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Cytogenetic analysis of a multinodular thyroid goiter
Cytogenetic Analysis of a Multinodular Thyroid Goiter Nilce B. Gama, Reginaldo Gama, Jorge A. Thom , and Eloiza H. Tajara
ABSTRACT: Sh•rt•term cu•tures •f a c•••agenase
disaggregated mu•tin•du•ar g•iter was sh•wn by cyt•ge. netic analysis to have the mosaic karyotype 47,XX, + 7/48,XX, + 7, + 17/49,XX, + 7, + 10, + 17. No cytogenetic data on goiter are available for comparison with the present case.
INTRODUCTION Thyroid nodular hyperplasia or nontoxic nodular goiter is the most c o m m o n thyroid disease and may be defined as a thyroid enlargement due to repeated or c o n t i n u o u s hyperplasia, usually in response to thyroid h o r m o n e deficiency. The possibility that this type of lesion is associated with an increased carcinoma i n ci d en ce is still unresolved. If such increase in i n c id e n c e exists, it is probably slight [1]. We describe three related cytogenetic clones in a nontoxic m u l t i n o d u l a r goiter.
CASE REPORT A 43-year-old w o m a n had noticed enlargement of her thyroid for more than 30 years. During the m o n t h before she was hospitalized, she had referred local pain. A subtotal t h y ro i d ect o m y was performed; pathologic examination of the excised mass, w h i c h measured 70 × 50 × 25 mm, showed a m u l t i n o d u l a r goiter with excessive accumulation of colloid and over-sized follicles (Fig. 1).
CYTOGENETIC STUDIES The freshly obtained s p e c i m e n was processed for short-term cultures as previously described [2]. The sample was finely m i n c e d with scissors, disaggregated in a collagenase solution, and plated in flasks in Ham's F-10 m e d i u m s u p p l e m e n t e d with fetal calf serum, glutamine, antibiotics, and insulin. All cultures were harvested w i t h i n 7 days by Colcemid exposure followed by hypotonic treatment and methanol : acetic fixation (3 : 1). G banding was obtained by trypsin and Giemsa stain.
From Departamento de Biologia, Instituto de Bioci~ncias, Letras e Ci~ncias Exatas de S~o Jos~ do Rio Preto, UNESP(N. B. G., E, H. T.); Departamentos de Cirurgia (R. G.) e Patologia (J. A. T.), Faculdade Regional de Medicina, S~o Jos~ do Rio Preto, SP, Brazil. Address reprint requests to: Dr. E. H. Tajara, Departamento de Biologia, IBILCE, UNESP, Caixa Postal 136, 15055 S~o Jos~ do Rio Preto, SP, Brazil. Received September 4, 1990; accepted January 10, 1991.
73 © 1991 Elsevier Science Publishing Co., Inc. 655 Avenue of the Americas. New York, NY 10010
Cancer Genet Cytogenet 55:73-77 (1991) 0165-4608/91/$03.50
F i g u r e I Histologic section of the multinodular goiter showing oversized follicles and exce sive accumulation of colloid (H&E × 160).
Figure 2
Representative karyotype of the clone 47,XX, + 7.
B 1
2
6
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13
7
3
4
8
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E
14
20
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JJ 22
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Cytogenetics of M u l t i n o d u l a r Thyroid Goiter
75
1"t ...... .e.t':...i!... B l
2
4
3
5
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7
......
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9
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Figure 3
20
21
22
12
~::"' ~ , ' ~ 17
•i i
....
|~ 18
........... ×
Y
Representative karyotype of the clone 48,XX, + 7, + 17.
Eighty metaphases were partially or completely analyzed. In 11 metaphases the presumed stemline karyotype 47,XX, + 7 was observed. Two cells had the stemline pattern with additional trisomy 17, and n i n e cells showed trisomies 7 and 17 as well as trisomy 10. Each population was detected in at least two culture flasks. No other trisomies were observed except in one cell with trisomies 2 and 16 and some monosomies. Two polyploid cells and r a n d o m chromosome losses in apparently normal cells or from abnormal clones were also observed. Representative karyotypes from the abnormal clones are shown in Figures 2-4.
