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The recurrent chromosome changes in 29 squamous cell carcinomas of the head and neck region
Abstracts
147
LINEAGE-REBTRICTED CKROMOSOME U E R R A TIOH8 INDICATE CELL OF ORIGIN IN BIPHA8IC 8SLID TOMOR8. Fletcher JA. Departments of Pathology and Pediatrics, H a r v a r d M e d i c a l S c h o o l , B o s t o n , MA. 25
Many solid tumors contain a single welldefined neoplastic cell population. Others contain two or more histologically distinct cell lineages that might potentially be neoplastic. Several approaches can be used to determine the cell lineages responsible for clone1 chromosome aberrations in solid tumors. These approaches include: 1) selective tissue culture of different cell populations prior to metaphase cell h a r v e s t s ; 2) i m m u n o h i s t o c h e m i c a l staining of m e t a p h a s e cells; 3) i m m u n o h i s t o c h e m i c a l staining coupled with chromosomal in situ hybridization in interphase cells; a n d 4) chromosomal in s i t u h y b r i d i z a t i o n on tissue sections. We have applied the aforementioned methods to assess lineage of cytogenetically aberrant cells in biphaeic epithelialmesenchymal solid tumors. Clonal chromosome aberrations were demonstrated in mesenchymal components of breast adenofibromas, endometrial polyps, and pulmonary hamartomas. Epithelial cells, by contrast, were uniformly diploid. These studies indicate a mesenchymal origin in several biphasic solid tumors.
IIITIU..- U I O I S T I ~ - G ~
GI~ETIC
HTBRIDZS/tTIOW IS P ~ I C AD~zseNA MA Micale, JS Sanford, WA Sakr, IJ Poweli, and SR Wollan, DepartJenta of Pathology and 0roloc]y, Harper Hospital and Wayne State University School of Medicine, Detroit, MI. The application of molecular cytogenetic analysis to paraffin-blocked tissue sections utilizing fluorescence in situ hybridization (FISH) per~tits resolution of iaauea that have plagued conventional cytogenetic analysis of prostatic adenocarcinoHNLa. P r o b l e ~ including uncertainty of origin of cells analyzed, selection of normal diploid cells in culture, and hiatologic grade heterogeneity interfere with or reduce the frequency of detection of genetic alterations. Paraffin sections of prostatic tt~ora from thirtsen patlenta with advanced disease were analyzed by FISH utilizing paracentro1~ric DNA probes for chromoaomsa 4, 7, 8, I0, and the ¥ chrommeome. Kicronodular loss or gain was observed for all chromosomes except for ohrow~ao~e 4 which was used as the norm for expectation of diooaic frequency (it has not been associated with cytogenetic or molecular alteration in prostate cancer). Foci which demonstrated m/cronodular loaa or gain with one c h r o m o a ~ were often mutually exclusive of other loci in the a4u~e section that showed loss or galn of another chr~m~. The ga£nm o b s e r v e d in tumor for chromoeomms 7, 8, and the Y chromosome were not seen in aurrotu~itng h~perplaat/o tlaauss. Significant nodular loss of the Y cbromoaoee was Observed in two patienta end of chre~w~aome I0 in one patient. In other caaes, adjacent glands within tumor revealed gradual differenoea in galn or loss, suggesting another type of genetic non-unlformlty in prostate tumors. Probl4unm in interpretatlon related to aaaeaa~nt of non-intact and overlappiug nuclei, and to variation in hybridization efficiency among tumors and among different probes applied to the same tumor. The extent of heterogeneity demonstrated in these prostatic cancers may reflect advanced disease but is difficult to reconcile wlth the specificity of chromoa~ alteration assumed to be important in tumorigenesis.
26
FLUORESCENCE IN SITU H Y g R I D ~ A T I O N (FISH) ANALYSISOF PANCREATIC A D E N O C A R C I N O M A C. Griffin, P. Long, ILIL Hruban, C_ Yeo.
