乳腺癌中bcl-2表达与p53和雌激素受体关系的研究

目的　研究乳腺癌中bcl- 2基因表达与p5 3基因和雌激素受体的关系以及对乳腺癌生物学行为影响。方法　应用S -P免疫组织化学方法对 4 5例乳腺癌石蜡包埋组织进行检测。结果在人的乳腺癌中bcl- 2、p5 3蛋白和雌激素受体阳性表达率分别为 5 5 5 %、5 1 1%和 4 6 7%。bcl- 2阳性表达与肿瘤大小、淋巴结是否发生转移无关 (P >0 0 5 ) ;与p5 3阳性表达呈负相关 (P <0 0 5 ) ;与乳腺癌恶性程度和雌激素受体阳性表达呈正相关 (P <0 0 5 )。结论　bcl- 2基因、p5 3基因和雌激素共同参与了乳腺癌的发生和发展 ;bcl- 2阳性表达常会减弱p5 3的表达 ;为综合评价bcl- 2基因在乳腺生物学行为中的作用提供了一些线索。     

免疫组化和原位杂交技术联合检测sFLK-1片段基因转染后的表达

血管内皮细胞生长因子 (vascularendothelialcellgrowthfactor,VEGF)与其受体Flt 1、FLK 1/KDR的相互作用是调控肿瘤血管形成的关键 ,阻断VEGF与相应受体的结合成为目前抗肿瘤血管形成生物学治疗的热点。本文将转染至肿瘤细胞系S180和B16的外源基因sFLK 1片段接种到纯系小鼠BABL/C和C5 7的皮下 ,观察转染后基因分别在蛋白质和mRNA水平上的表达特点。1　材料与方法1.1　材料及试剂　细胞株 :B16 sFLK 1fragment、S180 sFLK 1fragment、B16 空载体、S180 空载体为本实验获得。各相应对照亲代细胞株为本研究室保存。纯系B…     

宫颈鳞状细胞癌组织中TGF-β_1、p53基因的原位杂交检测

宫颈癌是最常见的女性生殖道恶性肿瘤，死亡率居妇科肿瘤第２位，其发生是多因素、多步骤的复杂过程〔１〕。通过研究宫颈鳞状细胞癌组织中ＴＧＦ－β１、ｐ５３基因的变化，进一步揭示宫颈癌发生的分子机制。１材料和方法１．１标本２２例宫颈鳞状细胞癌均为本院妇产科手...     

同源异型盒基因HOXB7在乳腺癌组织中的表达及其临床意义

<正>目的:乳腺癌是女性最常见的恶性肿瘤之一,并且有发病年龄提早的趋向,因此研究促进乳腺癌发生的因素显得尤为重要。HOXB7是同源异型盒基因家族中新近发现的一种转录因子,它在生理状态下的表达对维持正常细胞的生长和分化有非常重要的作用,然而其异常表达时可能促进肿瘤形成。本实验探讨HOXB7在乳腺癌组织中的表达及其临床意义。方法::收集正常乳腺组织20例、乳腺纤维瘤组织20例、癌组织60例,应用实时荧光定量逆转录聚合酶链反应(Real-time RT-PCR)检测三种乳腺组织中HOXB7 mR-NA的表达;应用Western Blotting法检测三种乳腺组织中HOXB7蛋白的表达水平     

原位杂交检测乳腺癌雌激素受体mRNA的表达

目的：检测乳腺癌组织中雌激素受体ｍＲＮＡ的表达状况。方法：采用地高辛配基标记探针的非同位素原位杂交技术检测了１５列乳腺癌组织冰冻切片。结果：蓝紫色的阳性染色颗粒位于癌细胞的胞浆及胞核，背景清晰。并且与葡萄糖包裹活性碳及免疫组化法检测雌激素受体的阳阴性符合率分别为９３．３％和１００％。结论：该方法是一特异、灵敏、简单易行的研究雌激素受体ｍＲＮＡ表达水平的有效手段。     

