Biomedical subjects
Publications and source records attributed to B Perissel.
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.
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.
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.
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.
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.
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.
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.
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.
After a brief description of the automatic metaphase finding and karyotyping systems actually available, the authors describe an interactive method for chromosome analysis. The edges of each chromosome are delineated automatically. The use of 256 grey levels and 512 x 512 pixels allows the accurate classification. The result may be recorded on hard copy, videotape or disk. Present improvements of the Chromoscan include histograms, quantitative studies and the use of an expert system.
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Chromosomic alterations can be observed after the effect of a antimitotic amsidine on metaphases of cultured human lymphocytes: gaps, breaks, exchange figures, deletions, rings. Moreover we notice, among these cells, particular lines of big sized lymphoplasmocytoid cells, whose rate is abnormally high when compared to reference cultures.
The authors present a new interactive system for chromosome analysis. All the operations are checked by the biologist with the use of a "mouse". This system avoids the photographic work and the manual classification. Its main interest is to carry out quickly the karyotype after the slide preparation.
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.
In human lymphocytic metaphasis chromosomes sodium glycocholate induces a banding of chromatids and an intense apparently perichromatic labelling, reproducible on the same points of various chromosomal pairs. Arginine makes centromeric heterochromatin visible.
Six antitumoral drugs were tested in human lymphocytes cultures. Our observations pointed out specific abnormalities beside more usual chromosomal aberrations. These lesions are mainly telomeric and chromatidic fusions, chromomerisations and despiralizations. We present an interpretation of these abnormalities.
There have been conflicting studies of lymphocyte capping from patients with Duchenne Muscular Dystrophy. We have evaluated the proportion of capped fibroblasts of 10 patients with Duchenne Muscular Dystrophy. The results, compared with 15 normal controls, showed that the reduction of fibroblast capping is correlated to age patients. Additional studies, decreased number of retracted fibroblasts after colchicine incubation and ultrastructural observations, suggest that capping deficit would be the consequence of microtubular system alteration. This late phenomenon may be secondary to the primary membrane defect.
A patient with trisomy 9p in association with monosomy of the heterochromatic distal portion of the Y chromosome is reported. The rearrangement is probably due to malsegregation of a translocation 9p, Y and formation of an iso (9p). The phenotype of the patient is characteristic of trisomy 9p. There is a significant increase of GALT activity.