drome of multiple endocrine neoplasia type 1 (MEN-I), 15 patients with sporadic primary hyper- parathyroidism, and SO healthy controls. In both patient groups, ...
Hereditas 108: 227-229 (1988)
Cytogenetical investigations in patients with primary hyperparathyroidism and multiple endocrine neoplasia type 1 LARS BENSON’, KARL-HENRIK GUSTAVSON*, JONAS RASTAD3, GORAN AKERSTROM’, KJELL OBERG‘ and SVERKER LJUNGHALL‘ Departments of Internal Medicine’, Clinical Genetics2,and Surgery3, University Hospital, S-75185 Uppsala, Sweden
s. 1988. Cytogenetical investigations in patients with primary hyperparathyroidism and multiple endocrine neoplasia type 1. -Hereditas 108 227-229. Lund, Sweden. ISSN 0018-0661. Received July 28,1987 BENSON, L., GUSTAVSON, K.-H., RASTAD, J.. AKERSTROM, G., OBERG, K.and LJUNGHALL,
Chromosomal instability can be associated with an increased tendency to develop malignancies. Chromosomal analyses of cultured lymphocytes were performed in 40 members of families with the syndrome of multiple endocrine neoplasia type 1 (MEN-I), 15 patients with sporadic primary hyperparathyroidism, and SO healthy controls. In both patient groups, there were individuals with an increased frequency of chromosomal breakages. The possibility remains that this defect might be related to future development of malignancies. The great overlap between, and within, the MEN-1 families and the others makes chromosomal analyses unsuitable for screening with the aim to detect affected but not yet symptomatic carriers of the MEN-1 gene.
Sverker Ljunghall, Department of Internal Medicine, University Hospital, S-75185 Uppsala, Sweden
The syndrome of multiple endocrine neoplasia type Materials and methods 1 (MEN-1) is an autosomal dominantly inherited disease affecting several endocrine organs (WERMER Patient material 1974), and always involving the parathyroid glands. The study entails 105 individuals. Altogether 26 The parathyroid lesions consist of asymmetrical members from three generations of one family with glandular hyperplasia which generally become clin- MEN-1, aged 5-65 years, were investigated. Six of ically evident prior to the others, sometimes preced- the children were younger than 10years and 13were ing them with decades (BALLARDet al. 1964;JOHNSON10-20 years old. In another seven families, 14 paet al. 1967; MARXet al. 1977; LAMERSand FROELINGtients with MEN-l were also studied. The criteria 1979; BENSONet d.1987). for MEN-1 were identical to these previously pubIn a previous, preliminary, report an increased lished (OBERG et al. 1982; MUHRet al. 1984). In adfrequency of chromosome breakages was found in dition, 15 consecutive patients with sporadic, pripatients with MEN-1 (GusTAvsoNet al. 1983). Since mary HPT (10 women, 5 men, age 35-84 years) the MEN-1 syndrome is potentially fatal, it is urgent were investigated preoperatively. As controls, 50 to diagnose the trait as early as possible. In the hope apparently healthy individuals between 6 and 70 that cytogenetical investigations might provide a ge- years of age were studied. netic “marker”, we have performed the present, more extensive, study of affected and healthy mem- Methods bers of MEN-1 families. Patients with sporadic (i.e., non-familial) primary hyperparathyroidism (HPT) Total serum calcium (reference range 2.20-2.60 were also included, as an association between pri- mmol/l) concentrations were determined by atomic mary HPT and other tumours of both endocrine and absorption and adjusted to the prevailing albumin non-endocrine origin has been reported (WAJNGOTconcentration as previously described, while serum et al. 1980; P A L M ~al.R1987). ~ ~ Thus, both in the parathyroid hormone (PTH) concentrations were familial and sporadic forms of HPT there could be determined by a radioimmunoassay method (BENan inherited predisposition involved, possibly SON et al. 1987). Chromosome analyses were performed on cells caused by cytogenetic instability.
228
Hereditas 108 (1988)