人乳腺癌HPV感染的超微结构和DNA分子原位杂交研究

目的：对乳腺癌的HPV感染进行形态学定位研究。方法：对46例乳腺癌切除标本及7例良性病变标本进行透射电子显微镜以及HPV DNA分子原位杂交检测。结果：透射电镜下，14例（30．40％）乳腺癌细胞核内有HPV样病毒颗粒，颗粒直径30－50nm，或弥漫散布，或聚集成团呈假结晶状排列。颗粒圆形或略不规则，含有HPV样颗粒的癌细胞核异型性较明显。7例良性病变细胞核内未见到HPV样颗粒。HPV DNA分子原位杂交显示，21例（45．65％）乳腺癌细胞核内HPV31／33 DNA阳性，1例软肉瘤样化生性癌细胞核同时有HPV31／33及HPV16／18阳性；1例纤维腺癌（14．29％）也显示HPV31／33 DNA阳性（X^2=2．7431，P＜0．05）。电镜下85％病毒样颗粒阳性病例中可检测到HPV DNA存在，两种检测结果符合率达73．91％。结论：乳腺肿瘤内存在未组装的、裸型病毒样颗粒。该研究对阐明乳腺癌的发病原因以及对乳腺癌的预防和治疗均有重要意义。     

Chromogenic and fluorescent in situ hybridization in breast cancer

Fluorescent (FISH) and chromogenic (CISH) in situ hybridization have recently become part of the diagnostic armamentarium of breast pathologists. HER2 gene testing by FISH and/or CISH has become an integral part of the diagnostic workup for patients with breast cancer. In this era of high throughput technologies, these techniques have proven instrumental for the validation of results from microarray-based comparative genomic hybridization and for the identification of novel oncogenes and tumor suppressor genes. Furthermore, FISH and CISH applied to tissue microarrays have expedited the characterization of genomic changes associated with specific breast cancer molecular subtypes and the identification of novel prognostic and predictive markers. In this review, we provide in this review a critical assessment of CISH and FISH and the impact of the analysis of amplification of specific oncogenes (eg, HER2, EGFR, MYC, CCND1, and FGFR1) and deletion of tumor suppressor genes (eg, BRCA1 and BRCA2) on our understanding of breast cancer.     

Detection of numerical alterations of chromosome 1 in cytopathological specimens of breast tumors by chromogen in situ hybridization

To investigate the effectiveness of chromogen in situ hybridization (CISH) in the diagnosis of breast tumors, numerical alterations of chromosome 1 were examined by CISH and fluorescence in situ hybridization (FISH) methods, and the presence of der(16)t(1;16) was also examined by FISH in imprinted cytology specimens from resected tissues of 14 carcinomas and five non-malignant lesions. The modal signal counts of chromosome 1 were compared between the specimens processed by CISH and FISH for each case. Aneusomies of the long arm of chromosome 1 were detected in 10 (71%) carcinomas as the major clones by both methods. In addition, one atypical papilloma demonstrated tetrasomy of 1q12 as a major clone by CISH, but such a clone was at first overlooked by FISH. Four other benign lesions showed disomic 1q12 signals as a major clone by both CISH and FISH. As additional information from FISH, eight cancers showed structural or numerical alterations of chromosome 16, and four showed der(16)t(1;16). In total, 10 carcinomas showed chromosome 16 alterations, and all of these overlapped with the carcinomas with 1q12 aneusomies. The CISH method provided almost the same results as the FISH method, and both methods were considered applicable in supportive diagnosis of cytological specimens of breast tumors. In addition, the CISH method was superior in the detection of numerical alterations in carcinoma cells by referring to the morphology of cells and in the detection of significant clones which might be missed under dark-field microscopy.     

Ⅰ型细胞间粘附分子在肝细胞癌中的表达及其意义

目的探讨肝癌细胞产生的Ⅰ型细胞间粘附分子１（ＩＣＡＭ－１）在肝细胞癌侵袭和转移过程中的作用。方法用原位分子杂交技术检测了３４例原发性肝癌及癌旁组织中ＩＣＡＭ－１ｍＲＮＡ的表达。结果２１例侵袭性生长的肝癌中１９例ＩＣＡＭ－１ｍＲＮＡ表达阳性,明显高于癌旁;１１例伴门静脉癌栓或肝内转移灶形成的肝癌,其癌组织均表达强阳性,癌栓及转移灶表达阳性;而１３例包膜完整的肝癌仅有４例表达阳性。结论肝癌细胞ＩＣＡＭ－１ｍＲＮＡ的表达与肝癌细胞的侵袭及转移相关。     