DISCUSSION
The chromosome pattern of the cell population from this m u l t i n o d u l a r goiter is compatible with clonal evolution from a cell that u n d e r w e n t n o n d i s j u n c t i o n of chromosome 7 and subsequent n o n d i s j u n c t i o n s of chromosomes 17 and 10. Besides the normal cells, the clone 47,XX, + 7 was the most frequent in the goiter cell population. To our knowledge, there are only a few previous reports on cytogenetic studies of thyroid lesions and none on goiter. Therefore, no data are available for comparison with our case. The observations of 62 thyroid carcinomas and adenomas have shown abnormalities of chromosome 10 in ten tumors, polysomy 7 in two, and structural and numerical alterations involving other chromosomes in 12 of them [3-7].
76
N.B. Gama et al.
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Figure 4
1
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12
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E
14
i ~ ........ 19
.
5
J 20
16
15
A G-
A .......
~41
21
22
17
18
1-IX .......... Y
Representative karyotype of the clone 49,XX, + 7, + 10, + 17.
Trisomies 7, 10, and 17 are common findings in hematologic malignancies and in solid tumors [8]. In addition, different genes located on chromosome 7, 10, and 17 are related to cell proliferation or thyroid function such as the epidermal growth factor receptor (EGFR) [9], the transforming sequence, thyroid i (TST1), the thyroid stimulating hormone receptor-like 3 [10], and the protooncogenes c-erb A homologs encoding proteins with different T3 binding properties [11]. Thus, trisomy 7 associated with trisomies 10 and 17 detected in the goiter may be connected with the mechanism of the disease through genes related to cell proliferation or thyroid function. However, trisomies 7 and 10 have been reported in short-term cultures of nonneoplastic cells from patients with cancer [12, 13]; these findings may represent genomic instability in the tumor parenchyma or may characterize stromal fibroblasts [14].
This study was supported by Conselho Nacional de Desenvolvimento Cientifico e Tecnol6gico (CNPq), Funda§~o para o Desenvolvimento da UNESP (FUNDUNESP), Funda§8o de Amparo Pesqnisa do Estado de $8o Paulo (FAPESP) and Coordena~8o de Aperfei~oamento do Pessoal de Ensino Superior (CAPES). The authors thank Josu6 Rodrigues dos Santos and Rosana Silistino de Souza for technical assistance, Dr. Marileila Varella-Garcia for helpful discussion, and Dr. Carlos Daghlian for reviewing the text.
C y t o g e n e t i c s of M u l t i n o d u l a r T h y r o i d G o i t e r
77
REFERENCES 1. Rosai J (1989): Ackerman's Surgical Pathology, Vol. 1, 7th ed., C.V. Mosby, St. Louis, pp. 391-447. 2. Limon J, Dal Cin P, Sandberg AA (1986): Application of long term collagenase disaggregation for the cytogenetic analysis of h u m a n solid tumors. Cancer Genet Cytogenet 23:305317. 3. Wurster-Hill DH, Noll WW, Bircher LY, Pettengill OS, Grizzle WA (1986): Cytogenetics of medullary carcinoma of the thyroid. Cancer Genet Cytogenet 20:247-253. 4. Antonini P, Venuat AM, Linares G, Caillou B, Berger R, Parmentier C (1989): A translocation (7;10)(q35;q21) in a differentiated papillary carcinoma of the thyroid. Cancer Genet Cytogenet 41:139-144. 5. Bondeson L, Bengtsson A, Bondeson AG, Dahlenfors R, Grimelius L, Wedell B, Mark J (1989): Chromosome studies in thyroid neoplasia. Cancer 64:680-685. 6. Herath J, Jenkins R, Hay I, Grant C, Dewald G (1989): Cytogenetic studies on 12 thyroid carcinomas [Abstract]. In: International Workshop on Chromosomes in Solid Tumors, 3, Tucson, AZ, February 26 to March 3, 1989. Tucson, The Arizona Cancer Center and The National Cancer Institute, p. 64. 