Chromosome Abnormalities in Lymphocytes and Fibroblasts of Subjects with Multiple Endocrine Neoplasia Type 1 Susi Scappaticci, Paola Maraschio, Nicola del Ciotto, Gian Si|vio Fossati, Aris Zonta, and Marco Fraccaro
ABSTRACT: A consistent degree of chromosome instability was found in cultured lymphocytes and in fibroblasts derived from skin biopsies of three patients with multiple endocrine neoplasia type 1 (MEN1). In both tissues, there was a significant increase in chromasamal breakage. Dicentrics and tricentries, rings, translacations, deletions, acentrie fragments, and presumptive double minutes were the most fl'equent abnormalities in lymphocytes. The fibrablusts of two patients had large clones consisting of trisamy 7 and trisumy 18 cells, respectively.
INTRODUCTION
Multiple endocrine neoplasia type 1 (MEN1) is an antosomal d o m i n a n t condition that predisposes to neoplasia of the parathyroid glands and to hyperplasia or tumors of the anterior pituitary and the endocrine pancreas. Patients may also develop foregut carcinoid tumors and lipomas [1]. Linkage analysis with the gene for skeletal muscle glycogen phosphorylase (PYGM) has localized the gene for MEN1 to the long arm of chromosome 11 within band q13 telomeric to the PYGM locus [2, 3]. Cytogenetic investigations limited to the scoring of chromosome breakage in Giemsa-stained preparations of cultured lymphocytes showed no consistent pattern of chromosomal instability in family members with MEN1, but several individuals in the affected families clearly had an increased n u m b e r of chromosome breakage as compared with healthy controls I4]. We report three patients with MEN1 who had chromosomal abnormalities in lymphocytes and in fibroblasts cultured from biopsies of healthy skin. CASE R E P O R T S
Case 1
A 43-year-old man had n u m e r o u s cutaneous fibromas distributed laterally along the thoracoabdominal nerves. At 41, he had a hemorrhagic shock due to a duodenal ulcer. One year later, his thyroid and parathyroids were surgically removed, owing to the From BiologiaGenerale e GeneticaMedica (S. S., P, M., M. F,), and Dipartimentodi Chirurgia(N. d. (;., G. S. F., A. Z.], Universitadi Pavia, Italy. Address reprint requests to: Marc() Fraccaro, C. P., 217. 27100 Pavia, Italy. Received April 27, 1990; accepted July 11, 1990.
85 ('i 1991 ElsevierSciencePublishingCo., lnc. 655 Awmtmof the Americas, New York, NY 10010
(:an(:er(;enetCytogem!t52:85 92 {19911 01115-46(18/91/$03.511
86 Table 1
D e t a i l e d a n a l y s i s of m e t a p h a s e s w i t h c h r o m o s o m e a b n o r m a l i t y a n d / o r aneuploidy in lymphocyte cultures No. of cells analyzed
No. of cells with breaks
Abnormal cells
1
150
12
36
2
120
8
17
Case
Detailed analysis of abnormal cells 1) 43,XY, 3 . - 4 , 7.-22.+tri(3;4;7)(3pter~. 3qter::4pter-~4qter::7pter~ 7qter] 2) 44.XY, 5 , - 1 0 , 1 6 , - 2 0 . + d i c ( 5 ; 1 0 ) (5pter--~5ql 3::10q22--,10pter) + die(16;20/ (16pter--. 16qter::20pter---*20qter/ 3) 44.XY, 2 . - 1 3 , 1 4 , - 1 8 , - 2 2 , + d e 1 ( 1 4 ) (14pter--~l 4q11), + t(13;22)(13qter~ 13q22::22ql 3--~22pter), + dic(13;14;2) {13pter--~ 13q22:14ql 1--. 14qter:2pter--~2qter). 4) 45,XY,-5, 10,+dic(5;10)(5pter--~ 5ql 3::10q22--~10pter) 5] 44,XY, 1 , - 6 , 16,+dic(1;6)(q?,p?),+ace 6) 44.XY, 4 , - 6 , 7,+dic(4;7)(q21;q36),+ace 7) 45,XY,- 1 9 , - 22.t(13;19)(p11;q22), + dic(19;22) (19pter--.19qter:22qter--~22 pter) 8) 43.XY,- 10,- 21,-22,del(1)(q12) 9) 46,XY.del(2)(q11), + ace 10) 46,XY,t(2 ;4)(qll ;q21) 11) 47,XY, + r(18) 12) 47,XY, 2, + r(?). t mar 13) 47,XY. 2,+r(?).