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Chromosomal differences between acute nonlymphocytic leukemia in patients with prior solid tumors and prior hematologic malignancies
Chromosomal Differences Between Acute Nonlymphocytic Leukemia in Patients with Prior Solid Tumors and Prior Hematologic Malignancies A Study of 14 Cases with Prior Breast Cancer Zissis Mamuris, Jeanine Dumont, Bernard Dutrillaux, and Alain Aurias
ABSTRACT: A cytogenetic study of 14 patients with secondary acute nonlymphocytic leukemia (SANLL) with prior treatment for breast cancer is reported. The chromosomes recurrently involved in numerical or structural anomalies are chromosomes 7, 5, 17, and 11, in decreasing order of frequency. The distribution of the anomalies detected in this sample of patients is similar to that observed in published cases with prior breast or other solid tumors, though anomalies of chromosome 11 were not pointed out, but it significantly differs from that of the SANLL with prior hematologic malignancies. This difference is principally due to a higher involvement of chromosome 7 in patients with prior hematologic malignancies and of chromosomes 11 and 17 in patients with prior solid tumors. A genetic determinism involving abnormal recessive alleles located on chromosomes 5, 7, 11, and 17 uncovered by deletions of the normal homologs may be a cause of S-ANLL. The difference between patients with prior hematologic malignancies or solid tumors may be explained by different constitutional mutations of recessive genes in the two groups of patients. INTRODUCTION In a recent survey of the literature, Bloomfield [1] analyzed the occurrence of anomalies of chromosome 5 and 7 in acute nonlymphocytic leukemia (S-ANLL) secondary to prior m a l i g n a n c y following treatment with chemo- or radiotherapy. It was concluded that specific rearrangements of chromosome 5 (principally del(5q)) correlated with prior solid tumors, whereas loss of whole chromosome 5 and/or 7 and gains of other chromosomes correlated with prior hematologic malignancies. To date, cytogenetic information for ANLL secondary to prior solid tumors rem a i n s limited, with only a few published series [2-11]. We have studied a series of
From the C. N. R. S. URA 620 Structure et Mutagen~seChromosomiques, Institut Curie, Section de Biologie (Z. M., B. D., A. A.) and SectionM6dicale (J. D.), Paris, France.
Address reprint requests to: Aiain Aurias, C. N. Ft. S. URA 620 Structure et Mutagen~se Chromosomiques, Institut Curie, Section de Biologie, 26 rue d'Ulm, 75231 Paris, Cedex 5, France. Received December 29, 1988; accepted March 31, 1989.
43 O 1989 ElsevierSciencePublishingCo., Inc. 655 Avenue of the Americas, New York, NY 10010
Cancer Genet Cytogenet42:43-50 (1989) 0165-4608/89/$03.50
44
z. Mamuris et al. 14 cases of S-ANLL following breast cancer and compared their chromosomal anomalies to those of S-ANLL following various other malignancies. We confirmed that the chromosomal patterns of S-ANLL following solid tumors and hematologic malignancies differ, though our conclusions differ from those of Bloomfield [1].
MATERIALS AND METHODS Bone marrow and blood samples were obtained from 14 patients with S-ANLL with prior treatment for breast cancer. Cultures were carried out in TC199 (6.5 volume) and h u m a n serum (1.5 volume) for 24 and 48 hours. Colchicine block, spreading, and R banding were performed according to usual methods [12]. All the rearrangements observed were classified according to ISCN [13]. The numbers of analyzed mitoses are indicated for each case in Table 1.
RESULTS Karyotypic formulas of the 14 patients are given in Table 1. Among the 35 anomalies detected, there are 14 derivative chromosomes from unbalanced rearrangements leading to deficiencies, seven reciprocal translocations, four trisomies, four monosomies, two deletions, two ring chromosomes, and two unidentified markers. All these anomalies are distributed among the leukemic cells from 12 patients, a normal karyotype being observed in the two others. Structural rearrangements account for 27 of 35 (77%) of all anomalies (Table 1). The chromosomes most frequently affected are: 7 (six times); 5, 8, and 17 (five times); 11 (four times); and 1, 6, 12, and 22 (two times). Among these anomalies, chromosome 7 is totally missing in three cases and chromosome 8 is in excess in two cases. The most frequent deficiencies, resulting from both rearrangements and whole chromosome losses, are: 17p (five times); 7q (four times); and 5q and 7p (three times). Chromosome 11 is the most frequently involved in reciprocal translocations (three times), but without recurrent breakpoints. However, a t(9;11)(q21;q24) observed in one of our patients (case 13) has been reported to be recurrent in both secondary and de novo ANLL [1, 14, 15].
