Constitutional Balanced Translocations in Patients with Solid Tumors Florence Richard, Martine Muleris, J r6me Couturier, Mich le Gerbault-Seureau, Martine Lombard, and Bernard Dutrillaux
ABSTRACT: In a sample of 329 patients with a solid t u m o r (colon and breast adenocarcinoma, cervical carcinoma, a n d meningioma), four balanced constitutional translocations were observed. Two were t(13q14q), and two were reciprocal translocations. Comparison with surveys of newborns showed a significant excess of translocations in our sample.
INTRODUCTION
MATERIALS AND METHODS
A p p r o x i m a t e l y 1 of 150 n e w b o r n s has a constitutional c h r o m o s o m e a n o m a l y w h i c h consists of u n b a l a n c e d autosomal aberrations, u n b a l a n c e d sex chromosome aberrations, and balanced structural rearrangements, in roughly equal proportions [1]. Some constitutional c h r o m o s o m e abnormalities determine an increased risk of malignancy: del(11p) is associated with an elevated risk for W i l m s ' tumor, del(13q) is associated with an elevated risk for retinoblastoma, trisomy 21 is associated with a m a r k e d l y elevated risk of l e u k e m i a [2], and del(5q) is related to a d e n o m a t o u s p o l y p o s i s coli (APC) s y n d r o m e [3, 4]. Several case reports have described simultaneous occurrence of other constitutional c h r o m o s o m a l aberrations and cancer [2], but the true extent of such associations or whether they represent r a n d o m events is not yet known. The only studies of the i n c i d e n c e of congenital c h r o m o s o m e anomalies and n e o p l a s i a in large series were made by A l i m e n a et al. [5] and Benitez et al. [6], who studied patients with hematologic disorders. Although the frequency of congenital c h r o m o s o m e alterations other than Down s y n d r o m e in their s a m p l e was twice that of normal population, Benitez et al. c o n c l u d e d that this difference was not statistically significant [6]. Very little is k n o w n about the frequency of constitutional chromosomal anomaly in i n d i v i d u a l s with solid tumors. Having observed constitutional structural rearrangements in four such patients, we believed it of interest to describe our series which, although limited, suggests an excess of structural rearrangements in such patients.
Our sample comprises 106 patients w i t h colorectal adenocarcinoma i n c l u d i n g 20 APC patients, 107 patients w i t h breast adenocarcinoma, 91 patients with c a r c i n o m a of the uterine cervix, and 25 patients w i t h meningioma. The constitutional karyotype was established in various circumstances: blood culture and analysis of 100 m e t a p h a s e s for a prospective s t u d y of c h r o m o s o m a l instability, b l o o d culture and analysis of a limited n u m b e r of metaphases, t u m o r culture p r o v i d i n g fibroblast growth, t u m o r culture providing cells with normal karyotype associated or not w i t h cells having abnormal karyotype, and t u m o r culture p r o v i d i n g cells with a single c h r o m o s o m a l anomaly, w i t h control on blood l y m p h o c y t e s to establish the existence of a congenital anomaly. All karyotypes were s t u d i e d after R-banding. In addition, some studies were p e r f o r m e d with 5 - b r o m o d e o x y u r i d i n e (BrdU) incorporation. All metaphases were k a r y o t y p e d on photographs.
From CNRS URA 620, Institut Curie, Section de Biologie, Paris, France. Address reprint requests to: Dr. F. Richard, CNRS URA 620, Institut Curie, Section de Biologie, Pavillon Trouillet-Rossignol, 26 rue d'Ulm, 75231 Paris C~dex 05, France. Received December 30, 1991; accepted January 23, 1992.