Ontology Center a n d Depts. of Pathology a n d Surgery,
The Johns Hopkins School Of Medicine, Balamore, MD Pancreatic adenocarcinomas (PA) often induce an intense non-neoplastic stromal cell reaction which may be the source of normal karyotypes obtained from some tumors. Since PA cells can be distinguished from scromal cells in intact tissue sections(TS), FISH provides a unique tool for identification of c h r o m o s o m a l abnormalities in neoplastic cells. We compared the results from centromere probes hybridized to interphase cells from primary tumors prepared directly for chromosome analysis (DP) to analysis of "IS from the same specimens. FISH analysis detected Irisomy 11 in 1/5 DPs vs 4 / 5 f o u n d in classical cytogenetic preparations of the same specimens. Trtsomy 12 was detected in 0 / 5 DPs while significant precentages of trisomy (20-44%) a n d t e r r a . m y (4055%) were found in 3/3 TS when only neoplastic cells were analyzed. Less than 1096 of cells h a d >2 copies of chromosome 20 in 5 / 5 DIs vs 25-34% of cells in 2 / 2 "IS. These data emphasize the significant contamination of P A with stromal cellsa n d the need for caution in the
interpretation of classical cytogenetics, FISH of cytogenetic preparations, a n d molecular analyses of these tumors, in situ adenocarcinomas USA) are thought to be precursor lesions in the development of infiltrating PA. Therefore, genetic analysis of these in s i s lesions might provide insight into the early events in development of PA. We used FISH with centromere probes to study ISA lesions. 0 / 8 lesions h a d >3 signals with chr 10, 1/20 lesions h a d >3 signals with chr 11, a n d 9 / 2 4 lesions had >3 signals with c h r 12, suggesting that there are some detectable trisomies in these early lesions. Additional characterization is underway.
THE RECURRENT CHROMOSOME CHANGES IN 29 SQUAMOUS CELL CARCINOMAS OF THE HEAD AND NECK REGION. Daniel L. Van Dyke, Maria J. Worsham, Michael J. Benninger, Thomas E. Carey. Henry Ford Hospital, D e t r o i t , MI; Univ o f Michigan, Ann Arbor. MI Our group has characterized the breakpoints, gains, and losses of chromosome material in squamous cell carcinoma (SCC) of the head and neck region from 29 patients. Cell lines were studied in I/3 of cases, direct preparations or early in vitro harvests in 1/3, and both in 1/3 of cases. GTG-banding was employed in all cases, as were C-banding and RBG- and AgNORstaining in most cases. Some tumors were near diploid and some near tetraploid, but many had mixed populations with diploid, tetraploid, and octoploid subclones. The most frequent changes were deletions: 3p13-p14, 8p21-p23, or IBqZl-q23 loss was observed in 28
~60Z of tumors; l o s s of the i n a c t i v e Xp in 71l of tumors from females; l o s s of Y in 63I from males; and loss of 4p12-p16, 21q. 9p, and 10p occurred in >40I of tumors. Including 6 patients with 5CC of the vulva, loss of 18q was observed in 14/16 patients who died with disease, and 2/3 alive with disease, but in only 1/12 who are alive with no evidence of disease. There was gain (usually 2 - 5z) of lq25-qter, 5p, 7p, and llql3-qter in 35-45I of tumors. Two tumors had an hsr at 11q13 which may be associated with amplification of the PRADI cyclin gene at that locus. A common breakpoint region in 9p (60t of tumors) appeared to be associated with loss of 9p, and icen-lp22 breakpoints, in 701, with loss of ip or duplication lq. An llp15 breakpoint, in 35I, correlated with loss of expression of cell surface antigen defined by monoclonal antibody UM-E7. An unbalanced t(l;acrocentric or 9)(pll;pll) in 30Z was usually associated with loss of ip. The emerging karyotypic pattern reflects a constellation of genetic changes in the evolution of SCC and presents a challenge to those who wish to characterize the earliest and the clinically important changes.
