肝内胆管癌多基因变异表型分析

目的：探讨参与肝内胆管癌发生的多基因变异谱系,为了肝内胆管癌的发生机理提供基因水平上的依据。方法：采用显微切割术从石蜡组织切片中提取DNA、聚合酶链式反应（PCR）扩增基础上DNA测序的方法,对22例手术切除的肝内胆管癌标本进行了6种肿瘤抑制基因（APC、MCC、DCC、OGG1、p53和RB1）的杂合性缺失（LOH）和Ki-ras-2癌基因点突变的发生率检测。结果7种肿瘤基因的变异率为APC（68．8％）、DCC（46．2％）、OGG1（41．7%）、p53(37.5%)、Ki-ras-2(27．3％）、RB1（22．2％）和MCC（14．3％）。结论多基因变异的协同作用在肝内胆管癌的多阶段发展发生过程中起着重要作用,其中尤以APC、DCC、OGG1、p53和Ki-ras-2构成了肝内胆管癌相关的基本基因谱系。     

毛细管电泳法检测肝细胞癌微卫星基因表型的影响因素

目的：分析影响肝细胞癌微卫星—自动毛细管电泳检测结果的技术因素。方法：应用MegaBACE500毛细管电泳测序仪对一组高度多态性微卫星PCR扩增产物进行电泳，应用Genetic Profiler软件进行基因表型分型，比较PCR扩增产物在不同处理条件下对基因分型的影响。结果：PCR反应体系残留混合物(dNTP、引物和盐离子的浓度等)对基因分型的结果有明显的影响，当PCR体系中益离子和DNA的浓度比＞10000：1时可使等位基因片段分析系统评分出现偏差而导致结论错误，基因组DNA—PCR样品浓度在50ng/μl时较为合适。结论：毛细管电泳测序体系中PCR反应体系残留混合物的浓度是影响基因分型结果的一个重要因素。     

肝细胞癌和癌旁肝组织中PTEN蛋白表达及其临床意义

目的：探讨肝细胞癌（HCC）和癌旁肝组织中PTEN蛋白表达及其临床意义。方法：利用SP免疫组织化学技术检测PTEN蛋白在HCC及其癌旁肝组织的表达。结果：PTEN蛋白在HCC中的阳性率和阳性强度均低于癌旁肝组织（P＜0．01）,HCC中PTEN蛋白的表达强度与患者的性别和肿瘤的大小无关（P＞0．05）,但与肿瘤细胞的分化程度明显相关（P＜0．01）,分化愈差,表达愈强。结果：提示检测PTEN蛋白可能对判断HCC的预后具有一定意义。     

肝细胞癌及癌旁肝组织中PTEN表达与MAPK磷酸化的相关性

目的：研究肝细胞癌（HCC）及癌旁肝组织中PTEN表达与丝裂素激活的蛋白激酶（mitogen-activated protein kinase,MAPK)磷酸化的相关性。方法：利用SP免疫组织化学检测75例HCC及其癌旁肝组织中PTEN蛋白和p42/44^MAPK的表达。结果：①HCC中PTEN蛋白的阳性率（62．7％）和阳性强度明显低于癌旁肝组织（89．3％）（P＜0．01），且阳性信号强度与HCC分化程度相关（P＜0．01），即HCC分化愈差，PTEN蛋白表达愈弱。②HCC中p42/44^MAPK的阳性率（85．3％）和表达强度明显高于癌旁肝组织（34．7％）（P＜0．01），但阳性信号强度与癌细胞分化程度无关（P＞0．05）。③相关性分析表明，HCC及癌旁肝组织中PTEN蛋白表达强度和MAPK磷酸化程度呈明显负相关（P＜0．01）。结论：提示在HCC发生中，可能是由于PTEN蛋白表达降低或缺失，不能有效抑制由致癌因子异常激活的Ras/Raf/MAPK信号转导通路，从而使肝细胞异常增殖和发生恶性转化。     

用荧光差异显示法分离新的肝细胞癌相关基因

目的：寻找新的肝细胞癌（ｈｅｐａｔｏｃｅｌｌｕｌａｒ ｃａｒｃｉｎｏｍａ,ＨＣＣ）相关基因。方法：应用荧光－差异显示（ｆｌｕｏｒｅｓｃｅｎｔ ｄｉｆｆｅｒｅｎｔｉａｌ ｄｉｓｐｌａｙ）技术分析人ＨＣＣ、配对非癌肝、胎肝、正常肝等组织之间的基因差异表达,差异表达ｃＤＮＡ片段,经测序后,再用ＲＮＡ狭缝杂交对部分差异片段在上述４种组织中的表达进行验证。结果：共得到差异表达ｃＤＮＡ片段１１０条。经狭缝杂交     