5O Cancer Genet Cytogenet 61:50-52 (1992) 0165-4608/92/$05.00
RESULTS AND DISCUSSION Structural Rearrangements One balanced translocation was detected in each s a m p l e corresponding to one t u m o r type (Table 1). Two were Robertsonian translocations 13q14q. T h e y were detected in prospective blood l y m p h o c y t e studies from a patient with non-APC colorectal a d e n o c a r c i n o m a and a patient with a breast a d e n o c a r c i n o m a (samples A and B, Table 1), i.e., among 104 patients unselected for a s u s p i c i o n of constitutional a n o m a l y from t u m o r material analysis. This provides an u p p e r estimate for i n c i d e n c e of constitutional chromosome alterations in our sample: two of 104 = 1.92%, but we obtained information on the constitutional karyotype of some patients retrospectively by analysis of t u m o r material that exhibited either normal karyotypes or karyotypes with a single structural a n o m a l y whose congenital origin could be confirmed by blood l y m p h o c y t e analyses. This increases the total sample to 329 (APC patients included), w h i c h yields the lower estimate of incidence: two of 329 : 0.6% © 1992 Elsevier Science Publishing Co. Inc. 655 Avenue of the Americas~New York. NY 10010
Constitutional Balanced Translocations in Solid Tumors
Table 1
51
Constitutional c h r o m o s o m e anomalies in adult patients with solid tumors
Population
Tissue sample (n)
Non-APC colorectal adenocarcinoma (n = 86)
A(13) B(44)
APC colorectal adenocarcinoma (n : 20) Breast adenocarcinoma (n ~ 107)
C(11] D(18) A(11) B (9) A(16) 9(31)
Cervical carcinoma (n = 91) Meningioma (n = 25)
Constitutional chromosomal aberration fra(10)(q24.2) t(13q14q) fra(2)(q22)
del(5) in two relatives fra(2)(q14.1) -t(13q14q) fra(10)(q25)
C(35) B(25) D(91)
t(3;15)(q13.3;q21)
D(25)
t(3;6)(q13.3;q22)
Abbreviation: APC, adenomatous polyposis coli; n, sample size.
Tissue sample A, 100 lymphocytemetaphases analyzed; tissue sample B, 2-65 lymphocytes metaphases analyzed; tissue sample C, only normal karyotypes observed in tumor biopsy; tissue sample D, normal karyotypes observed beside abnormal karyotypes in tumor biopsy. In addition, for samples C and D, the possible constitutional origin of simple balanced rearrangements was verified by study of blood lymphocytes.
of our cases, w h i c h represents a sixfold increase frequency of Robertsonian translocations as compared with that observed in the n e w b o r n p o p u l a t i o n (p ~ 0.01) (Table 2). The two other balanced rearrangements were reciprocal translocations. Both were detected after analysis of tumor material and retrospective study of lymphocytes. They represent two of 329 -- .6% of studied cases, w h i c h is also sixfold higher than that in the normal population (p 0.01). A m o n g colorectal cancers, APC patients were analyzed separately, because a deletion of 5q arm, in w h i c h the gene is m a p p e d [7, 8] may be a direct etiologic factor [3, 4]. In a sample of 20 patients from 17 i n d e p e n d e n t families, a del(5)(q14q23.1) was observed effectively in two relatives. W h e n APC patients w h o had a special ascertainm e n t are excluded, the total occurrence of constitutional structural rearrangements can be estimated as 1.22%. We cannot draw definite conclusions about this freq u e n c y because of the limited n u m b e r of patients. To obtain
Table 2
a better estimate of i n c i d e n c e of constitutional alterations in individuals with solid tumor, we r e v i e w e d data from the literature. To our knowledge, only two studies refer to significant series of constitutional karyotypes in adult patients affected by solid tumors. In a series of 50 patients with breast cancer, Ochi et al. [9] observed one reciprocal translocation. In another series of 76 cases of breast and 30 cases of lung cancer, Pathak et al. [10] did not report any constitutional c h r o m o s o m a l rearrangement. F r o m these data and ours, we calculated the f r eq u en cy of constitutional c h r o m o s o m e alteration in the p o p u l a t i o n w i t h solid t u m o r and compared it with the i n c i d e n c e at birth (Table 2). Although the sample of patients is still small, it confirms that a significant excess of constitutional balanced rearrangements exists in patients affected by a solid tumor. This excess may be sixfold (Table 2). In corollary, it ma y indicate a higher risk of cancer in i n d i v i d u a l s w i t h such rearrangements, although prospective studies of larger se-
Constitutional balanced c h r o m o s o m e rearrangements observed in the p o p u l a t i o n w i t h solid tumor and in newborns Population with solid tumor
Chromosomal anomaly
Our series (n = 329)
Total c (n = 455)
Robertsonian translocations Reciprocal translocations All translocations Inversions
2 (0.6)a 2 (0.6)° 4 (1.22) ~ 0
2 (0.44) a 3 (0.66) b 5 (1.1) b 0
Newborn population d (n ~ 67,014) 62 60 122 12
(0.09) (0.09) (0.18) (0.02)
n, sample size; percentages are indicated in brackets. Each sample was compared to newborns, and the probability of observingsuch distributions was calculated under a Poisson distribution: ° p ~ 0.01, bp ~ 0.001. ~:Includes our sample and the two published series of Ochi et al. [9] and Pathak et al. [10]. d UNSCEAR[1].
