用荧光原位杂交方法检测肺癌印片标本中性染色体数目的异常

目的 研究肺癌细胞中性染色体拷贝数的异常。方法 取新鲜的肺癌手术标本作印片,利用双色荧光原位杂交（ＦＩＳＨ）技术对１６例肺癌印片标本（其中男性病例１２例,女性病例４例）中Ｘ染色体和Ｙ染色体数目进行检测。结果 发现１２／１６（７５％）的标本中有Ｘ染色体数目增多,其中女性患者３例,男性患者９例;在１２例男性患者中,有１例发生Ｙ染色体数目增多,有５例（４１．７％）发生Ｙ染色体数目减少。总计出现性染色体数     

312例反复流产夫妇的染色体研究

本文对３１２对反复自然流产的夫妇进行了遗传咨询及Ｇ显带染色体研究，发现３２例染色体异常（５．１３％），其中常染色体平衡易位７例（１．１２％），罗伯逊易位１１例（１．７６％），易位核型涉及１、２、４、６、７、９、１１、１３、１４、１５、１７、１８、２１、２２各号染色体，其它异常１４例。     

应用FISH技术检测肺腺癌,鳞癌细胞部分染色体数量的改变

目的研究肺癌细胞７号，１７号及Ｙ染色体的数目改变及其意义。方法应用荧光原位杂交技术（ＦＩＳＨ）技术，对１７例腺癌和鳞癌的细胞印片进行检测。结果７号染色体：三体细胞占细胞总数的３１．７３％。１７号染色体：二体细胞占细胞总数的４３％。Ｙ染色体：单体细胞占细胞总数的５８．０４％。结论在肺腺癌和肺鳞癌中，７号染色体数目均有明显增加，并以三体所占比例最高。肺腺癌与肺鳞癌７号染色体数目分布差异明显。     

314例早期自然流产绒毛的染色体分析

本文对３００名早期自然流产患者的３１４例（１４名两次流产）绒毛标本的染色体进行分析,检出异常核型１３０例,检出率４１．４０％,涉及的异常染色体有１、２、３、４、６、７、８、９、１０、１１、１３、１４、１５、１６、１８、２０、２１、２２、Ｘ．Ｙ。     

苏州地区132例染色体异常分析

苏州市妇幼保健院于１９８５１９９８ 年３ 月对１２６９ 例外周血淋巴细胞作染色体检查,查出异常核型１３２ 例,异常检出率１０．４０％ 。其中常染色体异常５３ 例占４０．１５％ 、性染色体异常７９ 例占５９ ．８５％ ;单纯数目异常８５ 例占６４．３９％ 、结构异常３０ 例占２２ ．７３ ％ 、嵌合型６ 例占４．５５％ 、性别倒错１１ 例占８．３３％ 。按疾病分类：克氏综合征５１ 例（ 标准型５１ 例） ;唐氏综合征２４ 例（标准型２１ 例,嵌合型２ 例,易位型１ 例）;吐纳氏综合征１４ 例（ 标准型７ 例,嵌合型３ 例,Ｘ 畸变４ 例;染色体易位１４ 例（ 平衡易位７ 例,罗伯逊易位６ 例）;４６,ＸＹ女性７ 例,４６,ＸＸ 男性４ 例;１８ 三体３ 例;臂间倒位３ 例;４６ ,ＸＸ（Ｙ） ,１０ｐ ＋ ３例;４７ ,ＸＸＸ２ 例;４６,ＸＹ,２２Ｓ＋ ２ 例;４７,ＸＸ（Ｙ）＋ ｍａｒ２ 例;４６,ＸＹ,１４ｐ ＋１ 例;４６ ,ＸＹｑ－１ 例;４６ ,ＸＸｄｅｌ（４）１ 例。     

2例宫颈癌和1例皮肤癌的衍生染色体

２例宫颈癌和１例皮肤癌的衍生染色体ｄｅｒ（１７；２２）（ｑ１０；ｑｌｏ）［英ｖＮｉｅｌｓＢＡ…／／ＣａｎｃｅｒＧｅｎｅｔＣｙｔｏｇｅｎｅｔ．－１９９４’７４．－１５３。１５５以前，作者已报道显例宫颈癌见到１３例染色体重排（占４２％），导致１７Ｐ丢失，...     

