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Cytogenetic analysis of neuroblastoma
164
Abstracts 32
ANALYSISOF N-MYC ONCOGENE IN NEUROBLASTOMA CELLS FROM 53 PATIENTS. G. Wang1, V. Combaret2, G. Lenoir1, I. Philip2, T. Philip2, and M. C. Favrot2. 1International Agency for Research on Cancer, 150 cours A. Thomas, Lyon, 2Bone Marrow Transplant Unit, Centre L~on B6rard, 28 rue La~nnec, Lyon, France.
Southern blot analysis of neuroblastoma cells from 52 patients (58 samples) permitted to detect a N-myc amplification in 12 of the 52 patients and 13 of the 58 samples~ but a single copy of L and c-myc. Myc oncogene was analyzed either at diagnosis or relapse on largely contaminated BM samples [20 cases), on U.S. guided punctures (7 cases) or surgical biopsies (14 cases) of the primary tumors; it was also analyzed on 17 tumors taken after induction therapy during curative surgery. The quantification of the N-myc copies can be considered as accurate in pathological BM samples, U.S. guided punctures and surgical biopsies at diagnosis although the amplification of less than 10 copies could have been masked by the stroma reaction in stage III neuroblastoma, or in BM with less than 50% contamination. In contrast, tumors taken after therapy are histologically modified and only positive results must be considered as valid. In this series, the N-myc amplification in stage IV neuroblastoma over one year of age is rather low, regardless of the quality of the samples analyzed {10 of the 36 patients]. Amplification was detectable in only 2 of the 12 stage III neuroblastoma patients and none of the 3 patients balow one year of age, although relapses were observed in both groups.
33
CYTOGENETIC ANALYSIS OF NEUROBLASTOMA. lskra Petkovic, Melita Nakic, and Mladen Cepulid. Institute foi Mother and Child Health, Zagreb, Yugoslavia.
In this report the results of cytogenetic analysis in a boy with metastatic neuroblastoma are described. T. M. was a 14-month-old boy referred to the hospital in a poor general condition and with abdominal tumour. Incomplete surgical removal of the turnout mass was performed. Cytologic and pathohistologic analysis revealed neuroblastoma. Bone marrow aspiration showed infiltration of tumour cells with rosette formation. Radiation therapy was started but not continued because of poor physical condition of the patient, septicaemia, ascites and reduction of platelet count. Soon after the patient died. Direct cytogenetic analysis was performed on tumour pieces obtained from surgery. Cell suspension, was prepared by mechanical dissociation. After one hour of colchicine treatment at 37°C, the cells were treated with 0.075 M KCI for 15 min and fixed over night in 3:1 methanol-acetic acid. The chromosomes were stained by conventional Giemsa and modified trypsin.Giemsa method. A total of 53 cells were analysed while 28 were karyotypad and numerical and structural chromosome aberrations were identified. The modal chromosome number was 48. in all karyotyped cells chromosome Y and No. 18 were missing, while addition of No. 7 and 14 were identified. B~sides a small fragment, two rearranged No. 1 were observed, resulting in the monosomy for the distal segment of the short arm and trisomy for the long arm. In one near tetraploid cell the rearranged chromosomes were duplicated,
34
MHI-I-NBll, A NEW NEUROBLASTOMA CElL LINE. E. G6tter!, T. Pietsch 2, H. Riehmz, N. Blin2, and G. Kovacs~. 1Human Genetics, University of the Sear, Homburg; 2Pediatric Oncology and 3Pathology, Medical School, Hannover; F.R.G.
A continuous cell line was established from an explanted tumor biopsy obtained from a patient with advanced neuroblastoma. This cell line, MHH-NBll, retained most properties of the original immature tumor. It consisted of small, dense cells with scant cytoplasm and thin, long processes and it expressed neither GFAP nor S-100 protein but neuron-specific enolase and synaptophysin as judged by immunohistology. The karyotype (49 to 54 chromosomes; modal 52) showed a few structural aberrations, trisomy 2, 7, 8, 20, and a long HSR on the long arm of one of the chromosomes 13. Molecular hybridization in situ resulted in clusters of grains [26 of total 64) on 13qHSR with the N-myc (pNb-1} probe. Southern blots using dilution steps of genomic DNA suggest a 20-fold amplification of this oncogene.