+mar(PCC) 14) 46,XY,inv(2)(p13q37) 15) 46,XY(PCC) 16) 45.XY,+3. 4, 7 17, 18) 4 5 . X Y . - 1 7 , + r a i n 19) 45,XY,- 20,~ rain 20-22) 46,XY, t rain 23) 45.XY,- 1 7 , + d r a i n 24-32) 46,XY. + drain 33-34) 46.XY, + dmin 35) 45,XY, 8 36) 45.XY, 21 1) 45,XX, - 21,t(18;21)(q11;pl 1) 2) 46,XX,t(18;22)(q22;q11) 3) 45.XX.- 18,r(1) 4) 46,X.PCD(X),+PCD(X), 9 5) 45.XX,- 12,+ ace 6) 46.XX. + 2ace 7, 8) 46,XX, + rain 9, 10) 46,XX,+ drain 11) 46,X,PCD(X). + dmin 12) 45,XX,- 17 13. 14) 45,X, X 15) 4 6 , X , - X . + 17 16, 17) 47.XX.+X (continued)
C h r o m o s o m e Abnormalities in Multiple Endocrine Neoplasia
Table 1
87
Continued No. {}f
No. of
(:ells analyzed
cells with breaks
3
150
13
('ontrols/ Sex/Age (yr) 1/F/4(I 2/M/50 3/F/56 4/M/40 5/F/60
100 100 100 100 100
2 3 4 4 3
Case
Abnormal ceils 9
l}elailed analysis of abnormal cells 1)44,XX, 16, 17, 20,+1ri(16;20;17) (16pter~ 16qtcr::20pter 20qter::l 7qter~ 171)ter) 2} 46,XX,r(7) 3) 48,XX, + r(?),+ r[?) 4) 46,XX,{:{mslri{:tiol~ 12(q22] 5} 45.XX. 13 6} 45,XX, 19 7, 8) 45,XX, 17 9) >,qO,XXXX,+ PCC,+ dmin
Abbreviations: P(X2 I}reillaltlr{~ chronloS(Hll{} {:olld{~lisa[iOll; P(~D. ]lr[~mattlr(~ c(}ntrolnere divisioll,
presence of adenomas. The patient now has a pancreatic adenoma. The father {now dead) of the patient was also affected. Case 2
A 49-year-old w o m a n had duodenal ulcers at age 34. Parathyroidectomy was performed w h e n she was aged 44 years to remove an adenoma. At age 45, the patient u n d e r w e n t total gastrectomy and distal subtotal resection of the pancreas to remove an adenoma. The father and one brother {dead) were also affected. Case 3
A 64-year-old w o m a n had renal stones at age 42 and a duodenal ulcer at age 52. She u n d e r w e n t subtotal gastrectocny for central and duodenal ulcer. At age 63, she u n d e r w e n t parathyroidectomy to remove an adenoma. There are no other known affected members in the family. None of the patients u n d e r w e n t tumor chemotherapy or radiation treatment.
MATERIALS
AND METHODS
Peripheral blood samples stimulated by phytohemagglutinin cultured for 72 hours in RPMI 1640 m e d i u m s u p p l e m e n t e d with 15% fetal calf serum. Fibroblast cultures were derived from upper-arm skin biopsies and maintained in Ham's E l 0 m e d i u m s u p p l e m e n t e d with antiobiotics and 20% fetal bovine serum. Chromosome preparations were made after trypsinization at the first passage. As controls, we used lymphocytes and fibroblast cultures from five healthy subjects. Chromosome preparations were stained with Giemsa and for QFQ bands. When required, slides were stained for C-bands and DA-DAPI as well. We first scored more than 100 metaphases in each of the three patients and recorded
88
S. Scal)pati(:(:i et al.
a
~v
b
/' C
\
e
F i g u r e 1 Tri(:entric chromosoml!s with one, ina(:tive, cuntromere iI1 l vmphocytes of case I (a) and c:ase, 3 (b) Gie,msa. I)ic:entric ((:) and ring chromosomes in lymphocytes of (:ase 1 (c:} and 3 (d] Gielnsa. Double minutes in lymphocyl{~,s of case 1. (;iemsa (el and C-bands {f). all n u m e r i c a l a n d s t r u c t u r a l a b n o r m a l i t i e s . S u b s e q u e n t l y . we s c o r e d t h e cells w i t h c h r o m o s o i n e a n d c h r o m a t i d b r e a k s in c u l t u r e d l y m p b o c y t e s .