52
ries of patients are necessary before any definite conclusion can be reached. Again because of the small sample size, it would be premature to conclude that specific chromosome breakpoints exist, although chromosome 3 was involved twice in reciprocal translocation after breakage in band 3q13.3.
Constitutional Fragile Sites Our study was obviously not performed u n d e r optimal conditions to detect constitutional fragile sites because a limited n u m b e r of metaphases was studied in most cases. Furthermore, fragile sites are usually expressed in lymphocytes and not in tumor cells [10], and we had information on blood lymphocytes for 130 patients only. In this sample, we detected two cases of fra(10)(q24.2), one case of fra(2)(q14.1), and one case of fra(2)(q22). Another case of fra(10)(q25) was described by Ochi et al. [9] among 50 breast cancer patients; no information was provided by Pathak et al. [10]. This yields a frequency of 2.8% for the detected rare fragile sites, which is in the range of that of the normal population [11, 12], but our observed frequency probably is greatly underestimated and an increased frequency of fragile site carriers among patients with solid tumors cannot be excluded.
REFERENCES 1. UNSCEAR Report (1982): Ionizing radiation: Sources and biological effects. United Nations Scientific Committee on the Effects of Atomic Radiation, United Nations, New York. 2. Sandberg AA (1990): The Chromosomes in Human Cancer and Leukemia, 2nd Ed. Elsevier Science Publishing Co., New York.
F. Richard et al.
3. Herrera L, Kakati S, Gibas L, Pietrzak E, Sandberg AA (1986): Gardner's syndrome in a man with an interstitial deletion of 5q. Am J Med Genet 25:473-476. 4. Hockey KA, Mulcahy MT, Montgomery P, Levitt S (1989): Deletion of chromosome 5q and familial adenomatous polyposis. J Med Genet 26:61-62. 5. Alimena G, Billstr6m R, Casalone R, Gallo E, Mitelman F, Pasquali F (1985): Cytogenetic pattern in leukemic cells of patients with constitutional chromosome anomalies. Cancer Genet Cytogenet 16:207-218. 6. Benitez J, Valcarcel E, Ramos C, Ayuso C, Sanchez Cascos A (1987): Frequency of constitutional chromosome alterations in patients with hematologic neoplasias. Cancer Genet Cytogenet 24:345-354. 7. Bodmer WF, Bailey CJ, Bodmer J, Bussey HJR, Ellis A, Gorman P, Lucibello FC, Murday VA, Rider SH, Scambler P, Sheer D, Solomon E, Spurr NK (1987): Localization of the gene for familial adenomatous polyposis on chromosome 5. Nature 328:614-616. 8. Leppert M, Dobbs M, Scambler P, O'Connell P, Nakamura Y, Stauffer D, Wooward S, Burt R, Hughes J, Gardner E, Lathrop M, Wasmuth J, Lalovel JM, White R (1987): The gene for familial polyposis coli maps to the long arm of chromosome 5. Science 238:1411-1413. 9. Ochi H, Watanabe S, Furuya T, Tsugane S (1988): Chromosome fragility of lymphocytes from breast cancer patients in relation to epidemiologic data. Jpn J Cancer Res 79:1024-1030. 10. Pathak S, Hopwood VL, Hortobagyi GN, Jackson GL, Hughes JI, Melillo D (1991): Chromosome anomalies in human breast cancer: Evidence for specific involvement of lq region in lymphocyte cultures. Anticancer Res 11:1055-1060. 11. Muleris M, Dutrillaux AM, Lombard M, Dutrillaux B (1987): Noninvolvement of a constitutional heritable fragile site at 10q24.2 in rearranged chromosomes from rectal carcinoma cells. Cancer Genet Cytogenet 25:7-13. 12. Takahashi E, Hori T, Murata M (1988): Population cytogenetics of rare fragile sites in Japan. Hum Genet 78:121-126.
