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Cytogenetic analysis of epithelial pancreatic neoplasms
Abstracts
A21
C Y T O G E N E T I C ANALYSIS OF E P I T H E L I A L PANCREATIC NEOPLASMS E Long, J. Cbou, L. Morsberger, T. Ellingham, R. Hmban, C. Yco, C. Griffin Johns Hopkins School of Medicine, Depts. of Pathology and Surgery. Baltimore, MD 21287 Adenocarcinoma is clinically the most significant pancreatic neoplasm, leading to death in the majority of patients in a short time. Other less common neoplasms involving pancreatic epithelial cells include serous cystadenomas, mucinons cystadenocarciunmas, papillary and cystic neoplasms and papillary oncecytic neoplasms. Virtually nothing is known about somatic genetic changes occurring in these tumors. We have analyzed twelve tumors: six serous cystadeoomas, three mucinous cystudenocarcinomas, two solid and cystic neoplasms and one papillary oncocytic tumor. All twelve specimens were surgically resected and were processed using short term tissue culture methods for solid tumors. Of the six serous cystadenomas, three had normal karyotypes, one had an inadequate number of metaphases for analysis and two were abnormal. The clonal abnormalities included a probable deft 10) and add(22)(q 13) in one tumor and +X, -Y, +7 in another tumor. Of the three mucinons cystadenocarcinomas one had a normal karyotype, one did not grow and one had an unidentified ring chromosome in addition to a +X. The two solid and cystic tumors and the single papillary oncecytic tumor had only normal karyotypes. These results contrast with those obtained when we karyotyped over 100 ductal adenocarcinomas of the pancreas. The ductal adenocarcinomas had very complex karyotypes with many structural and numerical alterations. The difference between the two groups of tumors emphasizes the low malignant potential of the generally benign cystic tumors of the pancreas and contrasts them with adenocarcinomas of the pancreas.
A22
C O ~ A R I S O N OF FLUORESCENCE IN-SITU HYBRIDIZATION (FISH) AND BLADDER TUMOR A N T I G E N TEST (BTA®) IN BLADDER CANCER: A PILOT STUDY TS McConnell, AY Smith, LA Haines, Jr, T Cushing, ED Crawford, M Sinvak. Departments of Pathology anti Surgery, University of New Mexico Health Sciences Center, Albuquerque, NM, and Southwest Oncology Group (SWOG) [Geniroarinary Committee and Cytogenetics Committee], San Antonio, TX. As part of a larger SWOG multi-institution pilot study using FISH probes in ceils from bladder washings, we utilized bladder washings from 13 UNM patients to compare this technique with a latex agglutination assay for basement membrane complexes (BTA® Bard® Urological, Covington, Ga). Cell fixation and slide preparation were performed using Cytlyt®, PreservCyt® and ThinPrel~ slide processor (CYTYC® Corporation, Marlborough, MA). Repeat-sequence DNA probes for chromosomes 1, 7, 8, 9, 17 and Y were performed using the Vysis® lnc (Downers Grove, IL) Spectrum CEP@ methods. Bladder washings were obtained from patients with known transitional cell carcinoma of the urinary bladder or upper genitourinary system (positive cytology or surgical pathology) by a standard barbotage technique. All 6 FISH probes were suecessfuliy analyzed in ten patient samples, while three samples had only chromosomes 8 and 17 analyzed. Appropriate controls were analyzed with each run. Abnormal FISH results were found in six samples and abnormal BTA® results were found in seven samples. There was concordance between the two tests in eight patients, discordance in four, and no findings in one. In one analysis, 9% of signals were yellow, indicating probable fusion and perhaps chromosome translocatirm (we believe this figure is too high to be explained by somatic pairing). We conclude that using both these tests yields more information than either one alone, and that further comparisons are warranted.
161
A23
AN AMPLIFIED G E N E ENCODES T H E TRANSLATION INITIATION FACTOR eIF-4GAMMA W H I C H INDUCES AN IMMUNE RESPONSE IN A PATIENT W I T H SQUAMOUS CELL LUNG CARCINOMA E Meese 1, D Heckel t , M Pfreundschuh 2, G Sybrecht3, and N Brass 1 IDepartment of Human Genetics, Ban 68, Medical School, 2Department of Internal Medicine 1, University Hospital, 3Department of Internal Medicine V, University Hospital, University of Saarland, D-66421 Homburg/Saar, Germany Amplification of cellular oncogenes is an important mechanism of altered gene expression in human cancers. Using comparative genomic hybridization we recently identified an amplification at 3q26.l-q26.3 in 30% of squamous cell carcinoma of the lung. In this study, we have combined an immunological and molecular genetic approach to analyze squamous cell lung carcinoma. To identify beth amplified and tumor relevant genes, we generated a eDNA expression library from a tumor with the 3q26.l-q26.3 amplification and hybridized the expressed recombinant polypeptides with the autulogons serum. Out of 400 000 eDNA clones we identified 17 antigens which induce an immune response in patient with squamous cell lung carcinoma. While most clones represent individual antigens sequenCe analysis revealed that four of the 17 cDNAs are identical with the eukaryotic translation initiation factor (eIF)-4gamma reCently assigned on 3q. We demonstrated that the gene for elF-4gamma was amplified within 3q26.1-q26.3 in independent squamous cell lung carcinomas, eIF,-4gamma is likely to play a crucial role in the development of squamons cell lung carcinoma.