RESULTS Lymphocytes T h e f i n d i n g s in l y m p h o c y t e s are s u m m a r i z e d in T a b l e 1. T h e l y m p h o c y t e s s h o w e d 8(}{,. 7
ABSTRACT: A consistent degree of chromosome instability was found in cultured lymphocytes and in fibroblasts derived from skin biopsies of three patients with multiple endocrine neoplasia type 1 (MEN1). In both tissues, there was a significant increase in chromasamal breakage. Dicentrics and tricentries, rings, translacations, deletions, acentrie fragments, and presumptive double minutes were the most fl'equent abnormalities in lymphocytes. The fibrablusts of two patients had large clones consisting of trisamy 7 and trisumy 18 cells, respectively.
INTRODUCTION
Multiple endocrine neoplasia type 1 (MEN1) is an antosomal d o m i n a n t condition that predisposes to neoplasia of the parathyroid glands and to hyperplasia or tumors of the anterior pituitary and the endocrine pancreas. Patients may also develop foregut carcinoid tumors and lipomas [1]. Linkage analysis with the gene for skeletal muscle glycogen phosphorylase (PYGM) has localized the gene for MEN1 to the long arm of chromosome 11 within band q13 telomeric to the PYGM locus [2, 3]. Cytogenetic investigations limited to the scoring of chromosome breakage in Giemsa-stained preparations of cultured lymphocytes showed no consistent pattern of chromosomal instability in family members with MEN1, but several individuals in the affected families clearly had an increased n u m b e r of chromosome breakage as compared with healthy controls I4]. We report three patients with MEN1 who had chromosomal abnormalities in lymphocytes and in fibroblasts cultured from biopsies of healthy skin. CASE R E P O R T S
Case 1
A 43-year-old man had n u m e r o u s cutaneous fibromas distributed laterally along the thoracoabdominal nerves. At 41, he had a hemorrhagic shock due to a duodenal ulcer. One year later, his thyroid and parathyroids were surgically removed, owing to the From BiologiaGenerale e GeneticaMedica (S. S., P, M., M. F,), and Dipartimentodi Chirurgia(N. d. (;., G. S. F., A. Z.], Universitadi Pavia, Italy. Address reprint requests to: Marc() Fraccaro, C. P., 217. 27100 Pavia, Italy. Received April 27, 1990; accepted July 11, 1990.
85 ('i 1991 ElsevierSciencePublishingCo., lnc. 655 Awmtmof the Americas, New York, NY 10010
(:an(:er(;enetCytogem!t52:85 92 {19911 01115-46(18/91/$03.511
86 Table 1
D e t a i l e d a n a l y s i s of m e t a p h a s e s w i t h c h r o m o s o m e a b n o r m a l i t y a n d / o r aneuploidy in lymphocyte cultures No. of cells analyzed
No. of cells with breaks
Abnormal cells
1
150
12
36
2
120
8
17
Case
Detailed analysis of abnormal cells 1) 43,XY, 3 . - 4 , 7.-22.+tri(3;4;7)(3pter~. 3qter::4pter-~4qter::7pter~ 7qter] 2) 44.XY, 5 , - 1 0 , 1 6 , - 2 0 . + d i c ( 5 ; 1 0 ) (5pter--~5ql 3::10q22--,10pter) + die(16;20/ (16pter--. 16qter::20pter---*20qter/ 3) 44.XY, 2 . - 1 3 , 1 4 , - 1 8 , - 2 2 , + d e 1 ( 1 4 ) (14pter--~l 4q11), + t(13;22)(13qter~ 13q22::22ql 3--~22pter), + dic(13;14;2) {13pter--~ 13q22:14ql 1--. 14qter:2pter--~2qter). 4) 45,XY,-5, 10,+dic(5;10)(5pter--~ 5ql 3::10q22--~10pter) 5] 44,XY, 1 , - 6 , 16,+dic(1;6)(q?,p?),+ace 6) 44.XY, 4 , - 6 , 7,+dic(4;7)(q21;q36),+ace 7) 45,XY,- 1 9 , - 22.t(13;19)(p11;q22), + dic(19;22) (19pter--.19qter:22qter--~22 pter) 8) 43.XY,- 10,- 21,-22,del(1)(q12) 9) 46,XY.del(2)(q11), + ace 10) 46,XY,t(2 ;4)(qll ;q21) 11) 47,XY, + r(18) 12) 47,XY, 2, + r(?). t mar 13) 47,XY. 2,+r(?).