson's chi2 or by the two-tailed Fisher exact test. The disease-specific survival and disease-free intervals were generated by the Kaplan-Meier product limit method. RESULTS: Chromosomal losses (n = 134) were more frequent than gains (n = 74). The most frequent genetic change was the loss of 1p32-p36 in 44% of the cases followed by 6q23-q27, and 12q12-q14. The most frequently gained chromosomal regions were 8 and 18. Of the chromosomal aberrations, loss of 1p32-p36 was the only abnormality significantly associated with patient's outcome. CONCLUSIONS: This study, for the first time, identifies loss of 1p32-p36 as a significant aberration in ACC. Molecular characterization of 1p32-36 region using the available genomic technologies may lead to the identification of new genes critical to the development of novel therapeutic targets for this disease copy number aberration.     

Array comparative genomic hybridization analysis of colorectal cancer cell lines and primary carcinomas

Array comparative genomic hybridization, with a genome-wide resolution of approximately 1 Mb, has been used to investigate copy number changes in 48 colorectal cancer (CRC) cell lines and 37 primary CRCs. The samples were divided for analysis according to the type of genomic instability that they exhibit, microsatellite instability (MSI) or chromosomal instability (CIN). Consistent copy number changes were identified, including gain of chromosomes 20, 13, and 8q and smaller regions of amplification such as chromosome 17q11.2-q12. Loss of chromosome 18q was a recurrent finding along with deletion of discrete regions such as chromosome 4q34-q35. The overall pattern of copy number change was strikingly similar between cell lines and primary cancers with a few obvious exceptions such as loss of chromosome 6 and gain of chromosomes 15 and 12p in the former. A greater number of aberrations were detected in CIN+ than MSI+ samples as well as differences in the type and extent of change reported. For example, loss of chromosome 8p was a common event in CIN+ cell lines and cancers but was often found to be gained in MSI+ cancers. In addition, the target of amplification on chromosome 8q appeared to differ, with 8q24.21 amplified frequently in CIN+ samples but 8q24.3 amplification a common finding in MSI+ samples. A number of genes of interest are located within the frequently aberrated regions, which are likely to be of importance in the development and progression of CRC.     

Genome-wide-array-based comparative genomic hybridization reveals genetic homogeneity and frequent copy number increases encompassing CCNE1 in fallopian tube carcinoma

Fallopian tube carcinoma (FTC) is a rare, poorly studied and aggressive cancer, associated with poor survival. Since tumorigenesis is related to the acquisition of genetic changes, we used genome-wide array comparative genomic hybridization to analyse copy number aberrations occurring in FTC in order to obtain a better understanding of FTC carcinogenesis and to identify prognostic events and targets for therapy. We used arrays of 2464 genomic clones, providing approximately 1.4 Mb resolution across the genome to map genomic DNA copy number aberrations quantitatively from 14 FTC onto the human genome sequence. All tumors showed a high frequency of copy number aberrations with recurrent gains on 3q, 6p, 7q, 8q, 12p, 17q, 19 and 20q, and losses involving chromosomes 4, 5q, 8p, 16q, 17p, 18q and X. Recurrent regions of amplification included 1p34, 8p11-q11, 8q24, 12p, 17p13, 17q12-q21, 19p13, 19q12-q13 and 19q13. Candidate, known oncogenes mapping to these amplicons included CMYC (8q24), CCNE1 (19q12-q21) and AKT2 (19q13), whereas PIK3CA and KRAS, previously suggested to be candidate driver genes for amplification, mapped outside copy number maxima on 3q and 12p, respectively. The FTC were remarkably homogeneous, with some recurrent aberrations occurring in more than 70% of samples, which suggests a stereotyped pattern of tumor evolution.     

CD147和CK19在肝细胞癌中的表达及临床意义

目的 探讨细胞外基质蛋白诱导因子（CD147）和细胞角蛋白19（CK19）在肝细胞癌（HCC）中的表达及临床意义。方法 采用组织芯片技术和免疫组织化学法检测CD147和CK19在272例HCC组织和81例癌旁组织中的表达情况。结果 CD147在HCC中的阳性表达率为73.53％（200／272）,在癌旁组织中的阳性表达率为13.58％（11／81）,差异有统计学意义（P＜0.05）。CK19在HCC组织中的阳性表达率为14.34％（39／272）,CK19在癌旁组织中无表达。在HCC中,CD147的表达与组织学分级、临床分期、术后无瘤生存时间、肿瘤直径、脉管或门静脉癌栓相关,与性别、年龄、肝硬化、AFP水平、HBV感染、淋巴结转移、病灶数目、肝被膜浸润及卫星灶无关（P＞0.05）;CK19在HCC中的表达与术后无瘤生存时间、组织学分级、肿瘤直径、肝硬化、卫星灶、淋巴结转移及临床分期有关,与性别、年龄、病灶数目、肝被膜浸润、AFP水平、HBV感染及脉管或门静脉癌栓无关（P＞0.05）。在HCC中,CD147 阳性表达者与阴性表达者的中位复发时间分别为13个月和48个月（P＜0.05）,中位生存时间分别为24个月和60个月（P＜0.05）;CK19阳性表达者与阴性表达者的中位复发时间分别为7个月和31个月（P＜0.05）,中位生存时间分别为13个月和42个月（P＜0.05）。 CD147的表达和CK19的表达无明显相关性（r=0.061,P=0.317）。结论 HCC中CD147和CK19的表达与预后密切相关,且两者均可作为HCC预后不良的判断指标。     

Molecular karyotyping of human hepatocellular carcinoma using single-nucleotide polymorphism arrays

Genomic amplification of oncogenes and inactivation of suppressor genes are critical in the pathogenesis of human cancer. To identify chromosomal alterations associated with hepatocarcinogenesis, we performed allelic gene dosage analysis on 36 hepatocellular carcinomas (HCCs). Data from high-density single-nucleotide polymorphism arrays were analysed using the Genome Imbalance Map (GIM) algorithm, which simultaneously detects DNA copy number alterations and loss of heterozygosity (LOH) events. Genome Imbalance Map analysis identified allelic imbalance regions, including uniparental disomy, and predicted the coexistence of a heterozygous population of cancer cells. We observed that gains of 1q, 5p, 5q, 6p, 7q, 8q, 17q and 20q, and LOH of 1p, 4q, 6q, 8p, 10q, 13q, 16p, 16q and 17p were significantly associated with HCC. On 6q24-25, which contains imprinting gene clusters, we observed reduced levels of PLAGL1 expression owing to loss of the unmethylated allele. Finally, we integrated the copy number data with gene expression intensity, and found that genome dosage is correlated with alteration in gene expression. These observations indicated that high-resolution GIM analysis can accurately determine the localizations of genomic regions with allelic imbalance, and when integrated with epigenetic information, a mechanistic basis for inactivation of a tumor suppressor gene in HCC was elucidated.