Cytogenetic Study of a Pineocytoma Claudia Aparecida Rainho, Silvia Regina Rogatto, Lamartine Correa de Moraes, and Jos Barbieri-Neto
ABSTRACT: The cytogenetic findings based on G-banding in a pineocytoma detected in a 29-year-old
woman are reported. The chromosomal study showed numerical alterations involving c h r o m o s o m e s X, 5, 8, 11, 14, and 22, structural alterations of chromosomes 1, 3, 12, and 22, as well as various markers. Tumors of the pineal region are infrequent, and this is the first report of a p i n e o c y t o m a studied cytogenetically.
INTRODUCTION Primary tumors of the nervous system occur at a frequency estimated at 4.2-5.4/100,000 individuals and cause approximately 2.7% of all cancer deaths. Tumors of the pineal region are relatively rare. In the 0-20-year age range, they represent approximately 1.5%-3% of all nervous system tumors, and they practically do not occur in older individuals [1, 2]. The tumors originating in the parenchyma of the pineal gland are classified into two categories, i.e., pineoblastomas and pineocytomas. Pineoblastomas are primitive neuroectodermal tumors with poorly differentiated cells resembling medulloblastomas, whereas pineocytomas present a lobular pattern similar to that of the normal gland. Pineocytomas are benign tumors that spread slowly and have a better prognosis than all other tumors of the pineal region [3]. Nonrandom cytogenetic abnormalities have been described in a large number and in many types of human tumors. Among tumors of the nervous system, meningiomas have been studied most extensively [4-9]. Only three cases of pineoblastomas have been reported after cytogenetic evaluation, which revealed structural alterations involving chromosomes 1 and 11 [10, 11]. In the present report, we describe the cytogenetic analysis of cells from a pineocytoma detected in a 29-year-old woman.
diplopy starting 2 weeks before led her to seek our service. Neurologic examination showed a parinaud signal, bilateral papilledema, and a reduced level of consciousness with strong somnolence. A computerized tomography (CT) scan of the skull revealed a voluminous tumor in the pineal region expanding superiorly and anteriorly. The patient was submitted to microsurgery, and a frozen biopsy taken during the operation revealed the presence of a pineocytoma. The tumor was totally removed and histology confirmed the results of the frozen biopsy. The patient's postoperative course was good until the sixth day, when she died of a pulmonary embolus while deambulating in her room for physiotherapy. All attempts at resuscitation failed. CT scan control was not performed because of the unexpected evolution of the patient. Histologically the tumor consisted of cells of small volume, with scarce and ill-defined cytoplasm, hyperchromatic nuclei, and inconspicuous nucleoli. More voluminous cells with large and light cytoplasm containing a nucleus of finer chromatin and a small nucleolus appeared in association with the cells described previously. These cells formed clusters separated by incomplete septa supporting blood vessels, as observed in the normal pineal gland. The central region of the clusters showed cell necrosis frequently accompanied by central calcification. Mitotic figures were easily detected (Figs. 1 and 2).
CASE REPORT
A 29-year-old woman with a history of headache of many years' duration recently started to experience visual difficulties, being unable to lift her eyes up. Vomiting and
From the Departamento de Biologia Geral (C. A. R., S. R. R.), CCB, Departamento de Neurologia (L. C. I. M.), HU, Universidade Estadual de Londrina, PR, and Departamento de Patologia (J. B.-N.), FMRP, Universidade de S~o Paulo, SP, Brazil. Address reprint requests to: Silvia Regina Rogatto, PhD, Departamento de Biologia Geral, CCB, Universidade Estadual de Londrina, CEP 86051, Londrina, PR, Brazil. Received February 18, 1992; accepted June 5, 1992.