7. Herrmann ME, Bullerdiek J, Bartnitzke S, Lebert B, Lobeck H, Zuschneid W (1989): Cytogenetic studies on 15 thyroid adenomas [Abstract]. In: International Workshop on Chromosomes in Solid Tumors, 3, Tucson, AZ, February 26 to March 3, 1989. Tucson, The Arizona Cancer Center and The National Cancer Institute, p. 65. 8. Mitelman F (1985): Catalog of Chromosome Aberrations in Cancer, Vol. 5, 2nd ed. Alan R. Liss, New York, pp. 1-707. 9. Davies RL, Grosse VA, Kucherlapati R, Bothwell M (1980): Genetical analysis of epidermal growth factor action: Assignment of h u m a n epidermal growth factor receptor gene to chromosome 7. Proc Natl Acad Sci USA 77:4188-4192. 10. Smith M, Simpson NE (1989): Report of the committee on the genetic constitution of chromosomes 9 and 10. Cytogenet Cell Genet 51:202-225. 11. Miyajima N, Horiuchi R, Shibuya Y, Fukushige S, Matsubara K, Toyoshima K, Yamamoto T (1989): Two erbA homologs encoding proteins with different T3 binding capacities are transcribed from opposite DNA strands of the same genetic locus. Cell 57:31-39. 12. Heim S, Lindstrom E, Salford, LG (1990): Trisomy 7 and sex chromosome loss are not tumor-specific aberrations in malignant gliomas. Abstracts of 2nd European Workshop on Cytogenetics and Molecular Genetics of Human Solid Tumours: A18. 13. Szucs S, Emanuel A, Kovacs G (1990): Clonal aberrations of chromosomes X, Y, 7, and 10 in normal kidney tissue of patients with renal cell cancers. Abstracts of 2nd European Workshop on Cytogenetics and Molecular Genetics of Human Solid Tumours: P40. 14. Bardi G, Johansson B, Orndal C, Heim S, Mandahl N, Pandis N, Andren-Sandberg A, Mitelman F (1990): Abstracts of 2nd European Workshop on Cytogenetics and Molecular Genetics of Human Solid Tumours: P22.
ABSTRACT: Sh•rt•term cu•tures •f a c•••agenase
disaggregated mu•tin•du•ar g•iter was sh•wn by cyt•ge. netic analysis to have the mosaic karyotype 47,XX, + 7/48,XX, + 7, + 17/49,XX, + 7, + 10, + 17. No cytogenetic data on goiter are available for comparison with the present case.
INTRODUCTION Thyroid nodular hyperplasia or nontoxic nodular goiter is the most c o m m o n thyroid disease and may be defined as a thyroid enlargement due to repeated or c o n t i n u o u s hyperplasia, usually in response to thyroid h o r m o n e deficiency. The possibility that this type of lesion is associated with an increased carcinoma i n ci d en ce is still unresolved. If such increase in i n c id e n c e exists, it is probably slight [1]. We describe three related cytogenetic clones in a nontoxic m u l t i n o d u l a r goiter.
CASE REPORT A 43-year-old w o m a n had noticed enlargement of her thyroid for more than 30 years. During the m o n t h before she was hospitalized, she had referred local pain. A subtotal t h y ro i d ect o m y was performed; pathologic examination of the excised mass, w h i c h measured 70 × 50 × 25 mm, showed a m u l t i n o d u l a r goiter with excessive accumulation of colloid and over-sized follicles (Fig. 1).
CYTOGENETIC STUDIES The freshly obtained s p e c i m e n was processed for short-term cultures as previously described [2]. The sample was finely m i n c e d with scissors, disaggregated in a collagenase solution, and plated in flasks in Ham's F-10 m e d i u m s u p p l e m e n t e d with fetal calf serum, glutamine, antibiotics, and insulin. All cultures were harvested w i t h i n 7 days by Colcemid exposure followed by hypotonic treatment and methanol : acetic fixation (3 : 1). G banding was obtained by trypsin and Giemsa stain.