A24
MULTIPLE G E N E T I C A B N O R M A L r r l E S INCLUDING EVIDENCE OF CHROMOSOME I I Q I 3 R E A R R A N G E M E N T ~ IN PITUITARY ADENOMAS BY COMPARATIVE GENOMIC HYBRIDIZATION Metz~er A.K., Mohapatra G., Minn Y.A., Bollen A.W., Wilson C.B., Feuerstein B.G.; Departments of Lab Medicine and Neurosurgery, University of California, San Francisco, CA 94143-0808 Pituitary adenomas account for approximately 10% of intracranial neoplasms, but relatively few genetic abnormalities have been detected in these tumors. Previously used methods are hampered by the inability to examine the entire tumor gnnome and may be insensitive to some typos of genetic abnormalities. We used comparative genomic hybridization to study 40 primary and 14 recurrent adenomas from 53 patients. Total genomic DNA was extracted from frozen tumor specimens and normal leukocytes and nick translated with FITC-dUTP and Texas Red-dUTP, respectively. Labeled tumor and control DNA samples were denatured, hybridized to normal metaphase chromosomes, and analyzed with a quantitative image processing system. A change in the ratio of green to red signal def'med a copy number aberration (CNA). CNAs were detected in 29•54 (54%) adenomas. Whole chromosome C-'NAswere common (103/146=71% of CNAs) and often multiple. The average number of aberrations was 3.9 (79120) for primary and 6.8 (61/9) for recurrent tumors. Loss of all chromosome 11 material except for the llq13 region occurred in 4•53 (8%) patients. This was the only genetic aberration affecting a chromosomal subregion in this study and was the sole abnormality in 3/4 cases. We conclude that genetic aberrations are more common in pituitary adenomas than previously reported and that rearrangement of the 11ql 3 region may affect an oncogene important in the initiation or progression of these tumors. Supported by CA13525, 61147, 64898, and Vysis, Inc.
A21
C Y T O G E N E T I C ANALYSIS OF E P I T H E L I A L PANCREATIC NEOPLASMS E Long, J. Cbou, L. Morsberger, T. Ellingham, R. Hmban, C. Yco, C. Griffin Johns Hopkins School of Medicine, Depts. of Pathology and Surgery. Baltimore, MD 21287 Adenocarcinoma is clinically the most significant pancreatic neoplasm, leading to death in the majority of patients in a short time. Other less common neoplasms involving pancreatic epithelial cells include serous cystadenomas, mucinons cystadenocarciunmas, papillary and cystic neoplasms and papillary oncecytic neoplasms. Virtually nothing is known about somatic genetic changes occurring in these tumors. We have analyzed twelve tumors: six serous cystadeoomas, three mucinous cystudenocarcinomas, two solid and cystic neoplasms and one papillary oncocytic tumor. All twelve specimens were surgically resected and were processed using short term tissue culture methods for solid tumors. Of the six serous cystadenomas, three had normal karyotypes, one had an inadequate number of metaphases for analysis and two were abnormal. The clonal abnormalities included a probable deft 10) and add(22)(q 13) in one tumor and +X, -Y, +7 in another tumor. Of the three mucinons cystadenocarcinomas one had a normal karyotype, one did not grow and one had an unidentified ring chromosome in addition to a +X. The two solid and cystic tumors and the single papillary oncecytic tumor had only normal karyotypes. These results contrast with those obtained when we karyotyped over 100 ductal adenocarcinomas of the pancreas. The ductal adenocarcinomas had very complex karyotypes with many structural and numerical alterations. The difference between the two groups of tumors emphasizes the low malignant potential of the generally benign cystic tumors of the pancreas and contrasts them with adenocarcinomas of the pancreas.