ABSTRACT: In a sample of 329 patients with a solid t u m o r (colon and breast adenocarcinoma, cervical carcinoma, a n d meningioma), four balanced constitutional translocations were observed. Two were t(13q14q), and two were reciprocal translocations. Comparison with surveys of newborns showed a significant excess of translocations in our sample.
INTRODUCTION
MATERIALS AND METHODS
A p p r o x i m a t e l y 1 of 150 n e w b o r n s has a constitutional c h r o m o s o m e a n o m a l y w h i c h consists of u n b a l a n c e d autosomal aberrations, u n b a l a n c e d sex chromosome aberrations, and balanced structural rearrangements, in roughly equal proportions [1]. Some constitutional c h r o m o s o m e abnormalities determine an increased risk of malignancy: del(11p) is associated with an elevated risk for W i l m s ' tumor, del(13q) is associated with an elevated risk for retinoblastoma, trisomy 21 is associated with a m a r k e d l y elevated risk of l e u k e m i a [2], and del(5q) is related to a d e n o m a t o u s p o l y p o s i s coli (APC) s y n d r o m e [3, 4]. Several case reports have described simultaneous occurrence of other constitutional c h r o m o s o m a l aberrations and cancer [2], but the true extent of such associations or whether they represent r a n d o m events is not yet known. The only studies of the i n c i d e n c e of congenital c h r o m o s o m e anomalies and n e o p l a s i a in large series were made by A l i m e n a et al. [5] and Benitez et al. [6], who studied patients with hematologic disorders. Although the frequency of congenital c h r o m o s o m e alterations other than Down s y n d r o m e in their s a m p l e was twice that of normal population, Benitez et al. c o n c l u d e d that this difference was not statistically significant [6]. Very little is k n o w n about the frequency of constitutional chromosomal anomaly in i n d i v i d u a l s with solid tumors. Having observed constitutional structural rearrangements in four such patients, we believed it of interest to describe our series which, although limited, suggests an excess of structural rearrangements in such patients.
Our sample comprises 106 patients w i t h colorectal adenocarcinoma i n c l u d i n g 20 APC patients, 107 patients w i t h breast adenocarcinoma, 91 patients with c a r c i n o m a of the uterine cervix, and 25 patients w i t h meningioma. The constitutional karyotype was established in various circumstances: blood culture and analysis of 100 m e t a p h a s e s for a prospective s t u d y of c h r o m o s o m a l instability, b l o o d culture and analysis of a limited n u m b e r of metaphases, t u m o r culture p r o v i d i n g fibroblast growth, t u m o r culture providing cells with normal karyotype associated or not w i t h cells having abnormal karyotype, and t u m o r culture p r o v i d i n g cells with a single c h r o m o s o m a l anomaly, w i t h control on blood l y m p h o c y t e s to establish the existence of a congenital anomaly. All karyotypes were s t u d i e d after R-banding. In addition, some studies were p e r f o r m e d with 5 - b r o m o d e o x y u r i d i n e (BrdU) incorporation. All metaphases were k a r y o t y p e d on photographs.
From CNRS URA 620, Institut Curie, Section de Biologie, Paris, France. Address reprint requests to: Dr. F. Richard, CNRS URA 620, Institut Curie, Section de Biologie, Pavillon Trouillet-Rossignol, 26 rue d'Ulm, 75231 Paris C~dex 05, France. Received December 30, 1991; accepted January 23, 1992.