肝细胞癌患者免疫治疗前后表型分析

为探讨McAb激活T细胞,加强APC的递呈抗原的能力,采用CD28／CD80(受体与配体)McAb共激活的健康人的外周血淋巴细胞输入肿瘤患者,用FACS检测健康人、肿瘤患者治疗前后的PBLs的表型;结果显示在治疗前,肝癌患者PBLs有CD4偏移现象,而治疗后CD4偏移消失,CD3降低,且杀伤细胞(CD8^ )增多。结论：通过共激活PBLs输入疗法,激活了T细胞某些被抑制了的功能,使不同的T细胞向相应细胞转化,增强杀伤肿瘤细胞的作用。     

肝门胆管内肝细胞癌癌栓的磁共振诊断

鉴于国内尚未见磁共振 (ＭＲＩ)诊断肝胆总管内肝细胞癌 (ＨＣＣ)癌栓 (以下简称ＨＣＣ癌栓 )的专门报道 ,特将我院近来所见 2例作一报告 ,以期提高ＭＲＩ对本病的诊断水平。1　临床资料　　例 1.4 8岁 ,男 ,因进行性无痛性黄疸 2 0ｄ被当地诊为“梗阻性黄疸”治疗无效转入我院。入院查血清总胆红素 (Ｔ -ＢＩＬ) 2 71 5 μｍｏｌ/Ｌ(正常 0～ 35 μｍｏｌ/Ｌ) ,结合胆红素 (Ｄ -ＢＩＬ) 16 7 7μｍｏｌ/Ｌ(正常 0～ 2 0 5 μｍｏｌ/Ｌ) ,ＭＲＩ检查发现第一肝门区偏右侧有一 2 5ｃｍ× 2 8ｃｍ大小、呈Ｔ1Ｗ1低信号、Ｔ2 Ｗ1略高信号、…     

肝细胞癌相关差异表达基因和药物预测的生物信息学分析

目的 通过生物信息学方法筛选肝细胞癌(HCC)相关差异表达基因(DEGs)及潜在治疗药物.方法 从GEO数据库中选取GSE45436、GSE84402、GSE62232和GSE101685,使用R软件分析得到DEGs.随后对DEGs进行GO和KEGG分析,并构建PPI网络、筛选核心基因.最后进行生存分析和筛选潜在治疗药...     

肝细胞癌微卫星变异的研究

目的:探讨肝细胞癌(HCC)微卫星变异的特点及其与临床病理的相关性。方法:采用MegaBACE 500型自动化DNA分析系统,对56例HCC中10个微卫星的杂合性缺失(loss of heterozygosity,LOH)、微卫星不稳定性(microsatillite instability,MSI)和等位基因失衡(allelic imbalance,AI)3种变异特征进行检测。结果:56例HCC中,76.8％(43/56)出现LOH,其中以RIZ[76.7％(23/30)]和D4S426[61.0％(25/41)]最高;EdmondsonⅢ、Ⅳ级肿瘤D4S426的LOH明显高于Edmondson Ⅰ、Ⅱ级[76.7％(23/30)vs 18.2％(2/11),P<0.01];血清HBsAg阳性患者中D8S277的LOH发生率明显高于阴性患者[72.7％(16/22)vs12.5％(2/16),P<0.01]。MSI的发生率为32.1％(18/56),其中以BAT-26[24.3％(9/37)]和D13S170[21.9％(7/32)]最高,HBsAg阳性者MSI发生频率显著高于HBsAg阴性者(P<0.01)。AI的发生率为35.7％(20/56)。结论:HCC存在广泛的微卫星变异,其中以代表肿瘤抑制基因路径的LOH方式在HCC的发生和发展过程中起重要作用,代表错配修复基因路径的MSI的作用次之。     

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…     

Two closely neighboring hepatocellular carcinomas mimicking intrahepatic metastasis are confirmed as double primary tumors by loss of heterozygosity analysis of microsatellites

Two spatially-close hepatocellular carcinomas (HCCs) in close proximity to each other could be either one single primary tumor with an intrahepatic metastasis or two independent primary tumors. The authors present a case of a 48-year-old man who underwent a right hepatectomy for two nodules of hepatocellular carcinoma distributed in a sprouting-like pattern mimicking suggestive of satellite nodules, measuring 13.2×9 cm and 4.7×4.3 cm, respectively. Microscopically, the larger HCC showed was (well/moderately/ poorly differentiated) with thick trabecular morphology with grade Ⅲ appearance, and the smaller tumor presented wasa (well/moderately/ poorly differentiated) with a thin trabecular patternn with grade Ⅱhistology. Clonality studies for both nodules by microsatellite-based analysis of loss of heterozygosity (LOH) showed distinctive LOH patterns between them that clearly demonstrated that the two nodules were derived from two separate primary tumor clones. THere, the authors recommend the routine that clonality determination of clonality for multiple satellite-like tumors could providebecause it may provide useful information that will help tofor selecting individual therapeutic options and predicting postoperative outcomes.     

