Numerical abnormalities of chromosome 7 in human prostate cancer detected by fluorescence <i>in situ</i> hybridization (FISH) on paraffin‐embedded tissue sections with centromere‐specific dna probes

Zitzelsberger, Horst; Szűcs, Sándor; Weier, Heinz‐Ulrich G.; Lehmann, Lars; Braselmann, Herbert; Enders, Susanne; Schilling, A.; Breul, Jürgen; Höfler, Heinz; Bauchinger, M.

doi:10.1002/path.1711720407

Cited by 59 publications

(36 citation statements)

References 20 publications

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“…However, it has to be kept in mind that evaluation of signals is hampered in general by cutting artifacts based on truncation of nuclei. This effect leads to a decrease in signal number, with two signals in approximately 80 -85% of the nuclei instead of 95% normally found in preparations of intact nuclei (6,8,15). In particular, detection of monosomy may be difficult, with thresholds at nearly 30%.…”

Section: Discussionmentioning

confidence: 99%

Detection of Chromosomal Aberrations in Well-Differentiated Hepatocellular Carcinoma by Bright-Field In Situ Hybridization

et al. 2002

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Differentiation between well-differentiated hepatocellular carcinoma (HCC) and nonmalignant lesions with increased cellular proliferation may be difficult in needle biopsies. Based on recurrent chromosome aberrations known for HCC, we developed a nonfluorescent in situ hybridization technique that allows combination with morphological analysis in bright-field microscopy. Fourteen biopsies of HCC and 31 samples of regenerative nodules (n ‫؍‬ 10), chronic hepatitis (n ‫؍‬ 10), fibrosis or cirrhosis of unknown origin (n ‫؍‬ 5), focal nodular hyperplasia (n ‫؍‬ 2), primary biliary cirrhosis (n ‫؍‬ 2), steatosis (n ‫؍‬ 1), and adenomatous hyperplasia (n ‫؍‬ 1) were analyzed with probes specific for the centromeric regions of chromosomes 1, 6, 7, and 8. After microwave pretreatment and in situ hybridization, signals were detected using a tyraminebased system and AEC as substrate. Evaluation of signals was done by conventional bright-field microscopy. Using this approach, aberrant counts were seen for at least one chromosome in 12/14 cases of HCC. In contrast, none of the nonmalignant lesions revealed aberrant counts for any of the chromosomes analyzed. In conclusion, this new combination of in situ hybridization and tyramine amplification allows fast and reliable evaluation of chromosome aberrations in a histomorphological context similar to paraffin immunohistochemistry. Registration of imbalances contributes to a reliable differentiation between malignant and nonmalignant lesions of the liver. The fast and reliable differentiation between malignant and nonmalignant tumorlike lesions of the liver is of the utmost importance for the further treatment and surgical procedure of the patients. However, even for the experienced pathologist, in some cases, determining the correct diagnosis may be extremely difficult and uncertain, particularly if only small biopsies are available as the main source for histological examination. In particular, differentiation between well-differentiated hepatocellular carcinoma (HCC) and benign alterations may be impossible based on morphologic criteria alone (1). Detection of chromosomal aberrations within questionable tumors could contribute toward solving this problem. Because of the time and effort required for conventional cytogenetics, this technique is usually not appropriate. The recently developed comparative genomic hybridization (CGH) has revealed promising results (2,4,11,13,14), but it is also of limited value because of the duration and amount of tissue required. By contrast, fluorescence in situ hybridization (FISH) yields results that are evaluable by simple counting of fluorescence signals in conventional biopsies (12). A major limitation of this approach is, however, that most pathologists are not familiar with the morphology of the tumor in histological sections counterstained with fluorescent dyes. Furthermore, many histomorphological details remain undetected in the dark field required for evaluation. In addition, epifluorescence microscopy is based on expensive technica...

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Section: Discussionmentioning

confidence: 99%

Detection of Chromosomal Aberrations in Well-Differentiated Hepatocellular Carcinoma by Bright-Field In Situ Hybridization

et al. 2002

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“…FISH reactions were prepared with slight modifications of established protocols (Zitzelsberger et al, 1994;Aubele et al, 1997). The predigestion stages included: 70% formic acid for 15 min, incubation in 1 M NaSCN at 80°C, for 10 min, and Pronase digestion.…”

Section: Slide Preparation and Hybridizationmentioning

confidence: 99%

“…One microlitre of CEP 1 probe (Vysis, Germany), 2 µl of purified water and 7 µl of CEP hybridization buffer was used in hybridization. Post-hybridization washing was performed at 43°C as previously described (Zitzelsberger et al, 1994). Nuclei were counterstained with 1 µg ml -1 4′,6-diamidino-2-phenylindole (DAPI) in antifade solution.…”

Section: Slide Preparation and Hybridizationmentioning

confidence: 99%

Increasing chromosome 1 copy number parallels histological progression in breast carcinogenesis