MATERIALS A N D METHODS
Fresh tumor tissue samples were obtained under sterile conditions and promptly processed. The fragments were first sectioned into small pieces and then enzymatically dissociated with a 0.4% solution of type IV collagenase (Sigma) and transferred to culture flasks containing Ham F-10 medium (Sigma) supplemented with 20% fetal calf serum, vitamins (DIFCO), and antibiotics. The cultures were kept at 37°C and fed twice a week. Culture time was determined individually for each flask depending on mitotic activity (20-28 days). For cytogenetic analysis, cells in the exponential growth phase were initially treated with 127
© 1992 Elsevier Science Publishing Co., Inc. 655 A v e n u e of the Americas, New York, NY 10010
Cancer Genet Cytogenet 64:127-132 (1992)
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Figure 1 Histologic structure of neoplastic tissue showing the connective septa between which the cells form clusters, with cells predominating here (HE x 360). 0.0016% colchicine for 6 hours and then with hypotonic 0.075 M KC1 for 30 minutes and finally fixed in methanol : acetic acid (3 : 1). Slides were subjected to standard Giemsa staining and to GTG b a n d i n g [12]. Karyotype description followed the nomenclature r e c o m m e n d e d by the ISCN 1985 [13]. RESULTS
Standard analysis of 100 cells revealed a m o d a l chromosome n u m b e r of 46 (63%), with numbers ranging from 37 to 99 chromosomes.
Thirty cells were evaluated after G-banding, and their karyotype formulae are presented in Table 1. Normal karyotypes were detected in nine cells. Table 2 s u m m a r i z e s the structural and numerical c h r o m o s o m e alterations observed and their respective frequencies. The most frequently detected chromosome alteration was m o n o s o m y of chromosome 22 (total: 26.7%; partial: 3.0%) (Figs. 3 and 4). Trisomy of chromosome 5 was observed in 10% of the cells. (Fig. 5). The structural c h r o m o s o m e alterations were n o n c l o n a l and i n c l u d e d inv(1)(p31q44) and deletions involving chromosomes 3, 12, and 22. Various c h r o m o s o m e markers were detected in eight cells (Fig. 4).
Figure 2 Neoplastic tissue mainly consisting of cells with ample and light cytoplasm usually arranged at the periphery of the lobule and of cells with regressive alterations in their center (HE x 480).
C y t o g e n e t i c S t u d y of a P i n e o c y t o m a
Table 1
129
K a r y o t y p e f o r m u l a e of 30 c e l l s after G - b a n d i n g
Table 2
37,X,-X,+5,-8,-9,-11,-12,-13,-16,-20,-21,-22 40,XX,- 3, + 5 , - 9 , - 12, - 14, - 1 7 , - 18, - 20 41,XX,- 6 , - 8 , - 9 , - 1 4 , - 22 42,XX,- 3 , - 8 , - 2 1 , - 22 4 3 , X , - X , - 1 1 , - 21 43,XX,- 1 7 , - 1 9 , - 22 44,XX,del(3)(q11),- 1 1 , - 17 45,XX,del(12)(q22?),- 21 45,XX, - 4 45,XX, - 14 45,XX, - 18 45,XX, - 22 45,XX, - 11, - 22, + mar3 45,XX, 2 1 , - 22,+ marl 46,XX ~ 46,X, - X, + 5, - 15, + marl 46,XX, - 16, + marl 46,XX,del(22)(q11.2) 46,XX, - 8, + mar2 46,XX, - 15, + mar4 46,XX, - 22, + marl 46,XX, + inv(1)(p31q44), - 6, - 8, - 16, + 22, + mar3
Chromosome alterations
Frequency (%)
Numeric -22 -8,-9,- 11,-21 -X,+5,-14 Structural inv(1)(p31q44) del(3)(q11) del(12)(q22?) del(22)(q11.2) Markers marl mar3 mar2, mar4
26,7 13,3 10,0 3.0 3.0 3.0 3.0 13.3 6.7 3.3
eries p r o v i d e d c o n v i n c i n g e v i d e n c e for t h e f u n d a m e n t a l role of c h r o m o s o m e a l t e r a t i o n s i n t h e m a l i g n a n t p r o c e s s [14]. M o r e r e c e n t l y , t h e d e f i n i t i o n of s o m e of t h e s e c h r o m o s o m e a b n o r m a l i t i e s at t h e m o l e c u l a r l e v e l led to t h e i d e n t i fication of c e r t a i n g e n e s t h a t p l a y a n i m p o r t a n t role in t u m o r t r a n s f o r m a t i o n a n d p r o g r e s s i o n [15]. R e p o r t s of c y t o g e n e t i c s t u d i e s o n t u m o r s of t h e p i n e a l r e g i o n are e x t r e m e l y rare. W e o n l y k n o w of t h r e e c a s e s of p i n e o b l a s t o m a r e p o r t e d t h u s far: t w o b y Griffin et al. [10], w h o d e s c r i b e d a l t e r a t i o n s i n v o l v i n g c h r o m o s o m e i as w e l l
o Only this karyotype was present as a second clone in nine cells; all the others were single karyotypes.