7型腺病毒疫苗株基因克隆及其右侧末端Sma I H片段 …

目的 获得７型腺病毒（Ａｄ７）载体构建的基本元件，为构建Ａｄ７载体及阐明其基因组一级结构打下基础。方法 用改良Ｈｉｒｔｓ法提取Ａｄ７ＤＮＡ，酶切后克隆于质粒载体，克隆了Ｘｈｏ Ｉ Ａ＋Ｄ片段（１７．０￣７６．５ｍｕ）、Ｄ片段（１７．０－２２．９ｍｕ）、Ｅ片段（１０．８－１４．１ｍｕ）、Ｆ＋Ｇ片段（１４．１￣１７．０ｍｕ）、Ｇ片段（１５．８￣１７．０），用ＥｃｏＲ Ｉ＋Ｘｈｏ Ｉ双酶切，克隆了７６．     

应用引物原位标记技术快速检测X和18号染色体

目的为发展快速检测人类染色体的技术。方法应用原位引物标记（ＰＲＩＮＳ）在８例女性外周血淋巴细胞培养标本中检测了Ｘ、１８号染色体。结果在间期核及分裂相中均能特异地检测Ｘ、１８号染色体，Ｘ染色体的标记效率为８４％～９２％（平均８９％），１８号染色体为７３％～８７％（平均８４％）。标本经适当浓度的蛋白酶Ｋ处理后可以明显提高标记效率，增强标记信号。标记反应可利用原位ＰＣＲ仪进行，不到一小时即可完成。结论原位引物标记是一种快速、特异的染色体检测方法，利用这一技术将有可能在产前快速诊断染色体非整倍体。     

精神发育迟滞患儿的细胞遗传学研究

本室对１５６０例轻度精神发育迟滞患儿（魏氏智测法ＩＱ５１－８０）外周血染色体进行了分析，染色体异常２７例（占总数１．７％），染色体结构异常１８例，其中９号染色体臂间倒位患儿８名，约占染色体结构畸变病例总数的４４．４４％；染色体数目异常９例，均为性染色体数目异常，患儿的脆性表达频率平均为１８．４５±１１．７６％，显著高于正常对照组。结果提示９号染色体臂间倒位对精神发育的影响有待进一步探讨，脆性表达频率的增高对智力的影响不容忽视。     

胃癌基因表达的免疫组化研究

应用免疫组化方法对４２例胃癌和相位癌旁组织以及１１例良性胃病组织的ｐ５３、ｃ－ｍｙｃ和ｒａｓ基因编码产物进行检测。结果发现；３种基因在胃癌中的阳性率分别是３３．３％、２３．８％和４２．９％；在相应癌旁组织的阳性率依次分别是１６．７％、１１．９％和１６．７％；ｐ５３和ｃ－ｍｙｃ在在良性胃病组织中无表达，ｒａｓ基因在良性胃病组织中阳性率２．４％。     

Role of multicolor fluorescence in situ hybridization (FISH) in simultaneous detection of probe sets for chromosome 18, X and Y in uncultured amniotic fluid cells.

Major aneuploidies diagnosed prenatally involve the autosomes 13, 18, and 21, and sex chromosomes. Fluorescence in situ hybridization (FISH) allows rapid analysis of chromosome copy number in interphase cells. The purpose of this study was to evaluate the role of multicolor fluorescence in situ hybridization in simultaneous detection of probe sets for chromosome 18, X, and Y in uncultured amniotic fluid cells as a safer alternative method for aneuploidy detection prenatally. Fifty amniotic fluid samples were analyzed by FISH and standard cytogenetics. Mean time to obtain results was three days for fluorescence in situ hybridization and 20 days for karyotype. Fluorescence in situ hybridization was informative in 43 samples (86%), and within this group, two aneuploidies were correctly identified. This evaluation demonstrates that FISH with X, Y, and 18 alpha satellite DNA probes could accurately and rapidly detect aneuploidies involving these chromosomes and could be used in any prenatal clinical laboratory.     