Cummings

Aubele²,

Mattis

et al. 2000

Br J Cancer

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Summary Chromosome 1 copy number in the benign breast lesions hyperplasia and atypical duct hyperplasia (ADH) was investigated using fluorescence in situ hybridization on paraffin sections. Progression of chromosome 1 changes occurring in parallel with histological progression from normal through hyperplasia and ADH to ductal carcinoma in situ (DCIS) and invasive carcinoma was also assessed, both overall and within individual patients. The mean signal number for normal cells was 1.14, while that for hyperplasia was 1.56 and ADH was 1.5, while values for DCIS of 1.95 and invasive duct carcinoma of 1.79, were higher (P < 0.001). Six of the seven cases also showed a significant trend towards an increasing proportion of cells with greater than 2 signals per nucleus occurring with histological progression (P < 0.001). These results support the concept that benign proliferative breast disease is a biological precursor of in-situ and invasive ductal carcinoma, the early histological changes possibly indicating a field effect with further genetic changes required for the development of a malignant phenotype.

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“…While the application of FISH to studies of the association between DNA aneuploidy and prognosis has attracted attention, no numerical aberration of any chromosome which might be specific for prostate cancer has so far been established. Furthermore, it is unclear whether the numerical aberrations of individual chromosomes that have been detected (Takahashi et al, 1994;Visakorpi et al, 1994;Zitzelsberger et al, 1994) have pathological and/or clinical significance. However, these previous studies of prostate cancer analysed only a few cases or used limited numbers of cx-satellite DNA probes.…”

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confidence: 99%

Interphase cytogenetics of prostate cancer: fluorescence in situ hybridisation (FISH) analysis of Japanese cases

Mizunuma

Shiraishi²,

Yatani³

et al. 1996

Br J Cancer

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Summary No numerical aberration of chromosomes that might be specific for prostate cancer has so far been established. We used fluorescence in situ hybridisation (FISH) with centromere-specific probes for chromosomes 7, 8, 17, X and Y to establish the distribution of centromere copy numbers in frozen-stored or freshly prepared samples of benign prostate hypertrophy (BPH) and to detect numerical aberrations of these chromosomes in 28 prostate cancers from Japanese men. There was no significant difference in the data of centromere copy numbers between fresh and frozen-stored tissue. The most common aberration in prostate cancers was a gain of chromosome 8 (57%), with numerical aberration of chromosome 7 being the second most frequent anomaly (50%). Numerical aberration of chromosome 7 is most significantly associated with a higher Gleason score (GS) (P<0.005) or with lymph node metastasis (P<0.001). Numerical aberration of several chromosomes, including chromosomes 7 and/or 8, was common in aggressive prostate cancers. Loss of chromosome Y was detected in only 4% of cases. FISH analysis thus proved to be a useful method for detecting numerical aberrations of individual chromosomes, with application to touch preparations of frozenstored tissue having the advantage of exact sampling of cancer foci. The results suggest that numerical aberration of chromosome 7 is associated with aggressive tumour behaviour and poor prognosis of patients with prostate cancer. The association between genetic change and chromosomal abnormality should be studied in detail.Keywords: interphase cytogenetics; fluorescence in situ hybridization; prostate cancer Fluorescence in situ hybridisation (FISH) has been used to hybridise specific nucleic acid sequences with complemental DNA fragments, revealing their location on chromosomes and their copy numbers (Trask et al., 1990). Compared with conventional metaphase cytogenetics or karyotyping analysis, FISH allows more rapid enumeration of specific chromosomes and detection of chromosomal alterations even in interphase nuclei (interphase cytogenetics) as well as in metaphase nuclei (Cremer et al., 1986;Pinkel et al., 1986). This technique does not always require tissue culturing and can be applied to solid tumours (Persons et al., 1993;Devilee et al., 1988) and even formalin-fixed, paraffin-embedded tissues (Micale et al., 1993;Persons et al., 1994). FISH has been demonstrated to be more sensitive than flow cytometry (FCM) for detecting aneuploidy (Takahashi et al., 1994;Visakorpi et at., 1994), and the methodology can detect numerical aberrations of individual chromosomes at levels impossible with FCM and image cytometry (ICM). FCM also has limitations regarding minor quantitative DNA changes (Hopman et al., 1990). While the application of FISH to studies of the association between DNA aneuploidy and prognosis has attracted attention, no numerical aberration of any chromosome which might be specific for prostate cancer has so far been established. Furthermore, it is unclear whether the numeri...

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Numerical abnormalities of chromosome 7 in human prostate cancer detected by fluorescence in situ hybridization (FISH) on paraffin‐embedded tissue sections with centromere‐specific dna probes

Cited by 59 publications

References 20 publications

Detection of Chromosomal Aberrations in Well-Differentiated Hepatocellular Carcinoma by Bright-Field In Situ Hybridization

Detection of Chromosomal Aberrations in Well-Differentiated Hepatocellular Carcinoma by Bright-Field In Situ Hybridization

Increasing chromosome 1 copy number parallels histological progression in breast carcinogenesis

Interphase cytogenetics of prostate cancer: fluorescence in situ hybridisation (FISH) analysis of Japanese cases

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