DISCUSSION C y t o g e n e t i c s t u d i e s p e r f o r m e d o v e r t h e last t w o d e c a d e s h a v e r e v e a l e d d i f f e r e n t t y p e s of c h r o m o s o m e a b n o r m a l i t i e s a s s o c i a t e d w i t h a large n u m b e r of n e o p l a s i a s . T h e s e d i s c o v -
Figure 3
S u m m a r y of c y t o g e n e t i c a b n o r m a l i t i e s a n d t h e i r frequencies
Cell with a 4 5 , X X , - 2 1 , - 2 2 , + marl karyotype
2
6
13
7
8
14
3
9
4
10
11
12
17
16
15
,,,,,0 19
mar 1
20
21
5
22
X
18
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A
3
D
C
22
12
mar I
mar 2
mar 3
mar 4
Figure 4 Partial karyotypes showing A. inv (1)(p31q64), B. del(3)(q11), C. del(12)(q22?), D. del(22)(q11.2) and E. chromosome markers (marl, mar2, mar3, and mar4).
as random chromosome losses, and a more recently described case involving del (11q) [11]. These reports refer to tumors of the pineal region detected in children up to 12 years of age and differ both in histologic pattern and in clinical progression from pineocytomas, which are more frequently detected in adults [16]. Some investigators have suggested differences in chromosome abnormalities between the general groups of tumors detected in children and in adults [10]. Comparison of our cytogenetic findings for the pineocytomas reported here with the cases of pineoblastoma described by the previous authors shows involvement of chromosome 1 in all cases: del(1)(p13p21), der(1)t(1;?), der(1)t(1;?)(q44;?) [10], and del(1)(p31p36) [11]. In our analysis, we identified inv(1)(p31q44) in a sporadic cell. The lp31 breakpoint was also involved in the
del(1) detected in only one cell by Sreekantaiah et al. [11]. This breakpoint coincides with the FRAIC fragile site [17] and with the location of oncogene JUN (AA Sandberg, personal communication). Chromosome 1 is involved in a large number of tumors [18] and may possibly be related to tumor progression, representing a secondary chromosome alteration. The del(11)(q13.1q13.5) also reported by Sreekantaiah et al. [11] may have a phenotypic effect comparable to total m o n o s o m y of chromosome 11 detected in our study. As suggested by these authors [11], chromosome 11 may be of etiologic importance in tumors of the pineal region, because, despite the low frequency of chromosome 11 m o n o s o m y observed in the present study, the complex chromosome picture suggests the presence of secondary chromosome alterations that progressively replaced the pri-
Cytogenetic Study of a Pineocytoma
131
3
I
4
YP
lllbm 6
7
8
9
10
12
11
X
0 13
14
15
16
17
18
o.