L. BENSON ET AL.
Table I . Chromosome breakages in MEN-I families
Sex
M M M M M M M F
M F F F F M
F F F F
M F F M F F
M M
M F F F F
M F
M F F
M F F F
Age Subject
65 39 18 16 12 34 37 28 61 34 I1 6 37 10 38 31 1 18 55 21 21 9 31 12 9 5 53 57
64 62 60 33 47 51 51 23 21 25 19 64
AA AAA AAAA AAAB AAAC AAB AAC AAD AB ABA ABAA ABAB ABB ABBA ABC ABD ABDA ABE AC ACA ACB ACBA ACC ACCA ACCB ACCC BCA CA CB CC DA DAA F J LA LAA LAB LAC LAD PA
At risk 1 1 1
1 1 1
1 1 1 1 1 1 1 1 1 1 2 1 1
Total breaks/ Verified lesion 100cells 4 4 1 8 17 8 6 14 11 0 3 6 2
1
1
2 4 1 12 5 8 7 16 12 7 23 11 0
1
4
I 1 1
4 0 6 2 0 6 0 5 18 4 12 2
1
1 1 1 2 2 2
1 1 1
1 1 1 1
1 1
HPT+EPT HPT+EPT 0 0
0 HPT 0 0 HPT+EFT HPT+EPT 0 0
HPT+EPT 0 0
0 0 HPT HFT+EPT H E + EPT+PT HPT+ EFT 0 0 0
0 0
HPT+EPT HPT+PT HPT+EPT+PT 0 HPT+EPT HPT+EPT+PT HPT+EPT+PT HPT+PT HPT HPT 0 0
0 HPT+EPT+PT
The family is denoted by the first letter, childrenby the following letter (AAA isthechildof AA,whoisachildofA) At risk(1)denotesachildofanaffected parent, (2) of a healthy parent HFT = hyperparathyroidism EPT = endocrine pancreatic turnour PT = pituitary turnour
from phytohaemagglutinin-stimulated lymphocyte cultures. The culture medium was 8 ml of Parker 199 (Flow Labs) with 2 ml of human serum (Flow Labs) supplemented with penicillin, streptomycin and phytohaemagglutinin. The cultures were incubated for 72 h at 37°C. Sixteen hours before the end of the culturing time, Colcemid 0.1 pglml was added. Conventional air-dried slides were prepared and Giemsa stained for analyses of chromosome aberrations. The aberrations were recorded in accordance with the WHO recommendations (BUCKTON and EVANS 1973; NORDENSON 1979). The
Table 2. Chromosome (chromatid normal controls
Control groups
+ isochromatid) breakages in
Number of individuals
6-15 years of age 23-50 years of age 51-70yearsof age
1 29 14
Chromosome breaks per 100 cells Mean t SD
Range
4.5k2.0 3.4k1.8 6 f2.2
2- 6 0-14 2-15
Table 3. Chromosome breakages in patients with sporadic hyperparathyroidism
Sex
Age
Sca mmolil PTH arbUil (2.2Lk2.60)” (0.4&1.08)”
Total breaks/ l00cells
F M
72 35
F
55
10 0 26
F F M F
84 84 73 60 81 66 61 52 59 65 19 67
3.25 2.19 3.22 2.16 3.07 2.71 2.80 3.55 2.97 2.87 2.83 2.94 3.22 3.05 3.77
F M F
M M
F F F a
2.27 0.99 1.71 2.12 2.85 0.72 1.10 2.0 1.87 1.05 0.70 0.99 0.86 0.91 1.60
1 0
9 0 13 0 13 1 11 15 2 0
Reference range
numbers of chromosome (chromatid and isochromatid) breaks in 100 cells from each person were analysed.
Results Table 1summarizes results of chromosome analyses of the 40 members of MEN-1 families, and the number of chromosome breaks in the control samples from healthy individuals are given in Table 2. There was a large variation in the number of chromosome breakages among the patients. Although no or only a few breakages were seen in the majority of the investigated individuals, some of them had as many as 20 % of all cells affected. However, there was no consistent pattern and there was no statistically significant difference in the number of chromosome breakages between affected and non-affected members of the MEN-1 families or between affected members and healthy controls. Nor was there any difference in the number of chromosome breakages between those who had a more advanced MEN-1 syndrome compared to those who
Hereditas 108 (1988)
only had HPT. Among the patients with sporadic, non-familial HPT (Table 3), there was also a wide range of chromosomal findings but there was no relationship between the number of chromosome breakages and either serum calcium or PTH concentrations. In general, the cytogeneticai results in this group were similar to those among the MEN-1 families.