From Departamento de Biologia, Instituto de Bioci~ncias, Letras e Ci~ncias Exatas de S~o Jos~ do Rio Preto, UNESP(N. B. G., E, H. T.); Departamentos de Cirurgia (R. G.) e Patologia (J. A. T.), Faculdade Regional de Medicina, S~o Jos~ do Rio Preto, SP, Brazil. Address reprint requests to: Dr. E. H. Tajara, Departamento de Biologia, IBILCE, UNESP, Caixa Postal 136, 15055 S~o Jos~ do Rio Preto, SP, Brazil. Received September 4, 1990; accepted January 10, 1991.
73 © 1991 Elsevier Science Publishing Co., Inc. 655 Avenue of the Americas. New York, NY 10010
Cancer Genet Cytogenet 55:73-77 (1991) 0165-4608/91/$03.50
F i g u r e I Histologic section of the multinodular goiter showing oversized follicles and exce sive accumulation of colloid (H&E × 160).
Figure 2
Representative karyotype of the clone 47,XX, + 7.
B 1
2
6
t
13
7
3
4
8
.
.
.
E
14
20
-
15
21
JJ 22
5
II
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~I
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iii
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,__4 19
I0
9
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tl
17
18
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.
Y
Cytogenetics of M u l t i n o d u l a r Thyroid Goiter
75
1"t ...... .e.t':...i!... B l
2
4
3
5
...... •i i t l 6
7
......
13
~'e
..... It
ilJ
8
lO
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........
14
.......
9
I i
15
•
II
.......
16
" ~ ' 8
.......
~'"
G 19
Figure 3
20
21
22
12
~::"' ~ , ' ~ 17
•i i
....
|~ 18
........... ×
Y
Representative karyotype of the clone 48,XX, + 7, + 17.
Eighty metaphases were partially or completely analyzed. In 11 metaphases the presumed stemline karyotype 47,XX, + 7 was observed. Two cells had the stemline pattern with additional trisomy 17, and n i n e cells showed trisomies 7 and 17 as well as trisomy 10. Each population was detected in at least two culture flasks. No other trisomies were observed except in one cell with trisomies 2 and 16 and some monosomies. Two polyploid cells and r a n d o m chromosome losses in apparently normal cells or from abnormal clones were also observed. Representative karyotypes from the abnormal clones are shown in Figures 2-4.
DISCUSSION
The chromosome pattern of the cell population from this m u l t i n o d u l a r goiter is compatible with clonal evolution from a cell that u n d e r w e n t n o n d i s j u n c t i o n of chromosome 7 and subsequent n o n d i s j u n c t i o n s of chromosomes 17 and 10. Besides the normal cells, the clone 47,XX, + 7 was the most frequent in the goiter cell population. To our knowledge, there are only a few previous reports on cytogenetic studies of thyroid lesions and none on goiter. Therefore, no data are available for comparison with our case. The observations of 62 thyroid carcinomas and adenomas have shown abnormalities of chromosome 10 in ten tumors, polysomy 7 in two, and structural and numerical alterations involving other chromosomes in 12 of them [3-7].
76
N.B. Gama et al.
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iii
2
1
6
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Figure 4
1
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12
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E
14
i ~ ........ 19
.
5
J 20
16
15
A G-
A .......
~41
21
22
17
18
1-IX .......... Y
Representative karyotype of the clone 49,XX, + 7, + 10, + 17.
Trisomies 7, 10, and 17 are common findings in hematologic malignancies and in solid tumors [8]. In addition, different genes located on chromosome 7, 10, and 17 are related to cell proliferation or thyroid function such as the epidermal growth factor receptor (EGFR) [9], the transforming sequence, thyroid i (TST1), the thyroid stimulating hormone receptor-like 3 [10], and the protooncogenes c-erb A homologs encoding proteins with different T3 binding properties [11]. Thus, trisomy 7 associated with trisomies 10 and 17 detected in the goiter may be connected with the mechanism of the disease through genes related to cell proliferation or thyroid function. However, trisomies 7 and 10 have been reported in short-term cultures of nonneoplastic cells from patients with cancer [12, 13]; these findings may represent genomic instability in the tumor parenchyma or may characterize stromal fibroblasts [14].