Abstracts 32
ANALYSISOF N-MYC ONCOGENE IN NEUROBLASTOMA CELLS FROM 53 PATIENTS. G. Wang1, V. Combaret2, G. Lenoir1, I. Philip2, T. Philip2, and M. C. Favrot2. 1International Agency for Research on Cancer, 150 cours A. Thomas, Lyon, 2Bone Marrow Transplant Unit, Centre L~on B6rard, 28 rue La~nnec, Lyon, France.
Southern blot analysis of neuroblastoma cells from 52 patients (58 samples) permitted to detect a N-myc amplification in 12 of the 52 patients and 13 of the 58 samples~ but a single copy of L and c-myc. Myc oncogene was analyzed either at diagnosis or relapse on largely contaminated BM samples [20 cases), on U.S. guided punctures (7 cases) or surgical biopsies (14 cases) of the primary tumors; it was also analyzed on 17 tumors taken after induction therapy during curative surgery. The quantification of the N-myc copies can be considered as accurate in pathological BM samples, U.S. guided punctures and surgical biopsies at diagnosis although the amplification of less than 10 copies could have been masked by the stroma reaction in stage III neuroblastoma, or in BM with less than 50% contamination. In contrast, tumors taken after therapy are histologically modified and only positive results must be considered as valid. In this series, the N-myc amplification in stage IV neuroblastoma over one year of age is rather low, regardless of the quality of the samples analyzed {10 of the 36 patients]. Amplification was detectable in only 2 of the 12 stage III neuroblastoma patients and none of the 3 patients balow one year of age, although relapses were observed in both groups.
33
CYTOGENETIC ANALYSIS OF NEUROBLASTOMA. lskra Petkovic, Melita Nakic, and Mladen Cepulid. Institute foi Mother and Child Health, Zagreb, Yugoslavia.
In this report the results of cytogenetic analysis in a boy with metastatic neuroblastoma are described. T. M. was a 14-month-old boy referred to the hospital in a poor general condition and with abdominal tumour. Incomplete surgical removal of the turnout mass was performed. Cytologic and pathohistologic analysis revealed neuroblastoma. Bone marrow aspiration showed infiltration of tumour cells with rosette formation. Radiation therapy was started but not continued because of poor physical condition of the patient, septicaemia, ascites and reduction of platelet count. Soon after the patient died. Direct cytogenetic analysis was performed on tumour pieces obtained from surgery. Cell suspension, was prepared by mechanical dissociation. After one hour of colchicine treatment at 37°C, the cells were treated with 0.075 M KCI for 15 min and fixed over night in 3:1 methanol-acetic acid. The chromosomes were stained by conventional Giemsa and modified trypsin.Giemsa method. A total of 53 cells were analysed while 28 were karyotypad and numerical and structural chromosome aberrations were identified. The modal chromosome number was 48. in all karyotyped cells chromosome Y and No. 18 were missing, while addition of No. 7 and 14 were identified. B~sides a small fragment, two rearranged No. 1 were observed, resulting in the monosomy for the distal segment of the short arm and trisomy for the long arm. In one near tetraploid cell the rearranged chromosomes were duplicated,
34
MHI-I-NBll, A NEW NEUROBLASTOMA CElL LINE. E. G6tter!, T. Pietsch 2, H. Riehmz, N. Blin2, and G. Kovacs~. 1Human Genetics, University of the Sear, Homburg; 2Pediatric Oncology and 3Pathology, Medical School, Hannover; F.R.G.
A continuous cell line was established from an explanted tumor biopsy obtained from a patient with advanced neuroblastoma. This cell line, MHH-NBll, retained most properties of the original immature tumor. It consisted of small, dense cells with scant cytoplasm and thin, long processes and it expressed neither GFAP nor S-100 protein but neuron-specific enolase and synaptophysin as judged by immunohistology. The karyotype (49 to 54 chromosomes; modal 52) showed a few structural aberrations, trisomy 2, 7, 8, 20, and a long HSR on the long arm of one of the chromosomes 13. Molecular hybridization in situ resulted in clusters of grains [26 of total 64) on 13qHSR with the N-myc (pNb-1} probe. Southern blots using dilution steps of genomic DNA suggest a 20-fold amplification of this oncogene.