Detection of numerical chromosomal abnormalities in malignant cells on body fluids by fluorescence in situ hybridization of interphase cell nuclei with chromosome-specific probes.

To detect numerical chromosomal abnormalities (NCA) in malignant cells on body fluids, Fluorescence in situ hybridization (FISH) technique was tested in clinical specimens from patients with metastatic disease. Directly labeled DNA probes specific for chromosomes 8, 12, X, and Y (Imagenetics, Naperville, IL) were used for in situ hybridization to interphase cell nuclei. Fifteen body fluids (BF) from various sites were studied. Based initially on the Papanicolaou-stained slides, there were seven malignant and eight benign samples. Blind analysis (200 cells/sample) showed that all benign samples had a normal number of chromosomes, whereas six of seven malignant samples showed different NCA comprising 5-60% of the cell population ranging from three to 10 chromosome signals per cell. We conclude that interphase cytogenetic cell analysis of BF by FISH is: (1) feasible and gives superior signals for detection of NCA, (2) helpful in detecting malignant cells, (3) relatively simple with a turnaround time of less than 24 hr. This method may have diagnostic and prognostic application in the study of the biologic behavior of malignant neoplasms.     

Multiplex interphase FISH as a screen for common aneuploidies in spontaneous abortions

BACKGROUND: A multiplex fluorescence in-situ hybridization (FISH) strategy using chromosome-specific probes for eight chromosomes as an initial screen for chromosome abnormalities in uncultured tissues from spontaneous abortions was evaluated. METHODS: Fifty-seven prefetal spontaneous abortions were studied by karyotyping cultured cells and using FISH on uncultured cells. Two probe sets were used, identifying chromosomes 13, 15, 16, 18, 21, 22, X and Y. RESULTS: Abnormalities were detected in 53% of cases by karyotyping, and 54% of cases by FISH. FISH detected an abnormality in four of five cases where cultures failed, and in two cases where maternal cells apparently overgrew the culture. FISH missed four trisomies not identifiable with the probe sets, and one trisomy because one probe set was unscorable. FISH using these probes identified 83% of all abnormalities detected by karyotyping. CONCLUSIONS: FISH can detect abnormalities in a significant proportion of cases where the culture fails to grow or is contaminated by maternal cell growth. Multiplex FISH as an initial screen, followed by culture and karyotyping in cases where no abnormality is detected, would identify a higher proportion of chromosome abnormalities in spontaneous abortion specimens than karyotype analysis alone.     

Incidence of chromosome numerical changes in multiple myeloma: fluorescence in situ hybridization analysis using 15 chromosome-specific probes.