A genome-wide allelotype study of primary and corresponding recurrent glioblastoma multiforme in one patient

Glioblastoma multiforme (GBM) is the most common type of primary malignant brain tumor. Although comprehensive therapeutic measures are available, recurrence is very frequent and the prognosis of GBM remains dismal. To date, little is known about the molecular pathogenesis associated with GBM recurrence. According to Knudson ' s two-hit hypothesis of tumor suppressor gene (TSG) inactivation,1 deletion of a chromosomal region, as revealed by loss of heterozygosity (LOH), is often indicative of the presence of a potential TSG. Allelotype studies involving a comprehensive LOH analysis of the whole genome can provide more detailed and thorough information for detecting genetic anomalies than traditional LOH analysis. The present study is designed to conduct a genome-wide allelotype analysis of one patient ' s primary and corresponding recurrent GBM tumors in an effort to reveal molecular genetic alterations associated with the recurrence of this malignancy.     

A genome-wide allelotype study of primary and corresponding recurrent glioblastoma multiforme in one patient

Glioblastoma multiforme (GBM) is the most common type of primary malignant brain tumor. Although comprehensive therapeutic measures are available, recurrence is very frequent and the prognosis of GBM remains dismal. To date, little is known about the molecular pathogenesis associated with GBM recurrence.     

胶质母细胞瘤染色体13q上肿瘤抑制基因存在的可能性

目的寻找胶质母细胞瘤(glioblastoma,GBM)13号染色体上可能存在肿瘤抑制基因的杂合性丢失 (LOH)区域,为发现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,分析了20例GBM13号染色体上14个微卫星多态性标记的 LOH。结果在60％GBM的13号染色体上观察到LOH,在45.8％可提供信息位点存在LOH。其中13q的 LOH率明显高于13p,13q和13p的LOH率分别为60％、27％。在13q上的下列位点上检测到较高的LOH率 (＞50％)13q14.1-14.3上D13S153,13q12-14.2上的D13S217-D13S263,13q21.2-32上的D13S156-D13S265。 结论13号染色体可能在GBM的分子发病机制中发挥着重要作用,在13q14.1-14.3上的D13S153位点、13q 12-14.2上的D13S217-D13S263和13q21.2-32上的D13S156-D13S265间区域可能存在多个与GBM相关的肿瘤 抑制基因,可能包括RB1以及不同于RB1的其他肿瘤抑制基因。     

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     

Genome-wide allelotype study of primary glioblastoma multiforme

Objective To investigate the molecular genetic pathogenesis of primary glioblastoma multiforme (GBM) and identify which chromosomes or chromosomal regions of the entire genome may harbor tumor suppressor genes (TSGs) associated with GBM.Methods A high-resolution allelotype study of 21 cases of primary GBM was performed by PCR-based loss of heterozygosity （LOH）analysis. Three hundred and eighty-two fluorescent dye-labeled microsatellite markers covering all 22 autosomes were applied. The mean genetic distance between two flanking markers was about 10 cM.Results LOH was observed on all 39 nonacrocentric autosomal arms examined in this study. The LOH frequencies of 10q, 10p, 9p, 17p and 13q were the highest (&gt;50%). Furthermore, high LOH frequencies were detected in the regions containing known TSGs including PTEN, DMBT1, p16, p15, p53 and RB; the LOH frequencies on 14q, 3q, 22q, 11p, 9q, 19q were also high (&gt;40.5%). Our study observed the following commonly deleted regions: 9p22-23, 10p12.2-14, 10q21.3, 13q12.1-14.1, 13q14.3-31, 17p11.2-12, 17p13, 3q25.2-26.2, 11p12-13, 14q13-31, 14q32.1, 14q11.1-13, 22q13.3, 4q35, 4q31.1-31.2, 6q27 and 6q21-23.3. Conclusions The molecular pathogenesis of GBM is very complicated and associated with a variety of genetic abnormalities on many chromosomal arms. The most closely related chromosomal arms to the pathogenesis of GBM are 10q, 10p, 9p, 17p and 13q. Besides the well-known TSGs including PTEN, DMBT1, p16, p15, p53 and RB, multiple unknown TSGs associated with GBM may be present on the commonly deleted regions detected in the present study.     

Two closely neighboring hepatocellular carcinomas mimicking intrahepatic metastasis are confirmed as double primary tumors by the loss of heterozygosity analysis of microsatellites

Hepatocellular carcinoma (HCC) is a common malignancy around the world.Among all HCCs,multifocal HCC accounts for 12％-19％.1 The origin of multifocal HCC can either be the intrahepatic metastasis of a single tumor clone,or independent primaries.2 It is crucial to distinguish intrahepatic metastatic tumor from multicentric HCC because this distinction has significant impact on tumor staging,treatment plan,and clinical outcome.Routine histological examination alone cannot determine the clonal origin of multifocal HCC and usually results in their arbitrary classification as satellite nodules.The only way to reliably determine clonality among multiple lesions is by molecular genetic analysis.We report a patient who underwent surgical resection for two neighboring hepatocellular carcinomas that were from independent primaries.These separate nodules mimicked a solitary primary with intrahepatic metastasis.Confirmation of separate clones was established by the loss of heterozygosity (LOH) of microsatellites.