[Prognosis of chronic lymphocytic leukemia: focus on recent biomarkers]. / Pronostic de la leucémie lymphoïde chronique : mise au point sur les facteurs biologiques récents.

Early stages of chronic lymphocytic leukemia (CLL), which are the most frequent at diagnosis, have an extremely variable individual prognosis, as some patients remain stable for years whereas others develop aggressive forms of the disease less or more rapidly. Individual prognosis evaluation of early stages of CLL is then a challenge for physicians; also clinico-hematological stages are still the evaluation basis, numerous biological markers are helpful in providing independent information on patient prognosis. It is useful to distinguish the classical prognosis factors, described in the 1980s, and the recent markers described from the end of the 1990s, which are widely validated for certain, whereas for others further investigations are needed to confirm their prognostic impact. We propose to detail in this review these new prognostic factors of CLL, especially the different serum markers, cytogenetical abnormalities of pathologic lymphocytes, mutational status of the immunoglobulin genes (IgVH) and CD38 and ZAP-70 expression.

An unusual familial chromosome 9 "variant" with variable phenotype: characterization by CGH analysis.

Heterochromatin confined to pericentromeric and secondary constriction regions plays a major role in morphological variation of chromosome 9, because of its size and affinity for pericentric inversion. We report on a 6-year-old boy with growth and language delay, minor facial anomalies and unusual chromosome 9 variant with an extra-band in the centromeric region on the conventional karyotype. Subsequent analysis by FISH and CGH identified this variant as a dicentric chromosome 9 with a duplication of the 9p12-q21 region. An identical chromosome 9 variant was found in the mild language retarded brother and in the phenotypically normal father and grandfather. The presumed mechanism accounting for the phenotypic discordance observed in this family and the usefulness of CGH in characterization of such variants are discussed. To our knowledge, this is the first investigation of an unusual chromosome 9 variant by CGH.

Cytogenetic characterization of a bladder carcinoma cell line by fluorescence in situ hybridization.

A bladder carcinoma cell line, RT112/84, has been cytogenetically characterized by classical techniques and fluorescence in situ hybridization. The RHG banding and FISH analysis revealed a mixture of two clones and multiple chromosome rearrangements.

Rehybridization on metaphases studied previously by FISH. An approach to analyze chromosome aberrations.

A new protocol for fluorescence in situ rehybridization is described. Biotin-labeled chromosome-specific DNA probes were hybridized onto metaphases which previously had been studied by FISH. This method makes it possible to reexamine the same metaphase spreads with different DNA probes. It allows for precise characterization of cytogenetic translocations and for detection of multiple aberrations presented in a karyotype. It is especially useful in cases where a limited number of cytogenetic preparations are available.

Translocation t(3;20) associated with thrombocythemia in Ph-positive CML.

A patient with Philadelphia (Ph) chromosome positive chronic myelocytic leukemia is described who also developed an abnormality of chromosome #3, i.e., t(3;20)(p21;p13), in blast crisis. This abnormality may be connected with the advent thrombocythemia. The disease was a thrombopenia in the initial phase.

Karyotype and FISH analysis of a newly established cell line derived from a human bladder carcinoma.

A new human malignant urologic cell line was established in vitro from a moderately differentiated transitional cell carcinoma of the bladder and cytogenetically characterized. Repeated chromosome analyses of the cell line using conventional RHG and GTG banding and non-radioactive in situ hybridization showed a stable karyotype with a modal number of 48 and chromosomal rearrangements, some of which have not been previously described. Numerical deviation included three trisomies (+7, +8, +9) and one nullisomy (-19, -19). Structural changes involved a balanced translocation (1;5)(q12;q12), an isochromosome 3q, a 14p+, and two markers. Fluorescence in situ hybridization (FISH), using biotin-labeled alpha satellite probes for chromosome 9 or painting for chromosomes 1 and 8, applied to interphase nuclei or metaphases showed similar results to those found by conventional cytogenetic study. This cell line may be an interesting model for fuller characterization by molecular biology studies and for testing anti-cancer drugs in vitro.

Nonrandom changes of chromosome 10 in bladder cancer. Detection by FISH to interphase nuclei.

Fluorescence in situ hybridization (FISH) to interphase nuclei has been a valuable method for examining the chromosome copies in tumor cells in clinical practice. Twelve cases of transitional cell carcinoma (TCC) of the bladder were investigated with a biotin-labeled repetitive DNA probe to detect numerical aberrations of chromosome 10 in interphase nuclei. The cells containing one fluorescent signal were screened in two of seven non-invasive tumors and in four of five invasive tumors. Two patients presented two FISH spots of different sizes. More than two signals were seen in one invasive tumor. The findings suggest that partial or complete loss of a chromosome 10 is a nonrandom aberration in bladder cancer.