A22
C O ~ A R I S O N OF FLUORESCENCE IN-SITU HYBRIDIZATION (FISH) AND BLADDER TUMOR A N T I G E N TEST (BTA®) IN BLADDER CANCER: A PILOT STUDY TS McConnell, AY Smith, LA Haines, Jr, T Cushing, ED Crawford, M Sinvak. Departments of Pathology anti Surgery, University of New Mexico Health Sciences Center, Albuquerque, NM, and Southwest Oncology Group (SWOG) [Geniroarinary Committee and Cytogenetics Committee], San Antonio, TX. As part of a larger SWOG multi-institution pilot study using FISH probes in ceils from bladder washings, we utilized bladder washings from 13 UNM patients to compare this technique with a latex agglutination assay for basement membrane complexes (BTA® Bard® Urological, Covington, Ga). Cell fixation and slide preparation were performed using Cytlyt®, PreservCyt® and ThinPrel~ slide processor (CYTYC® Corporation, Marlborough, MA). Repeat-sequence DNA probes for chromosomes 1, 7, 8, 9, 17 and Y were performed using the Vysis® lnc (Downers Grove, IL) Spectrum CEP@ methods. Bladder washings were obtained from patients with known transitional cell carcinoma of the urinary bladder or upper genitourinary system (positive cytology or surgical pathology) by a standard barbotage technique. All 6 FISH probes were suecessfuliy analyzed in ten patient samples, while three samples had only chromosomes 8 and 17 analyzed. Appropriate controls were analyzed with each run. Abnormal FISH results were found in six samples and abnormal BTA® results were found in seven samples. There was concordance between the two tests in eight patients, discordance in four, and no findings in one. In one analysis, 9% of signals were yellow, indicating probable fusion and perhaps chromosome translocatirm (we believe this figure is too high to be explained by somatic pairing). We conclude that using both these tests yields more information than either one alone, and that further comparisons are warranted.
161
A23
AN AMPLIFIED G E N E ENCODES T H E TRANSLATION INITIATION FACTOR eIF-4GAMMA W H I C H INDUCES AN IMMUNE RESPONSE IN A PATIENT W I T H SQUAMOUS CELL LUNG CARCINOMA E Meese 1, D Heckel t , M Pfreundschuh 2, G Sybrecht3, and N Brass 1 IDepartment of Human Genetics, Ban 68, Medical School, 2Department of Internal Medicine 1, University Hospital, 3Department of Internal Medicine V, University Hospital, University of Saarland, D-66421 Homburg/Saar, Germany Amplification of cellular oncogenes is an important mechanism of altered gene expression in human cancers. Using comparative genomic hybridization we recently identified an amplification at 3q26.l-q26.3 in 30% of squamous cell carcinoma of the lung. In this study, we have combined an immunological and molecular genetic approach to analyze squamous cell lung carcinoma. To identify beth amplified and tumor relevant genes, we generated a eDNA expression library from a tumor with the 3q26.l-q26.3 amplification and hybridized the expressed recombinant polypeptides with the autulogons serum. Out of 400 000 eDNA clones we identified 17 antigens which induce an immune response in patient with squamous cell lung carcinoma. While most clones represent individual antigens sequenCe analysis revealed that four of the 17 cDNAs are identical with the eukaryotic translation initiation factor (eIF)-4gamma reCently assigned on 3q. We demonstrated that the gene for elF-4gamma was amplified within 3q26.1-q26.3 in independent squamous cell lung carcinomas, eIF,-4gamma is likely to play a crucial role in the development of squamons cell lung carcinoma.
A24
MULTIPLE G E N E T I C A B N O R M A L r r l E S INCLUDING EVIDENCE OF CHROMOSOME I I Q I 3 R E A R R A N G E M E N T ~ IN PITUITARY ADENOMAS BY COMPARATIVE GENOMIC HYBRIDIZATION Metz~er A.K., Mohapatra G., Minn Y.A., Bollen A.W., Wilson C.B., Feuerstein B.G.; Departments of Lab Medicine and Neurosurgery, University of California, San Francisco, CA 94143-0808 Pituitary adenomas account for approximately 10% of intracranial neoplasms, but relatively few genetic abnormalities have been detected in these tumors. Previously used methods are hampered by the inability to examine the entire tumor gnnome and may be insensitive to some typos of genetic abnormalities. We used comparative genomic hybridization to study 40 primary and 14 recurrent adenomas from 53 patients. Total genomic DNA was extracted from frozen tumor specimens and normal leukocytes and nick translated with FITC-dUTP and Texas Red-dUTP, respectively. Labeled tumor and control DNA samples were denatured, hybridized to normal metaphase chromosomes, and analyzed with a quantitative image processing system. A change in the ratio of green to red signal def'med a copy number aberration (CNA). CNAs were detected in 29•54 (54%) adenomas. Whole chromosome C-'NAswere common (103/146=71% of CNAs) and often multiple. The average number of aberrations was 3.9 (79120) for primary and 6.8 (61/9) for recurrent tumors. Loss of all chromosome 11 material except for the llq13 region occurred in 4•53 (8%) patients. This was the only genetic aberration affecting a chromosomal subregion in this study and was the sole abnormality in 3/4 cases. We conclude that genetic aberrations are more common in pituitary adenomas than previously reported and that rearrangement of the 11ql 3 region may affect an oncogene important in the initiation or progression of these tumors. Supported by CA13525, 61147, 64898, and Vysis, Inc.