147
LINEAGE-REBTRICTED CKROMOSOME U E R R A TIOH8 INDICATE CELL OF ORIGIN IN BIPHA8IC 8SLID TOMOR8. Fletcher JA. Departments of Pathology and Pediatrics, H a r v a r d M e d i c a l S c h o o l , B o s t o n , MA. 25
Many solid tumors contain a single welldefined neoplastic cell population. Others contain two or more histologically distinct cell lineages that might potentially be neoplastic. Several approaches can be used to determine the cell lineages responsible for clone1 chromosome aberrations in solid tumors. These approaches include: 1) selective tissue culture of different cell populations prior to metaphase cell h a r v e s t s ; 2) i m m u n o h i s t o c h e m i c a l staining of m e t a p h a s e cells; 3) i m m u n o h i s t o c h e m i c a l staining coupled with chromosomal in situ hybridization in interphase cells; a n d 4) chromosomal in s i t u h y b r i d i z a t i o n on tissue sections. We have applied the aforementioned methods to assess lineage of cytogenetically aberrant cells in biphaeic epithelialmesenchymal solid tumors. Clonal chromosome aberrations were demonstrated in mesenchymal components of breast adenofibromas, endometrial polyps, and pulmonary hamartomas. Epithelial cells, by contrast, were uniformly diploid. These studies indicate a mesenchymal origin in several biphasic solid tumors.
IIITIU..- U I O I S T I ~ - G ~
GI~ETIC
HTBRIDZS/tTIOW IS P ~ I C AD~zseNA MA Micale, JS Sanford, WA Sakr, IJ Poweli, and SR Wollan, DepartJenta of Pathology and 0roloc]y, Harper Hospital and Wayne State University School of Medicine, Detroit, MI. The application of molecular cytogenetic analysis to paraffin-blocked tissue sections utilizing fluorescence in situ hybridization (FISH) per~tits resolution of iaauea that have plagued conventional cytogenetic analysis of prostatic adenocarcinoHNLa. P r o b l e ~ including uncertainty of origin of cells analyzed, selection of normal diploid cells in culture, and hiatologic grade heterogeneity interfere with or reduce the frequency of detection of genetic alterations. Paraffin sections of prostatic tt~ora from thirtsen patlenta with advanced disease were analyzed by FISH utilizing paracentro1~ric DNA probes for chromoaomsa 4, 7, 8, I0, and the ¥ chrommeome. Kicronodular loss or gain was observed for all chromosomes except for ohrow~ao~e 4 which was used as the norm for expectation of diooaic frequency (it has not been associated with cytogenetic or molecular alteration in prostate cancer). Foci which demonstrated m/cronodular loaa or gain with one c h r o m o a ~ were often mutually exclusive of other loci in the a4u~e section that showed loss or galn of another chr~m~. The ga£nm o b s e r v e d in tumor for chromoeomms 7, 8, and the Y chromosome were not seen in aurrotu~itng h~perplaat/o tlaauss. Significant nodular loss of the Y cbromoaoee was Observed in two patienta end of chre~w~aome I0 in one patient. In other caaes, adjacent glands within tumor revealed gradual differenoea in galn or loss, suggesting another type of genetic non-unlformlty in prostate tumors. Probl4unm in interpretatlon related to aaaeaa~nt of non-intact and overlappiug nuclei, and to variation in hybridization efficiency among tumors and among different probes applied to the same tumor. The extent of heterogeneity demonstrated in these prostatic cancers may reflect advanced disease but is difficult to reconcile wlth the specificity of chromoa~ alteration assumed to be important in tumorigenesis.
26
FLUORESCENCE IN SITU H Y g R I D ~ A T I O N (FISH) ANALYSISOF PANCREATIC A D E N O C A R C I N O M A C. Griffin, P. Long, ILIL Hruban, C_ Yeo.