This study was supported by Conselho Nacional de Desenvolvimento Cientifico e Tecnol6gico (CNPq), Funda§~o para o Desenvolvimento da UNESP (FUNDUNESP), Funda§8o de Amparo Pesqnisa do Estado de $8o Paulo (FAPESP) and Coordena~8o de Aperfei~oamento do Pessoal de Ensino Superior (CAPES). The authors thank Josu6 Rodrigues dos Santos and Rosana Silistino de Souza for technical assistance, Dr. Marileila Varella-Garcia for helpful discussion, and Dr. Carlos Daghlian for reviewing the text.
C y t o g e n e t i c s of M u l t i n o d u l a r T h y r o i d G o i t e r
77
REFERENCES 1. Rosai J (1989): Ackerman's Surgical Pathology, Vol. 1, 7th ed., C.V. Mosby, St. Louis, pp. 391-447. 2. Limon J, Dal Cin P, Sandberg AA (1986): Application of long term collagenase disaggregation for the cytogenetic analysis of h u m a n solid tumors. Cancer Genet Cytogenet 23:305317. 3. Wurster-Hill DH, Noll WW, Bircher LY, Pettengill OS, Grizzle WA (1986): Cytogenetics of medullary carcinoma of the thyroid. Cancer Genet Cytogenet 20:247-253. 4. Antonini P, Venuat AM, Linares G, Caillou B, Berger R, Parmentier C (1989): A translocation (7;10)(q35;q21) in a differentiated papillary carcinoma of the thyroid. Cancer Genet Cytogenet 41:139-144. 5. Bondeson L, Bengtsson A, Bondeson AG, Dahlenfors R, Grimelius L, Wedell B, Mark J (1989): Chromosome studies in thyroid neoplasia. Cancer 64:680-685. 6. Herath J, Jenkins R, Hay I, Grant C, Dewald G (1989): Cytogenetic studies on 12 thyroid carcinomas [Abstract]. In: International Workshop on Chromosomes in Solid Tumors, 3, Tucson, AZ, February 26 to March 3, 1989. Tucson, The Arizona Cancer Center and The National Cancer Institute, p. 64. 7. Herrmann ME, Bullerdiek J, Bartnitzke S, Lebert B, Lobeck H, Zuschneid W (1989): Cytogenetic studies on 15 thyroid adenomas [Abstract]. In: International Workshop on Chromosomes in Solid Tumors, 3, Tucson, AZ, February 26 to March 3, 1989. Tucson, The Arizona Cancer Center and The National Cancer Institute, p. 65. 8. Mitelman F (1985): Catalog of Chromosome Aberrations in Cancer, Vol. 5, 2nd ed. Alan R. Liss, New York, pp. 1-707. 9. Davies RL, Grosse VA, Kucherlapati R, Bothwell M (1980): Genetical analysis of epidermal growth factor action: Assignment of h u m a n epidermal growth factor receptor gene to chromosome 7. Proc Natl Acad Sci USA 77:4188-4192. 10. Smith M, Simpson NE (1989): Report of the committee on the genetic constitution of chromosomes 9 and 10. Cytogenet Cell Genet 51:202-225. 11. Miyajima N, Horiuchi R, Shibuya Y, Fukushige S, Matsubara K, Toyoshima K, Yamamoto T (1989): Two erbA homologs encoding proteins with different T3 binding capacities are transcribed from opposite DNA strands of the same genetic locus. Cell 57:31-39. 12. Heim S, Lindstrom E, Salford, LG (1990): Trisomy 7 and sex chromosome loss are not tumor-specific aberrations in malignant gliomas. Abstracts of 2nd European Workshop on Cytogenetics and Molecular Genetics of Human Solid Tumours: A18. 13. Szucs S, Emanuel A, Kovacs G (1990): Clonal aberrations of chromosomes X, Y, 7, and 10 in normal kidney tissue of patients with renal cell cancers. Abstracts of 2nd European Workshop on Cytogenetics and Molecular Genetics of Human Solid Tumours: P40. 14. Bardi G, Johansson B, Orndal C, Heim S, Mandahl N, Pandis N, Andren-Sandberg A, Mitelman F (1990): Abstracts of 2nd European Workshop on Cytogenetics and Molecular Genetics of Human Solid Tumours: P22.