Discussion The present study disclosed no consistent pattern of chromosomal instability in family members with the MEN-1 syndrome. However, several individuals in the affected families clearly had an increased number of chromosome breakages compared to healthy controls. This, however, did not appear to be a specific trait exclusive for the carrier of the MEN-1 trait. Therefore, analysis of chromosome breakages does not appear to be specific enough to reveal early cases of MEN-1, before HPT can be detected biochemically. The possibility remains, however, that chromosome breakages could reflect a predisposition for a more aggressive disease, e.g., earlier development of malignant pancreatic lesions. Thus, non-affected members from some, but not all, MEN-2 families do show enhanced sensitivity to chromosomal damaging effects of bleomycine (TSIONPRA 1986), which suggests a “variable” degree of genetic heterogenity among MEN-2 families. SAMAAN et al. (1984) also reported a high rate of chromosome aberrations in cultured lymphocytes of three families with MEN-2a and in patients with medullary carcinoma of the thyroid. The finding that in some patients with sporadic HPT there was also a clearly increased number of chromosomal breakages might be considered as reflecting a similar propensity since parathyroid hormone ( P E R R i s e t al. 1971) and calcium ( P E R R i s e t al. 1968; LUCKASEN et al. 1974) have mitogenic properties. At present, there is no evidence among the investigated patients of any other tumours, but we have recently shown (PALMER et al. 1988) that patients with primary HPT appear to run an increased risk of developing malignant disorders for many years subsequent to parathyroid surgery.
MULTIPLEENDOCRINE NEOPLASIATYPE I
229
Literature cited J . 1964. Familial multiple endocrine adenoma-peptid ulcer complex. -Medicine 42: 44814516 BENSON, L., LIUNGHALL, S . , AKERSTROM, G. and OBERG,K. 1987. Hyperparathyroidism presenting as the first lesion in multiple endocrine neoplasia type l. - A m . J. Med. 82: 731-737 BUCKTON,K . E. and EVANS, H. J. 1973. Methodsfor the analysis of human chromosomal abberations. - WHO, Geneva GUSTAVSON, K. H., JANSSON, R. and C)BERG, K. 1983. Chromosomal breakage in multiple endocrine adenomatosis (types I and 11). - Clin. Genet. 23: 143-149 JOHNSON, G. J . , SUMMERSKILL, W . H., ANDERSON, V. E. and KEATING, F. R. 1967. Clinical and genetic investigation of a large kindred with multiple endocrine adenomatosis. - New Engl. J . Med. 277: 137%1385 LAMERS, C. B . H. W . and FROELING, P. G . A . M. 1979. Clinical significance of hyperparathyroidism in familiar multiple adenomatous type 1 (MEA-1). - A m . J . Med. 66: 422-424 LUCKASEN, J. R., SHITE, K. G. and KERSEY, J. H. 1974. Mitogenic properties of a calcium ionophore, A2318. - Proc. Natl. Acad. Sci. 71: 5088-5090 MARX, S . J . , SPEGEL, A. M., BROWN, E. M. and AUERBACH, G. D. 1977. Family studies in patients with primary parathyroid hyperplasia. - A m . J. Med. 62: 69&-705 MUHR, C., LJUNGHALL, S . , AKERSTROM, G. et al. 1984. Screening for multiple endocrine neoplasia syndrome (type I ) in patients with primary hyperparathyroidism. - Clin. Endocrinol. 20: 153-162 NORDENSON, I . 1979. Clinical and experimental studies of chromosomal aberrations in cultured leukocytes. - Umed University Medical Dissertations. New series no. 43 OBERG, K . , WALINDER, BOSTRCSM, H . , LUNDKVIST. G. and WIDE, L. 1982. Peptide hormone markers in the screening for endocrine tumors in membersof MEA-1 families. - A m . J. Med. 73:619-630 PALMCR, M., ADAMI, H.-O., KRUSEMO, U. B. and LIUNGHALL, S. 1988. Increased risk of malignant diseases after operation for primary hyperparathyroidism: a nationwide cohort study. - A m . J. Epidemiol. (accepted) PERRIS, A . D.. WHITFIELD, J. F. and TOLG, P. K . 1968. Role of calcium in the control of growth and cell division. -Nuture219: 527-529 PERRIS,A . D., MACMANUS, J. P., WHITFIELD, J . F.and WEISS, L. A. 1971. Parathyroid gland and mitotic stimulation in the rat bone marrow after hemorrhage. - A m . J. Physiol. 220: 773-778 SAMAAN, N . A . , HSU, T. C., PATHAK, S., SAAD, M . F., ORDONEZ, N . G. and HICKEY,R. C. 1984. Chromosomal instability in medullary carcinoma of the thyroid. - World 1. Surg. 8: 487-492 TSIONPRA, K. 1986. Abstr 7th Int. Congr. Human Genetics, Berlin WAJNGOT, A , , WERNER, S., GRANBERG, P.-0.. LINDVALL, N . 1980. Occurrence of pituitary adenomas and other neoplastic diseases in primary hyperparathyroidism. - Surg. Gynecol. Obstet. 1 S l : 401403 WERMER, P. 1974. Multiple endocrine adenomatosis; multiple hormone-producing tumors a familial syndrome. - Clin. Gastroenterol. 3 : 671484 BALLARD, H., FRAME, B . andHARTSOCK, R.
o.,