Structural and numerical aberrations of chromosome 22 in a case of follicular variant of papillary thyroid carcinoma revealed by conventional and molecular cytogenetics.

This study reports a case of papillary carcinoma with vesicular components showing multiclonal aberrations of chromosome 22 as revealed by RHG-banding cytogenetics and by fluorescence in situ hybridization (FISH; whole chromosome 22 and BCR-ABL-specific locus probes, multi-FISH). Four clones with chromosome 22 changes as the sole abnormality were seen. The main abnormal clone lacked the whole chromosome 22. A del(22)(q11) was observed in a second group of cells. The third clone had an idic(22). Finally, FISH revealed a fourth abnormal cell population with a der(17)t(?17;22). Some of these chromosome 22 alterations have been described in other solid tumors such as meningiomas and neurinomas, suggesting a common genetic pathway of tumor progression occurring in a multistep process. Chromosome 22 changes do not seem to be involved in pure papillary thyroid tumors and therefore could be related to the maintenance of a follicular-type histological pattern.

Chromosome analysis by image processing in a computerized environment. Clinical applications.

Dealing with a routine regional cytogenetic activity, we have developed and adapted to clinical work a semi automatic karyotyping machine. Attempts for an accurate automated chromosome classification using a neural network have led to partial results. A specific adaptation to cancer cytogenetics is under development (determination of the modal number, translocations analysis with densitometric curves, automatic identification of markers). A specific program allows quantification of chromosome labelling with radioactive probes. Exchanges of digitized karyotypes are feasible with labs using automated karyotyping machines. A local network connects several karyotyping and metaphase finding stations. Guidelines for an international data bank concerning abnormal chromosome images have been elaborated. On the other hand the ISH techniques have been applied to the following topics: identification of human chromosome aberrations in amniotic and chorionic cells, chromosome studies of human gametes and embryos (including sex determination), identification of markers in cancer cells.

Interphase cytogenetic studies of bladder cancer.

Transitional cell carcinomas of human urinary bladder were studied by interphase fluorescence in situ hybridization (FISH). With current hybridization to isolated nuclei, 26 tumors were investigated and nonrandom +7, -9 and -10 were identified. Monosomy 11, tetraploidies and polyploidies were detected in invasive and poor-differentiated tumors. Hybridization on frozen sections offers another means of analysing surgical samples. FISH to vesical washings can be applied to monitor tumor progression. Hybridizations on paraffine sections and on tissues previously stored in liquid nitrogen allow retrospective studies of the archived materials. Our data suggest that the interphase FISH can become a powerful tool for cytogenetic studies of bladder cancer.

[Chromosomal analysis of bladder tumors. Technical aspects, anatomoclinical correlations and perspectives. Report of 18 cases]. / Analyse chromosomique des tumeurs de vessie. Aspects techniques, corrélations anatomocliniques et perspectives. A propos de 18 cas.

The course of bladder tumours is difficult to predict. The most reliable prognostic factor at the present time is histological grade. Cytogenetic subclasses of bladder tumours can be distinguished on the basis of the demonstration of karyotype anomalies in bladder tumour cells. Eighteen patients underwent cytogenetic examination of their bladder tumour and were followed for an average of 35 months. Multivariate analysis of the clinical and laboratory parameters studied revealed the importance of age and the absence of trisomy 7 in the tumour on patient survival. The presence of trisomy 7 in a bladder tumour may therefore constitute a factor of poor prognosis. This hypothesis needs to be confirmed by further studies in larger populations. The search for this anomaly can be performed by fluorescent in situ chromosomal hybridisation, a technique which transforms cytogenetics from an experimental procedure into a routine complementary investigation. These techniques can be performed on urine samples, suggesting the possibility of their application to screening or follow-up of bladder tumours.

Simultaneous in situ hybridization with biotin-labeled centromeric and library DNA probes: a useful method for identifying translocations.

A method of in situ hybridization is described for rapid characterization of cytogenetical translocations. In the same experimental procedure, biotinylated centromeric and 'painting' DNA probes were used in combination. Signals of the double-target hybridization were visualized by only one fluorescein. This technique also permits a simultaneous detection of multiple unrelated aberrations involving several chromosomes.