+mar(PCC) 14) 46,XY,inv(2)(p13q37) 15) 46,XY(PCC) 16) 45.XY,+3. 4, 7 17, 18) 4 5 . X Y . - 1 7 , + r a i n 19) 45,XY,- 20,~ rain 20-22) 46,XY, t rain 23) 45.XY,- 1 7 , + d r a i n 24-32) 46,XY. + drain 33-34) 46.XY, + dmin 35) 45,XY, 8 36) 45.XY, 21 1) 45,XX, - 21,t(18;21)(q11;pl 1) 2) 46,XX,t(18;22)(q22;q11) 3) 45.XX.- 18,r(1) 4) 46,X.PCD(X),+PCD(X), 9 5) 45.XX,- 12,+ ace 6) 46.XX. + 2ace 7, 8) 46,XX, + rain 9, 10) 46,XX,+ drain 11) 46,X,PCD(X). + dmin 12) 45,XX,- 17 13. 14) 45,X, X 15) 4 6 , X , - X . + 17 16, 17) 47.XX.+X (continued)
C h r o m o s o m e Abnormalities in Multiple Endocrine Neoplasia
Table 1
87
Continued No. {}f
No. of
(:ells analyzed
cells with breaks
3
150
13
('ontrols/ Sex/Age (yr) 1/F/4(I 2/M/50 3/F/56 4/M/40 5/F/60
100 100 100 100 100
2 3 4 4 3
Case
Abnormal ceils 9
l}elailed analysis of abnormal cells 1)44,XX, 16, 17, 20,+1ri(16;20;17) (16pter~ 16qtcr::20pter 20qter::l 7qter~ 171)ter) 2} 46,XX,r(7) 3) 48,XX, + r(?),+ r[?) 4) 46,XX,{:{mslri{:tiol~ 12(q22] 5} 45.XX. 13 6} 45,XX, 19 7, 8) 45,XX, 17 9) >,qO,XXXX,+ PCC,+ dmin
Abbreviations: P(X2 I}reillaltlr{~ chronloS(Hll{} {:olld{~lisa[iOll; P(~D. ]lr[~mattlr(~ c(}ntrolnere divisioll,
presence of adenomas. The patient now has a pancreatic adenoma. The father {now dead) of the patient was also affected. Case 2
A 49-year-old w o m a n had duodenal ulcers at age 34. Parathyroidectomy was performed w h e n she was aged 44 years to remove an adenoma. At age 45, the patient u n d e r w e n t total gastrectomy and distal subtotal resection of the pancreas to remove an adenoma. The father and one brother {dead) were also affected. Case 3
A 64-year-old w o m a n had renal stones at age 42 and a duodenal ulcer at age 52. She u n d e r w e n t subtotal gastrectocny for central and duodenal ulcer. At age 63, she u n d e r w e n t parathyroidectomy to remove an adenoma. There are no other known affected members in the family. None of the patients u n d e r w e n t tumor chemotherapy or radiation treatment.
MATERIALS
AND METHODS
Peripheral blood samples stimulated by phytohemagglutinin cultured for 72 hours in RPMI 1640 m e d i u m s u p p l e m e n t e d with 15% fetal calf serum. Fibroblast cultures were derived from upper-arm skin biopsies and maintained in Ham's E l 0 m e d i u m s u p p l e m e n t e d with antiobiotics and 20% fetal bovine serum. Chromosome preparations were made after trypsinization at the first passage. As controls, we used lymphocytes and fibroblast cultures from five healthy subjects. Chromosome preparations were stained with Giemsa and for QFQ bands. When required, slides were stained for C-bands and DA-DAPI as well. We first scored more than 100 metaphases in each of the three patients and recorded
88
S. Scal)pati(:(:i et al.
a
~v
b
/' C
\
e
F i g u r e 1 Tri(:entric chromosoml!s with one, ina(:tive, cuntromere iI1 l vmphocytes of case I (a) and c:ase, 3 (b) Gie,msa. I)ic:entric ((:) and ring chromosomes in lymphocytes of (:ase 1 (c:} and 3 (d] Gielnsa. Double minutes in lymphocyl{~,s of case 1. (;iemsa (el and C-bands {f). all n u m e r i c a l a n d s t r u c t u r a l a b n o r m a l i t i e s . S u b s e q u e n t l y . we s c o r e d t h e cells w i t h c h r o m o s o i n e a n d c h r o m a t i d b r e a k s in c u l t u r e d l y m p b o c y t e s .
RESULTS Lymphocytes T h e f i n d i n g s in l y m p h o c y t e s are s u m m a r i z e d in T a b l e 1. T h e l y m p h o c y t e s s h o w e d 8(}{,. 7