The presence of complex karotypes with frequent numerical and structural abnormalities has been reported in 20 to 50% of multiple myeloma (MM) patients. This variability is mainly due to the difficulty of conventional cytogenetics to obtain tumor metaphases representative of all possible neoplastic clones in MM. To gain insight into the real incidence of numerical chromosome changes in MM we have studied by fluorescence in situ hybridization technique 15 different human chromosomes, 1, 3, 6, 7, 8, 9, 10, 11, 12, 13, 15, 17, 18, X, and Y, in a series of 52 MM patients. In all cases, the DNA index assessed by a propidium iodide/CD38 double-staining technique with flow cytometry was simultaneously investigated for correlation, with fluorescence in situ hybridization results. Additional aims of this study were 1) to analyze whether the abnormalities detected were common to all plasma cells or were present in only a subpopulation of tumor cells, 2) to explore changes caused by disease progression, and 3) to establish possible associations among the altered chromosomes. Although the overall incidence of numerical abnormalities was 67%, this frequency increased to 80% in the 41 cases in which 7 or more chromosomes were analyzed. Trisomies were significantly more common than monosomies (84% versus 16%). Chromosomes 9 and 15 were the most frequently altered (52% and 48% of cases, respectively), with all of their abnormalities corresponding to trisomies. The most frequent losses involved chromosomes 13 (26%) and X in females (32%). Other common numerical changes corresponded to chromosomes 1 (39%), 11 (37%), 6 (32%), 3 (31%), 18 (29%), 7 (28%), and 17 (22%). By contrast, chromosomes 8(13%), 10(8%), and 12(3%) were rarely altered. DNA aneuploidy by flow cytometry was detected in 67% of patients, and a high degree of correlation was observed between the DNA index obtained by flow cytometry and the chromosome index derived from fluorescence in situ hybridization studies, calculated according to two mathematical formulas (coefficient of correlation of 0.82 and 0.91 when at least 7 or 12 chromosomes were considered, respectively). The frequency of numeric chromosome aberrations was higher in those patients with progressive disease and, interestingly, trisomy of chromosome 8 was exclusively detected in this latter group of patients. Our study shows that, with the exception of chromosome 8, a possible marker of clonal evolution, the numeric chromosome changes are present in nearly all malignant plasma cells (r > 0.84). Finally, frequent associations between chromosomal aberrations were observed (ie, chromosomes 6, 7, 9, and 17; 7 and 15; and 11 and 17). By excluding them, it was found that two triple combinations of chromosome-specific probes, chromosomes 1 and 9 together with either chromosome 13 or 15, could be a useful marker for detection of residual disease, as it permits the identification of most MM patients displaying numerical changes.     

Sensitive detection of chromosome copy number aberrations in prostate cancer by fluorescence in situ hybridization.

The pattern of chromosomal aberrations and their significance in prostate cancer are poorly understood. We studied 23 prostate cancer and 10 benign prostatic hyperplasia (BPH) specimens by fluorescence in situ hybridization (FISH) using pericentromeric repeat-specific probes for 10 different chromosomes. The aims of the study were: 1) to compare the sensitivity of FISH and DNA flow cytometry in aneuploidy detection, 2) to determine which chromosome copy number changes are most common, and 3) which probe combinations would be most effective in aneuploidy diagnosis. Disaggregated tumor cells from formalin-fixed, paraffin-embedded tissues were pretreated with our newly developed method based on hot glycerol solution to improve probe penetration. All BPH specimens were diploid by DNA flow cytometry and showed no numerical chromosome aberrations by FISH. In prostate cancer, flow cytometry showed abnormal DNA content in 35% of cases, whereas 74% were abnormal by FISH. Aberrant copy number of chromosomes 8 (48% of cases), X (43% of cases), and 7 (39% of cases) were most common. Ninety-four percent of all aneuploid cases would have been detected with these three probes alone. Simple chromosome losses were uncommon but in DNA tetraploid tumors relative losses (trisomy or disomy) of several chromosomes were often found, suggesting progression of prostate cancer through tetraploidization followed by losses of selected chromosomes. In conclusion, our results indicate that FISH using three selected chromosome-specific probes is two to three times more sensitive than flow cytometric DNA content analysis in aneuploidy detection.     

Application of fluorescence in situ hybridization in hematological disorders.

In the present study, chromosome changes in bone marrow (BM) or peripheral blood (PB) cells from 13 patients with malignant hematologic disorders were analyzed by classical cytogenetic techniques (G-banding) and fluorescence in situ hybridization (FISH) procedures using centromere specific probes for chromosomes 1, 6, 7, 8, 9, 12, 18, 13/21, and X, and a DNA probe specific for the long arm of chromosome Y. The cytogenetic data obtained with G-banding were in accord with those obtained by FISH to metaphase chromosomes. Most significantly, FISH to interphase nuclei offered reliable results and in some cases provided important information concerning crucial chromosome anomalies which were not or could not be completely detected by analyzing metaphase chromosomes. Our results indicate that FISH could be clinically valuable in five major areas: 1) marker chromosome identification; 2) identification of trisomy consistent with certain specific hematological neoplasms; 3) clonal evaluation post observation of a single cell with trisomy; 4) clonal evaluation post-sex-mismatched bone marrow transplantation (BMT); and 5) residual disease detection following clinical remission.     