5O Cancer Genet Cytogenet 61:50-52 (1992) 0165-4608/92/$05.00
RESULTS AND DISCUSSION Structural Rearrangements One balanced translocation was detected in each s a m p l e corresponding to one t u m o r type (Table 1). Two were Robertsonian translocations 13q14q. T h e y were detected in prospective blood l y m p h o c y t e studies from a patient with non-APC colorectal a d e n o c a r c i n o m a and a patient with a breast a d e n o c a r c i n o m a (samples A and B, Table 1), i.e., among 104 patients unselected for a s u s p i c i o n of constitutional a n o m a l y from t u m o r material analysis. This provides an u p p e r estimate for i n c i d e n c e of constitutional chromosome alterations in our sample: two of 104 = 1.92%, but we obtained information on the constitutional karyotype of some patients retrospectively by analysis of t u m o r material that exhibited either normal karyotypes or karyotypes with a single structural a n o m a l y whose congenital origin could be confirmed by blood l y m p h o c y t e analyses. This increases the total sample to 329 (APC patients included), w h i c h yields the lower estimate of incidence: two of 329 : 0.6% © 1992 Elsevier Science Publishing Co. Inc. 655 Avenue of the Americas~New York. NY 10010
Constitutional Balanced Translocations in Solid Tumors
Table 1
51
Constitutional c h r o m o s o m e anomalies in adult patients with solid tumors
Population
Tissue sample (n)
Non-APC colorectal adenocarcinoma (n = 86)
A(13) B(44)
APC colorectal adenocarcinoma (n : 20) Breast adenocarcinoma (n ~ 107)
C(11] D(18) A(11) B (9) A(16) 9(31)
Cervical carcinoma (n = 91) Meningioma (n = 25)
Constitutional chromosomal aberration fra(10)(q24.2) t(13q14q) fra(2)(q22)
del(5) in two relatives fra(2)(q14.1) -t(13q14q) fra(10)(q25)
C(35) B(25) D(91)
t(3;15)(q13.3;q21)
D(25)
t(3;6)(q13.3;q22)
Abbreviation: APC, adenomatous polyposis coli; n, sample size.
Tissue sample A, 100 lymphocytemetaphases analyzed; tissue sample B, 2-65 lymphocytes metaphases analyzed; tissue sample C, only normal karyotypes observed in tumor biopsy; tissue sample D, normal karyotypes observed beside abnormal karyotypes in tumor biopsy. In addition, for samples C and D, the possible constitutional origin of simple balanced rearrangements was verified by study of blood lymphocytes.
of our cases, w h i c h represents a sixfold increase frequency of Robertsonian translocations as compared with that observed in the n e w b o r n p o p u l a t i o n (p ~ 0.01) (Table 2). The two other balanced rearrangements were reciprocal translocations. Both were detected after analysis of tumor material and retrospective study of lymphocytes. They represent two of 329 -- .6% of studied cases, w h i c h is also sixfold higher than that in the normal population (p 0.01). A m o n g colorectal cancers, APC patients were analyzed separately, because a deletion of 5q arm, in w h i c h the gene is m a p p e d [7, 8] may be a direct etiologic factor [3, 4]. In a sample of 20 patients from 17 i n d e p e n d e n t families, a del(5)(q14q23.1) was observed effectively in two relatives. W h e n APC patients w h o had a special ascertainm e n t are excluded, the total occurrence of constitutional structural rearrangements can be estimated as 1.22%. We cannot draw definite conclusions about this freq u e n c y because of the limited n u m b e r of patients. To obtain
Table 2
a better estimate of i n c i d e n c e of constitutional alterations in individuals with solid tumor, we r e v i e w e d data from the literature. To our knowledge, only two studies refer to significant series of constitutional karyotypes in adult patients affected by solid tumors. In a series of 50 patients with breast cancer, Ochi et al. [9] observed one reciprocal translocation. In another series of 76 cases of breast and 30 cases of lung cancer, Pathak et al. [10] did not report any constitutional c h r o m o s o m a l rearrangement. F r o m these data and ours, we calculated the f r eq u en cy of constitutional c h r o m o s o m e alteration in the p o p u l a t i o n w i t h solid t u m o r and compared it with the i n c i d e n c e at birth (Table 2). Although the sample of patients is still small, it confirms that a significant excess of constitutional balanced rearrangements exists in patients affected by a solid tumor. This excess may be sixfold (Table 2). In corollary, it ma y indicate a higher risk of cancer in i n d i v i d u a l s w i t h such rearrangements, although prospective studies of larger se-
Constitutional balanced c h r o m o s o m e rearrangements observed in the p o p u l a t i o n w i t h solid tumor and in newborns Population with solid tumor
Chromosomal anomaly
Our series (n = 329)
Total c (n = 455)
Robertsonian translocations Reciprocal translocations All translocations Inversions
2 (0.6)a 2 (0.6)° 4 (1.22) ~ 0
2 (0.44) a 3 (0.66) b 5 (1.1) b 0
Newborn population d (n ~ 67,014) 62 60 122 12
(0.09) (0.09) (0.18) (0.02)
n, sample size; percentages are indicated in brackets. Each sample was compared to newborns, and the probability of observingsuch distributions was calculated under a Poisson distribution: ° p ~ 0.01, bp ~ 0.001. ~:Includes our sample and the two published series of Ochi et al. [9] and Pathak et al. [10]. d UNSCEAR[1].