19
mar
20
21
22
1
Figure 5
The arrows indicate trisomy of chromosome 5, monosomy of chromosomes X and 15 and marl.
mary alteration [19]. Total or partial monosomy of chromosome 11 has been described in several types of cancer, and its significance in terms of the loss of heterozygosity of genes located on it has been investigated [20]. The cytogenetic alteration most frequently detected in our analysis was m o n o s o m y of chromosome 22, which is a chromosome abnormality considered to be specific and consistent in m e n i n g i o m a s [4-9]. Total or partial monosomy of chromosome 22 is considered to be the primary n o n r a n d o m event, progressively leading to the genesis of lines with other structural or numerical rearrangements [21]. Some investigators have suggested a n o n r a n d o m pattern of alterations secondary to m o n o s o m y of chromosome 22 in meningiomas, m a i n l y involving chromosomes 8, 9, 14, and 15 and gonosomes for numerical deviations [22] and chromosome i and 22 for structural rearrangements [7, 8]. It is important to emphasize the great similarity between our data and this cytogenetic pattern described for meningiomas. In addition to 22 monosomy, we reported monosomy of chromosomes X, 8, 9, and 14, as well as structural alterations of chromosomes 1 and 22. The large n u m b e r of chromosome losses and structural rearrangements described in the present case, i n c l u d i n g trisomy of chromosome 5, suggests a more evolved picture w i t h i n a multiple-stage neoplastic process [23]. Probably chromosome 22 plays a general role in the etiology of various tumors of the nervous system [24, 25], i n c l u d i n g those of the pineal region. There is an obvious need for further study of tumors
of the pineal region. Molecular analysis associated with cytogenetic data may help elucidate specific chromosome abnormalities and identify relevant genes in neoplasias of the pineal gland, especially with respect to the role of chromosomes 1, 11, and 22.
The authors wish to thank the director and staff of Santa Casa de Miseric6rdia de Londrina, and Dr. Ronaldo Vasconcelos for permitting access to slides for histopathologic reevaluation. This research was financed by CNPq, CAPES, and CPG-Universidade Estadual de Londrina, PR, Brazil.
REFERENCES 1. Youmans JR (1980): Tumors of Pineal Region. In: Neurological Surgery--A Comprehensive Reference Guide to the Diagnosis and Management of Neurosurgical Problems, Vol. 5, 2nd Ed., Saunders, Philadelphia, pp. 2863-2871. 2. Walker MD (1982): Brain and peripheral nervous system tumors. In: Cancer medicine, 2nd Ed., JF Holland, E Frei III, eds. Lea & Febiger, Philadelphia, pp. 1603-1633. 3. Rorke LB, Gilles FH, Davis RL, Becker LE (1985): Revision of the World Health Organization classification of brain tumors for childhood brain tumors. Cancer 56:1869-1886. 4. Mark J (1973): Karyotype patterns in human meningiomas: A comparison between studies with G and Q banding techniques. Hereditas 75:213-220. 5. Zankl H, Zang KD (1980): Correlations between clinical and cytogenetical data in 180 human meningiomas. Cancer Genet Cytogenet 1:351-356.
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6. Maltby EL, Ironside JW, Battersbyrde (1988): Cytogenetic studies in 50 meningiomas. Cancer Genet Cytogenet 31:199-210. 7. Rogatto SR, Casartelli C (1988): Cytogenetic analysis of h u m a n meningiomas. Rev Brasil Genet 11:729-744. 8. Casartelli C, Rogatto SR, Barbieri Neto J. (1989): Karyotypic evolution of h u m a n meningioma: Progression through malignancy. Cancer Genet Cytogenet 40:33-45. 9. Casalone R, Simi P, Granata P. Minelli E, Giudici A, Butti G, Solero CL (1990): Correlation between cytogenetic and histopathological findings in 65 h u m a n meningiomas. Cancer Genet Cytogenet 45:237-243. 10. Griffin CA, Hawkins AL, Packer RJ, Rorke LB, Emanuel BS (1988): Chromosome abnormalities in pediatric brain tumors. Cancer Res 48:175-180. 11. Sreekantaiah C, Jokin H, Brecher ML, Sandberg AA (19891: Interstitial deletion of chromosome 11q in a pineoblastoma. Cancer Genet Cytogenet 39:125-131. 12. Scheres VMJC (1972): Identification of two Robertsonian translocation with a Giemsa banding technique. Hum Genet 15:253-256. 13. ISCN (1985): An International System for Human Cytogenetic Nomenclature, Harnden DG, Klinger HP (eds.): published in collaboration with Cytogenet Ceil Genet (Karger, Basel, 1985); also in Birth Defects: Original Article Series, Vol. 21, No. 1 (March of Dimes Birth Defects Foundation, New York, 1985). 14. Mitelman F (1986): Clustering of breakpoints to specific chromosomal regions in h u m a n neoplasia. A survey of 5,345 cases. Hereditas 104:113-119. 15. Cavenee WK, Scrable HJ, James CD (1991). Molecular genetics
C . A . R a i n h o et al.