乳腺癌miRNAs表达谱的检测

目的检测人乳腺癌石蜡组织及人乳腺癌细胞株中miRNAs的表达情况,初步筛选人乳腺癌miRNAs表达谱。方法(1)应用组织微阵列平台,采用原位分子杂交技术对91例人乳腺癌和26例癌旁乳腺组织标本42种miRNA进行检测;(2)培养人乳腺癌细胞株MDA-MB-231和正常乳腺上皮细胞株HBL-100,分别抽提细胞总RNA,分离小分子RNA,荧光标记后与miRNAs微阵列杂交,通过芯片扫描和数据分析,获得人乳腺癌miRNAs表达谱。结果(1)与癌旁乳腺组织相比,14种miR-NAs在乳腺癌组织中的表达发生了显著变化(P0.05),其中9种表达上调,5种表达下调。(2)利用miRNAs微阵列,筛选获得71种与人乳腺癌相关miRNAs,与正常乳腺上皮细胞相比,34种miRNAs表达上调,37种表达下调。结论筛选出与乳腺癌相关的差异表达miRNAs,为进一步研究其在乳腺癌中的作用奠定基础。     

生物素标记探针原位杂交法检测细胞中c-myc基因的表达

本文采用生物素标记的c-mye基因作探针，通过细胞原位分子杂交技术，对人舌癌、膀胱癌和正常细胞中c-myc基因的转录量进行了定性和半定量分析。结果表明与正常细胞相比，舌癌和膀胱癌细胞中c-myc基因异常高度表达。     

肺癌组织p16和Rb基因mRNA表达的双重原位杂交观察

目的：研究Ｐ１６和Ｒｂ基因在ｍＲＮＡ表达及其与肺癌的关系。方法：制备ｐ１６和Ｒｂ基因ｃＤＮＡ探针,采用双重原位杂交的方法,地８９例肺癌和正常肺组织进行了Ｐ１６ 一Ｒｂ基因ｍＲＮＡ表达的定位研究。结果：小细胞肺癌ｐ１６ｍＲＮＡ阳性表达率高达８２．４％,而非小细胞肺癌阳性率为４５．８％,两者差异有显著性。非小细胞肺癌ＲｂｍＲＮＡ阳性表达率为８１．９％,而小细胞肺癌一率仅５．９％,两者差异有高度显著性,     

Array-based comparative genomic hybridization of ductal carcinoma in situ and synchronous invasive lobular cancer

It has been increasingly recognized that ductal carcinoma in situ (DCIS), lobular carcinoma in situ (LCIS) and invasive cancer of the breast are often closely associated with one another. However, the genomic relationship between these histologically distinct entities has not been well characterized. Refinements in high-resolution comparative genomic hybridization (CGH) techniques allow for a detailed comparison of genomic alterations in synchronously occurring tumors. The following case illustrates how array CGH may be used to better understand whether synchronous neoplasms share a common origin.     

Evaluation of HER2 gene amplification in invasive breast cancer using a dual‐color chromogenic in situ hybridization (dual CISH)

Fluorescence in situ hybridization (FISH) assay is considered the ‘gold standard’ for evaluation of HER2/neu (HER2) gene status, however, it is difficult to recognize morphologic features of tumors using fluorescence microscopy. Thus, chromogenic in situ hybridization (CISH) has been proposed as an alternative method to evaluate HER2 gene amplification. Here, we examined the dual color CISH (dual CISH) method which provides information regarding the copy number of the HER2 gene and chromosome 17 centromere from a single slide. We examined 40 cases of invasive ductal carcinomas of the breast that were resected surgically. HER2 gene status was assessed with FISH (Abbott) and dual CISH (Dako). HER2 gene amplification status was classified according to the guidelines of the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP). Comparison of the cut‐off values for HER2/chromosome 17 centromere copy number ratio obtained by dual CISH and FISH showed that there was almost perfect agreement between two methods (Kappa coefficient 0.96). The results of the two commercial products were almost consistent for evaluation of HER2 gene counts on the sections. The current study proved that dual CISH is comparable with FISH for evaluating HER2 gene status.