应用荧光原位杂交技术检测肺癌患者染色体数目的改变

目的应用荧光原位杂交(FISH)技术研究肺癌患者染色体数目改变及其意义。方法用人的全套染色体特异探针与肺癌患者中期染色体进行杂交检测。结果肺癌患者细胞为异倍体,染色体数目以亚二倍体居多,常见2、5、7、8、10、11、14和17号染色体增多及1、4号染色体的丢失。结论 FISH技术可检测肺癌患者染色体数目的改变,肺癌遗传学变异主要是2、5、7、8、10、11、14、17号染色体的增加,并对肺癌的发生、发展和预后有一定的提示作用。     

Chromosomal changes in prostate cancer: a fluorescence in situ hybridization study

The incidence of prostate cancer (PC) is increasing steadily with the aging population in Singapore. As the pattern of chromosomal aberrations in Asian men with PC is poorly understood, we investigated the numerical aberrations for chromosomes 7, 8, 11, and 17 by fluorescence in situ hybridization (FISH). FISH was performed on standard sections and tissue microarrays of 54 PC and 33 benign prostatic hyperplasia (BPH) specimens. Among the 54 PC specimens, FISH detected 44 cases as aneusomy and two as disomy and was unsuccessful for eight cases. Cytogenetic alterations of two or more chromosomes per tumor were detected in 33/46 (72%) PCs. The most frequent alteration was aneusomy of chromosome 8 detected in 34/46 (74%) cases followed by numerical aberrations in chromosome 7 (61%). Gain of 8q24, loss of chromosome 7, and gain of 11q13 were associated with higher Gleason score and were statistically significant. Gain of chromosome 7 was more common in locally advanced disease, while gain of chromosome 11q13 and chromosome 7 was more common in metastatic disease.     

Detection of genetic alterations in primary bladder carcinoma with dual-color and multiplex fluorescence in situ hybridization

Cytogenetic studies of bladder cancer have shown several nonrandom aberrations. Numerical aberrations of both sex chromosomes were investigated in 32 primary bladder tumors with bicolor fluorescence in situ hybridization (FISH). Loss of chromosome Y and overrepresentation of chromosome X were observed in subgroups of male patients. Chromosome X was represented normally in female patients. Two of the above primary bladder tumors, a transitional cell carcinoma (TCC) and an adenocarcinoma, were further analyzed with both multiplex FISH (24-color M-FISH) and G-banding. Both cases exhibited 1) common breakpoints on 5q11 approximately q12 and 15q24; 2) involvement of the pericentromeric area of chromosome 13; 3) structural abnormalities of chromosomes 8 and 17, with loss of material on the short arm; 4) structural abnormalities involving chromosome 11; and 5) loss of chromosome Y. The TCC case also exhibited structural abnormalities of chromosome 9, resulting in loss of 9q. The combined G-banding and M-FISH findings could help reveal regions potentially involved in bladder tumorigenesis.     

未培养羊水荧光原位杂交快速检测60例胎儿常见染色体数目异常

目的 应用细菌人工染色体(bacterial artificial chromosome,BAC)克隆自行制备荧光探针,对胎儿常见染色体数目异常(13,18,21,X,Y)行快速产前诊断.方法 利用中国医学遗传学国家重点实验室BAC库中相应染色体特异位点克隆,自行制备荧光探针.经外周血淋巴细胞染色体杂交验证后,用于胎儿未培养羊水的快速荧光原位杂交fluorescence in situ hybridization,FISH)检测,已检测60例羊水标本.结果 所有探针特异性均为100%,杂交成功率为97.86%;FISH结果与常规核型分析一致,检出21三体2例,18三体1例.有两例涉及其它染色体的结构异常未能检出.结论 自制探针用于未培养羊水快速FISH,使用标本量少、快速、简便,可有效检出上述5种染色体的数目异常,但本法不能检出其他染色体的数目异常和结构异常,其应用仍有一定局限性.