16.
17.
18. 19. 20.
21.
22.
23. 24.
25.
of h u m a n cancer predisposition and progression. Mutat Res 247:199-202 De Vita VT, Hellman S, Rosenberg SA, eds (1982): Cancer: Principles and Practice on Oncology. Lippincott, Philadelphia. Hecht F, Ramesh KH, Lockwood DH (1990): A guide to fragile sites on h u m a n chromosomes. Cancer Genet Cytogenet 44:37-45. Mitelman F (1988): Catalog of Chromosome Aberrations in Cancer, 3rd Ed. Alan R. Liss, New York. Heim S, Mitelman F (1989): Primary chromosome abnormalities in h u m a n neoplasia. Adv Cancer Res 52:1-43. Atkin NB, Baker MC (1988). Deficiency of all or part of chromosome 11 in several types of cancer: Significance of a reduction in the number of normal chromosomes 11. Cytogenet Cell Genet 47:106-107. Rey JA, Bello JM, Campos JM, Kusar E, Moreno S (1988): Chromosomal involvement secondary to - 2 2 in h u m a n meningiomas. Cancer Genet Cytogenet 33:275-290. Mark J (1977): Chromosomal abnormalities and their specificity in h u m a n neoplasms: An assessment of recent observations by banding techniques. Adv Cancer Res 24:165-222. Nowell PC (1986): Mechanisms of tumor progression. Cancer Res 46:2203-3307. Yamada K, Kondo T, Yoshioka M, Oami H (1980): Cytogenetic studies in twenty h u m a n brain tumors: Association of no. 22 chromosome abnormalities with tumors of the brain. Cancer Genet Cytogenet 2:293-307. Bigner SH, Mark J, Bigner DD (1990): Cytogenetics of h u m a n brain tumors. Cancer Genet Cytogenet 47:141-154.
ABSTRACT: The cytogenetic findings based on G-banding in a pineocytoma detected in a 29-year-old
woman are reported. The chromosomal study showed numerical alterations involving c h r o m o s o m e s X, 5, 8, 11, 14, and 22, structural alterations of chromosomes 1, 3, 12, and 22, as well as various markers. Tumors of the pineal region are infrequent, and this is the first report of a p i n e o c y t o m a studied cytogenetically.
INTRODUCTION Primary tumors of the nervous system occur at a frequency estimated at 4.2-5.4/100,000 individuals and cause approximately 2.7% of all cancer deaths. Tumors of the pineal region are relatively rare. In the 0-20-year age range, they represent approximately 1.5%-3% of all nervous system tumors, and they practically do not occur in older individuals [1, 2]. The tumors originating in the parenchyma of the pineal gland are classified into two categories, i.e., pineoblastomas and pineocytomas. Pineoblastomas are primitive neuroectodermal tumors with poorly differentiated cells resembling medulloblastomas, whereas pineocytomas present a lobular pattern similar to that of the normal gland. Pineocytomas are benign tumors that spread slowly and have a better prognosis than all other tumors of the pineal region [3]. Nonrandom cytogenetic abnormalities have been described in a large number and in many types of human tumors. Among tumors of the nervous system, meningiomas have been studied most extensively [4-9]. Only three cases of pineoblastomas have been reported after cytogenetic evaluation, which revealed structural alterations involving chromosomes 1 and 11 [10, 11]. In the present report, we describe the cytogenetic analysis of cells from a pineocytoma detected in a 29-year-old woman.