Ontology Center a n d Depts. of Pathology a n d Surgery,
The Johns Hopkins School Of Medicine, Balamore, MD Pancreatic adenocarcinomas (PA) often induce an intense non-neoplastic stromal cell reaction which may be the source of normal karyotypes obtained from some tumors. Since PA cells can be distinguished from scromal cells in intact tissue sections(TS), FISH provides a unique tool for identification of c h r o m o s o m a l abnormalities in neoplastic cells. We compared the results from centromere probes hybridized to interphase cells from primary tumors prepared directly for chromosome analysis (DP) to analysis of "IS from the same specimens. FISH analysis detected Irisomy 11 in 1/5 DPs vs 4 / 5 f o u n d in classical cytogenetic preparations of the same specimens. Trtsomy 12 was detected in 0 / 5 DPs while significant precentages of trisomy (20-44%) a n d t e r r a . m y (4055%) were found in 3/3 TS when only neoplastic cells were analyzed. Less than 1096 of cells h a d >2 copies of chromosome 20 in 5 / 5 DIs vs 25-34% of cells in 2 / 2 "IS. These data emphasize the significant contamination of P A with stromal cellsa n d the need for caution in the
interpretation of classical cytogenetics, FISH of cytogenetic preparations, a n d molecular analyses of these tumors, in situ adenocarcinomas USA) are thought to be precursor lesions in the development of infiltrating PA. Therefore, genetic analysis of these in s i s lesions might provide insight into the early events in development of PA. We used FISH with centromere probes to study ISA lesions. 0 / 8 lesions h a d >3 signals with chr 10, 1/20 lesions h a d >3 signals with chr 11, a n d 9 / 2 4 lesions had >3 signals with c h r 12, suggesting that there are some detectable trisomies in these early lesions. Additional characterization is underway.
THE RECURRENT CHROMOSOME CHANGES IN 29 SQUAMOUS CELL CARCINOMAS OF THE HEAD AND NECK REGION. Daniel L. Van Dyke, Maria J. Worsham, Michael J. Benninger, Thomas E. Carey. Henry Ford Hospital, D e t r o i t , MI; Univ o f Michigan, Ann Arbor. MI Our group has characterized the breakpoints, gains, and losses of chromosome material in squamous cell carcinoma (SCC) of the head and neck region from 29 patients. Cell lines were studied in I/3 of cases, direct preparations or early in vitro harvests in 1/3, and both in 1/3 of cases. GTG-banding was employed in all cases, as were C-banding and RBG- and AgNORstaining in most cases. Some tumors were near diploid and some near tetraploid, but many had mixed populations with diploid, tetraploid, and octoploid subclones. The most frequent changes were deletions: 3p13-p14, 8p21-p23, or IBqZl-q23 loss was observed in 28
~60Z of tumors; l o s s of the i n a c t i v e Xp in 71l of tumors from females; l o s s of Y in 63I from males; and loss of 4p12-p16, 21q. 9p, and 10p occurred in >40I of tumors. Including 6 patients with 5CC of the vulva, loss of 18q was observed in 14/16 patients who died with disease, and 2/3 alive with disease, but in only 1/12 who are alive with no evidence of disease. There was gain (usually 2 - 5z) of lq25-qter, 5p, 7p, and llql3-qter in 35-45I of tumors. Two tumors had an hsr at 11q13 which may be associated with amplification of the PRADI cyclin gene at that locus. A common breakpoint region in 9p (60t of tumors) appeared to be associated with loss of 9p, and icen-lp22 breakpoints, in 701, with loss of ip or duplication lq. An llp15 breakpoint, in 35I, correlated with loss of expression of cell surface antigen defined by monoclonal antibody UM-E7. An unbalanced t(l;acrocentric or 9)(pll;pll) in 30Z was usually associated with loss of ip. The emerging karyotypic pattern reflects a constellation of genetic changes in the evolution of SCC and presents a challenge to those who wish to characterize the earliest and the clinically important changes.