Comparison of Our Results with Published Data For this comparison we selected sizeable series with description of abnormal chromosomes [2, 3, 6-9] and created three categories including 19 cases with prior breast cancer, 46 cases with prior other solid tumors, and 89 cases with prior malignant hemopathies (principally lymphomas). For the comparison with our data, we took into consideration all anomalies of chromosomes 3, 5, 6, 7, 11, 12, and 17 (Table 2; Fig. 1) because attention was drawn to them by various authors or they appeared to be frequently affected in our study. The results on the distribution of the anomalies of these chromosomes are as follows: The two samples of "breast cancer" (literature and our own) are very similar) (x 2 = 0.46, v = 3, nonsignificant), The sum of the two "breast cancer" samples is very similar to the sample "other solid tumors" (×2 = 1.69, v = 4), The sum of all solid tumors is significantly different from the "malignant hemopathies" sample (x 2 = 19.4, v = 6, p ~ 0.005). This difference is principally due to an overinvolvement of chromosome 7 in the "malignant hemopathies" sample (x 2 participation = 5.1, v = 1) and of chromosomes 11 and 17 in the "solid tumors"
+ -
-
5. 6.
7. 8. 9.
+ + +
+ + -
+ +
+ + +
+
+
+ + +
Radiotherapy
34 84 60
168 48
120 45 93
90 75
44
23 73 51
Time primary disease/S-ANLL
+ + -
+
+ + -
+ +
+
+ +
Preleukemia
LAM4 LAM4 ?
LAM2 LAM6
LAM2 LAM2 LAM2
LAM2 LAM2
LAM2
LAM5A ? LAM4
0/5 2/17 ?/O
6/0 0/33
24/46 0/18 0/24
0/29
7/10
0/19
0/10 0/41 0/6
No. normal/ FAB abnormal classification mitoses
(q13.2;qll.1),+r(?) Normal karyotype 53,X,-X,+2,+6,del(17)(p11),+der(1)t(1;8) (q24;q21), + der(1)t(1;8)(q24;q21), +der(8)t(1;8)(q24;21),+ der(5)t(5;14) (p14;q21),+ d e r ( 7 ) , + der(20) 46,XX,- 7,del(5)(p14),+mar 46,XX,t(9;11)(q21;q24) Normal karyotype
44,XX,-5,-7,-16,-17,+der(5)t(5;17)
46,XX,t(lO;11)(p12;q14} 4 6 , X X , - 5 , - 6 , - 17,t(4;15)(q24;q26), t ( 1 I ; 2 1 ) ( p 1 2 . 2 ; p 1 2 ) , + der(5)t(5;17) (q21;q22), + der(6)t(6;?)(q22.1;?),+ m a r 47,XX,+ 8,t(7;12)(q11.2;pter) 47,XX,÷8
+der(22)t(22;?),+ der(22)t(22;?)
(p12 ;ql 1.2), + der(5)t(5;17)(qll ;ql 1),
44,XX,-3,-5,-17,-17,-22,-22,r(11), t(12;13)(p12.1;q13),+der(3)t(3;17)
4 6 , X X , - 7,+der(7)t(7;?}{q22;?} 46,XX,t(8;18)(p21.2;q21.2) 45,XX,-7
Karyotypic formulas
f o r m u l a s f r o m t h e 14 p a t i e n t s w i t h p r i o r b r e a s t c a n c e r ~
° All patients except cases 6 - 8 received a chemotherapy treatment including an alkylating agent (melphalan and/or cyclophosphamide). All radiotherapy treatments were localized to breast area at a total dose of 55 Gy, fractionated in 1.8-Gy courses.
Abbreviations: Ad, adriamycin; Am, ametycin; Bleo, bleomycin; 5-FU, 5-fluorouracil; Eto, etoposide; Met, methotrexate; Ten, teniposide; Vin, vincristine.
12. 13. 14.
10. 11.
Ad, 5FU, V i n Ten, Mef, Bleo Ad, 5FU, T e n , M e t +
Ad, 5FU Ad, 5FU, Eto, M e t Ad, A m , 5 F U Vin, M e t 5FU, M e t
1. 2. 3.
4.
Chemotherapy
Clinical data and karyotypic
Case
Table 1
46
Z. Mamuris et al.
60
50
40
30
20
10
3
5
6
7
ii
12
17
Figure 1 Histograms indicating the participation (in percentage) of chromosomes 3, 5, 6, 7, 11, 12 and/or 17 in the various groups of S-ANLL with prior hematologic malignancy (white), breast cancer, from published data (dark grey) and the present study (black), other solid tumors (light grey), and all solid tumors (hatched).
sample (x 2 participation = 6.5 and 5.2, ~ = 1 for chromosomes 11 and 17, respectively). A d d i t i o n a l information is given by the analysis of the types of rearrangements. Rearrangements affecting the four most frequently i n v o l v e d c h r o m o s o m e s are indicated in Table 3 in relation to the prior malignancy. As previously noted, the complete loss of c h r o m o s o m e 7 is more frequent in S-ANLL with prior hematologic
Table 2
Cases in w h i c h an anomaly of ch r o m o so m es 3, 5, 6, 7, 11, 12 and/or 17 were described
Chromosome
Prior hematologic malignancies
Prior breast cancer
Prior breast cancer (present study)
Prior other solid tumors
Prior solid tumors
3 5 6 7 11 12 17 Sample sizes
9 37 18 60 8 13 14 89
2 8 2 6 3 2 6 19
1 5 2 6 4 1 4 14
6 17 4 17 12 9 15 48
9 30 8 29 19 12 25 81
Source: References [2, 3, 6-8] and present study.