diplopy starting 2 weeks before led her to seek our service. Neurologic examination showed a parinaud signal, bilateral papilledema, and a reduced level of consciousness with strong somnolence. A computerized tomography (CT) scan of the skull revealed a voluminous tumor in the pineal region expanding superiorly and anteriorly. The patient was submitted to microsurgery, and a frozen biopsy taken during the operation revealed the presence of a pineocytoma. The tumor was totally removed and histology confirmed the results of the frozen biopsy. The patient's postoperative course was good until the sixth day, when she died of a pulmonary embolus while deambulating in her room for physiotherapy. All attempts at resuscitation failed. CT scan control was not performed because of the unexpected evolution of the patient. Histologically the tumor consisted of cells of small volume, with scarce and ill-defined cytoplasm, hyperchromatic nuclei, and inconspicuous nucleoli. More voluminous cells with large and light cytoplasm containing a nucleus of finer chromatin and a small nucleolus appeared in association with the cells described previously. These cells formed clusters separated by incomplete septa supporting blood vessels, as observed in the normal pineal gland. The central region of the clusters showed cell necrosis frequently accompanied by central calcification. Mitotic figures were easily detected (Figs. 1 and 2).
CASE REPORT
A 29-year-old woman with a history of headache of many years' duration recently started to experience visual difficulties, being unable to lift her eyes up. Vomiting and
From the Departamento de Biologia Geral (C. A. R., S. R. R.), CCB, Departamento de Neurologia (L. C. I. M.), HU, Universidade Estadual de Londrina, PR, and Departamento de Patologia (J. B.-N.), FMRP, Universidade de S~o Paulo, SP, Brazil. Address reprint requests to: Silvia Regina Rogatto, PhD, Departamento de Biologia Geral, CCB, Universidade Estadual de Londrina, CEP 86051, Londrina, PR, Brazil. Received February 18, 1992; accepted June 5, 1992.
MATERIALS A N D METHODS
Fresh tumor tissue samples were obtained under sterile conditions and promptly processed. The fragments were first sectioned into small pieces and then enzymatically dissociated with a 0.4% solution of type IV collagenase (Sigma) and transferred to culture flasks containing Ham F-10 medium (Sigma) supplemented with 20% fetal calf serum, vitamins (DIFCO), and antibiotics. The cultures were kept at 37°C and fed twice a week. Culture time was determined individually for each flask depending on mitotic activity (20-28 days). For cytogenetic analysis, cells in the exponential growth phase were initially treated with 127
© 1992 Elsevier Science Publishing Co., Inc. 655 A v e n u e of the Americas, New York, NY 10010
Cancer Genet Cytogenet 64:127-132 (1992)
0165-4608/92/$05.00
128
C . A . Rainho et al.
Figure 1 Histologic structure of neoplastic tissue showing the connective septa between which the cells form clusters, with cells predominating here (HE x 360). 0.0016% colchicine for 6 hours and then with hypotonic 0.075 M KC1 for 30 minutes and finally fixed in methanol : acetic acid (3 : 1). Slides were subjected to standard Giemsa staining and to GTG b a n d i n g [12]. Karyotype description followed the nomenclature r e c o m m e n d e d by the ISCN 1985 [13]. RESULTS
Standard analysis of 100 cells revealed a m o d a l chromosome n u m b e r of 46 (63%), with numbers ranging from 37 to 99 chromosomes.
Thirty cells were evaluated after G-banding, and their karyotype formulae are presented in Table 1. Normal karyotypes were detected in nine cells. Table 2 s u m m a r i z e s the structural and numerical c h r o m o s o m e alterations observed and their respective frequencies. The most frequently detected chromosome alteration was m o n o s o m y of chromosome 22 (total: 26.7%; partial: 3.0%) (Figs. 3 and 4). Trisomy of chromosome 5 was observed in 10% of the cells. (Fig. 5). The structural c h r o m o s o m e alterations were n o n c l o n a l and i n c l u d e d inv(1)(p31q44) and deletions involving chromosomes 3, 12, and 22. Various c h r o m o s o m e markers were detected in eight cells (Fig. 4).