52
ries of patients are necessary before any definite conclusion can be reached. Again because of the small sample size, it would be premature to conclude that specific chromosome breakpoints exist, although chromosome 3 was involved twice in reciprocal translocation after breakage in band 3q13.3.
Constitutional Fragile Sites Our study was obviously not performed u n d e r optimal conditions to detect constitutional fragile sites because a limited n u m b e r of metaphases was studied in most cases. Furthermore, fragile sites are usually expressed in lymphocytes and not in tumor cells [10], and we had information on blood lymphocytes for 130 patients only. In this sample, we detected two cases of fra(10)(q24.2), one case of fra(2)(q14.1), and one case of fra(2)(q22). Another case of fra(10)(q25) was described by Ochi et al. [9] among 50 breast cancer patients; no information was provided by Pathak et al. [10]. This yields a frequency of 2.8% for the detected rare fragile sites, which is in the range of that of the normal population [11, 12], but our observed frequency probably is greatly underestimated and an increased frequency of fragile site carriers among patients with solid tumors cannot be excluded.
REFERENCES 1. UNSCEAR Report (1982): Ionizing radiation: Sources and biological effects. United Nations Scientific Committee on the Effects of Atomic Radiation, United Nations, New York. 2. Sandberg AA (1990): The Chromosomes in Human Cancer and Leukemia, 2nd Ed. Elsevier Science Publishing Co., New York.
F. Richard et al.
3. Herrera L, Kakati S, Gibas L, Pietrzak E, Sandberg AA (1986): Gardner's syndrome in a man with an interstitial deletion of 5q. Am J Med Genet 25:473-476. 4. Hockey KA, Mulcahy MT, Montgomery P, Levitt S (1989): Deletion of chromosome 5q and familial adenomatous polyposis. J Med Genet 26:61-62. 5. Alimena G, Billstr6m R, Casalone R, Gallo E, Mitelman F, Pasquali F (1985): Cytogenetic pattern in leukemic cells of patients with constitutional chromosome anomalies. Cancer Genet Cytogenet 16:207-218. 6. Benitez J, Valcarcel E, Ramos C, Ayuso C, Sanchez Cascos A (1987): Frequency of constitutional chromosome alterations in patients with hematologic neoplasias. Cancer Genet Cytogenet 24:345-354. 7. Bodmer WF, Bailey CJ, Bodmer J, Bussey HJR, Ellis A, Gorman P, Lucibello FC, Murday VA, Rider SH, Scambler P, Sheer D, Solomon E, Spurr NK (1987): Localization of the gene for familial adenomatous polyposis on chromosome 5. Nature 328:614-616. 8. Leppert M, Dobbs M, Scambler P, O'Connell P, Nakamura Y, Stauffer D, Wooward S, Burt R, Hughes J, Gardner E, Lathrop M, Wasmuth J, Lalovel JM, White R (1987): The gene for familial polyposis coli maps to the long arm of chromosome 5. Science 238:1411-1413. 9. Ochi H, Watanabe S, Furuya T, Tsugane S (1988): Chromosome fragility of lymphocytes from breast cancer patients in relation to epidemiologic data. Jpn J Cancer Res 79:1024-1030. 10. Pathak S, Hopwood VL, Hortobagyi GN, Jackson GL, Hughes JI, Melillo D (1991): Chromosome anomalies in human breast cancer: Evidence for specific involvement of lq region in lymphocyte cultures. Anticancer Res 11:1055-1060. 11. Muleris M, Dutrillaux AM, Lombard M, Dutrillaux B (1987): Noninvolvement of a constitutional heritable fragile site at 10q24.2 in rearranged chromosomes from rectal carcinoma cells. Cancer Genet Cytogenet 25:7-13. 12. Takahashi E, Hori T, Murata M (1988): Population cytogenetics of rare fragile sites in Japan. Hum Genet 78:121-126.