Figure 2 Neoplastic tissue mainly consisting of cells with ample and light cytoplasm usually arranged at the periphery of the lobule and of cells with regressive alterations in their center (HE x 480).
C y t o g e n e t i c S t u d y of a P i n e o c y t o m a
Table 1
129
K a r y o t y p e f o r m u l a e of 30 c e l l s after G - b a n d i n g
Table 2
37,X,-X,+5,-8,-9,-11,-12,-13,-16,-20,-21,-22 40,XX,- 3, + 5 , - 9 , - 12, - 14, - 1 7 , - 18, - 20 41,XX,- 6 , - 8 , - 9 , - 1 4 , - 22 42,XX,- 3 , - 8 , - 2 1 , - 22 4 3 , X , - X , - 1 1 , - 21 43,XX,- 1 7 , - 1 9 , - 22 44,XX,del(3)(q11),- 1 1 , - 17 45,XX,del(12)(q22?),- 21 45,XX, - 4 45,XX, - 14 45,XX, - 18 45,XX, - 22 45,XX, - 11, - 22, + mar3 45,XX, 2 1 , - 22,+ marl 46,XX ~ 46,X, - X, + 5, - 15, + marl 46,XX, - 16, + marl 46,XX,del(22)(q11.2) 46,XX, - 8, + mar2 46,XX, - 15, + mar4 46,XX, - 22, + marl 46,XX, + inv(1)(p31q44), - 6, - 8, - 16, + 22, + mar3
Chromosome alterations
Frequency (%)
Numeric -22 -8,-9,- 11,-21 -X,+5,-14 Structural inv(1)(p31q44) del(3)(q11) del(12)(q22?) del(22)(q11.2) Markers marl mar3 mar2, mar4
26,7 13,3 10,0 3.0 3.0 3.0 3.0 13.3 6.7 3.3
eries p r o v i d e d c o n v i n c i n g e v i d e n c e for t h e f u n d a m e n t a l role of c h r o m o s o m e a l t e r a t i o n s i n t h e m a l i g n a n t p r o c e s s [14]. M o r e r e c e n t l y , t h e d e f i n i t i o n of s o m e of t h e s e c h r o m o s o m e a b n o r m a l i t i e s at t h e m o l e c u l a r l e v e l led to t h e i d e n t i fication of c e r t a i n g e n e s t h a t p l a y a n i m p o r t a n t role in t u m o r t r a n s f o r m a t i o n a n d p r o g r e s s i o n [15]. R e p o r t s of c y t o g e n e t i c s t u d i e s o n t u m o r s of t h e p i n e a l r e g i o n are e x t r e m e l y rare. W e o n l y k n o w of t h r e e c a s e s of p i n e o b l a s t o m a r e p o r t e d t h u s far: t w o b y Griffin et al. [10], w h o d e s c r i b e d a l t e r a t i o n s i n v o l v i n g c h r o m o s o m e i as w e l l
o Only this karyotype was present as a second clone in nine cells; all the others were single karyotypes.
DISCUSSION C y t o g e n e t i c s t u d i e s p e r f o r m e d o v e r t h e last t w o d e c a d e s h a v e r e v e a l e d d i f f e r e n t t y p e s of c h r o m o s o m e a b n o r m a l i t i e s a s s o c i a t e d w i t h a large n u m b e r of n e o p l a s i a s . T h e s e d i s c o v -
Figure 3
S u m m a r y of c y t o g e n e t i c a b n o r m a l i t i e s a n d t h e i r frequencies
Cell with a 4 5 , X X , - 2 1 , - 2 2 , + marl karyotype
2
6
13
7
8
14
3
9
4
10
11
12
17
16
15
,,,,,0 19
mar 1
20
21
5
22
X
18
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A
3
D
C
22
12
mar I
mar 2
mar 3
mar 4
Figure 4 Partial karyotypes showing A. inv (1)(p31q64), B. del(3)(q11), C. del(12)(q22?), D. del(22)(q11.2) and E. chromosome markers (marl, mar2, mar3, and mar4).
as random chromosome losses, and a more recently described case involving del (11q) [11]. These reports refer to tumors of the pineal region detected in children up to 12 years of age and differ both in histologic pattern and in clinical progression from pineocytomas, which are more frequently detected in adults [16]. Some investigators have suggested differences in chromosome abnormalities between the general groups of tumors detected in children and in adults [10]. Comparison of our cytogenetic findings for the pineocytomas reported here with the cases of pineoblastoma described by the previous authors shows involvement of chromosome 1 in all cases: del(1)(p13p21), der(1)t(1;?), der(1)t(1;?)(q44;?) [10], and del(1)(p31p36) [11]. In our analysis, we identified inv(1)(p31q44) in a sporadic cell. The lp31 breakpoint was also involved in the
del(1) detected in only one cell by Sreekantaiah et al. [11]. This breakpoint coincides with the FRAIC fragile site [17] and with the location of oncogene JUN (AA Sandberg, personal communication). Chromosome 1 is involved in a large number of tumors [18] and may possibly be related to tumor progression, representing a secondary chromosome alteration. The del(11)(q13.1q13.5) also reported by Sreekantaiah et al. [11] may have a phenotypic effect comparable to total m o n o s o m y of chromosome 11 detected in our study. As suggested by these authors [11], chromosome 11 may be of etiologic importance in tumors of the pineal region, because, despite the low frequency of chromosome 11 m o n o s o m y observed in the present study, the complex chromosome picture suggests the presence of secondary chromosome alterations that progressively replaced the pri-
Cytogenetic Study of a Pineocytoma
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Figure 5
The arrows indicate trisomy of chromosome 5, monosomy of chromosomes X and 15 and marl.
mary alteration [19]. Total or partial monosomy of chromosome 11 has been described in several types of cancer, and its significance in terms of the loss of heterozygosity of genes located on it has been investigated [20]. The cytogenetic alteration most frequently detected in our analysis was m o n o s o m y of chromosome 22, which is a chromosome abnormality considered to be specific and consistent in m e n i n g i o m a s [4-9]. Total or partial monosomy of chromosome 22 is considered to be the primary n o n r a n d o m event, progressively leading to the genesis of lines with other structural or numerical rearrangements [21]. Some investigators have suggested a n o n r a n d o m pattern of alterations secondary to m o n o s o m y of chromosome 22 in meningiomas, m a i n l y involving chromosomes 8, 9, 14, and 15 and gonosomes for numerical deviations [22] and chromosome i and 22 for structural rearrangements [7, 8]. It is important to emphasize the great similarity between our data and this cytogenetic pattern described for meningiomas. In addition to 22 monosomy, we reported monosomy of chromosomes X, 8, 9, and 14, as well as structural alterations of chromosomes 1 and 22. The large n u m b e r of chromosome losses and structural rearrangements described in the present case, i n c l u d i n g trisomy of chromosome 5, suggests a more evolved picture w i t h i n a multiple-stage neoplastic process [23]. Probably chromosome 22 plays a general role in the etiology of various tumors of the nervous system [24, 25], i n c l u d i n g those of the pineal region. There is an obvious need for further study of tumors
of the pineal region. Molecular analysis associated with cytogenetic data may help elucidate specific chromosome abnormalities and identify relevant genes in neoplasias of the pineal gland, especially with respect to the role of chromosomes 1, 11, and 22.
The authors wish to thank the director and staff of Santa Casa de Miseric6rdia de Londrina, and Dr. Ronaldo Vasconcelos for permitting access to slides for histopathologic reevaluation. This research was financed by CNPq, CAPES, and CPG-Universidade Estadual de Londrina, PR, Brazil.
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