47
Chromosomal Differences in ANLL Table 3
The most frequent chromosome anomalies in S-ANLLa
Anomalies
Prior solid tumors
Prior hematologic malignancies
del(5q) --5 del(7q) -7 del(11) -11 trcp(11;other) del(17p) -17 trcp(17;other) Whole sample size
18 7 8 17 7 0 11 13 6 4 81
17 10 13 41 4 2 0 8 2 2 89
a Unbalanced rearrangements resulting in a deficiencyof the correspondingchromosome are grouped with deletions.
malignancies than with prior solid tumors (x 2 = 16.3, v = 1). Very striking is the high rate of reciprocal translocations of chromosome 11 in S-ANLL with prior solid tumors and its absence in S-ANLL with prior hematologic malignancies. The rates of rearrangements and/or losses of chromosome 5 are very similar in the two samples. DISCUSSION The value of the study of S-ANLL has been stressed by several authors because this leukemia represents a quasiexperimental model for studying a leukemogenic process. Characteristic recurrent anomalies of chromosomes 5 and 7 and, to a lesser degree, of chromosome 17, have already been described [2-11]. The extensive study of Bloomfield [1] concluded that the distribution of anomalies of chromosomes 5 and 7 may vary in relation to the prior malignancy: del(5q) may be characteristic of cases with prior solid tumors, whereas losses of chromosomes 5 and 7, and gains of others chromosomes, would seem to characterize cases with prior hemopathies. Our data on S-ANLL with prior breast cancer are not in contradiction with this proposal: three of three anomalies of chromosome 5q led to deletions (Table 1). However, we also found three cases with loss of chromosome 7, indicating that if the distinction is valid, it is statistical rather than absolute. Additional information on differences between the two samples were found in our study. Chromosomes 5 and 7 were the most frequently affected (in 37% and 34%, respectively, of cases with prior solid tumors). The higher rate of complete loss of chromosome 7 in the "hematologic" versus the "solid tumor" group was confirmed. However, we could not confirm the difference in chromosome 5 involvement as proposed by Bloomfield [1] because this chromosome is equally frequently missing or deleted in both groups of S-ANLL. In the solid tumor group, anomalies of chromosomes 17 and 11 are almost as frequent (in about 29% and 26% of cases, respectively) as those of chromosomes 5 and 7. The frequent involvement of chromosome 17 was previously noticed in S-ANLL with prior solid tumors [7]. Chromosome 11 is principally involved in reciprocal translocations, but no recurrent breakpoints could be found. This excess of lesions of chromosomes 11 and 17 was not observed in S-ANLL with prior malignant hemopathies because they were affected in about 16% and 9% of the cases, respectively, which is not more than several other chromosomes (Fig. 1). It would be of great interest to understand the causes of such differences. A n excess of aberrations affecting chromosome 11 has
48
Z. Mamuris et al. been described after treatment by etoposide and cisplatin [16]. Three of our patients received etoposide or teniposide (cases 2, 9, and 10}, but none of them exhibited any anomaly of chromosome 11 in their leukemic clone. Nevertheless, it remains possible that the type of chromosomal anomalies largely depends on the treatment received for the first malignant disease. Indeed, these treatments differ in the groups with prior solid tumors and malignant hemopathies. The fact that in vitro and in vivo treatments by melphalan {an alkylating agent) induce a high rate of anomalies of chromosomes 11 and 17 [17, 18] and that alkylating agents are quite frequently used in treatment of solid tumors (11 of 14 of our patients; Table 1} may be of importance. However, such drugs are also used in the treatment of hematologic malignancies, and the above explanation may be insufficient for discriminating between the two groups. It would be of great interest to study the respective role of radiation and of the various drugs used for chemotherapy. Unfortunately, the therapeutic protocols vary, and such a comparative study would be premature at the present time. Another element of importance may be the genetic constitution of the patients. In a variety of solid tumors, such as colorectal and breast cancers, rhabdomyosarcomas, and hepato- and nephroblastomas, recurrent deletions and/or allelic losses of chromosomes 11 and 17 have been described [19-25]. This suggests the existence of recessive genes located on these chromosomes, which might be involved in one step of the tumoral process. As indicated by familial studies with recurrent cancers, the abnormal recessive alleles may be transmitted as predisposing factors. Thus, they may exist constitutionally in single copy in a number of individuals. In S-ANLL patients, such genetic determinism can be suspected for recessive genes located on chromosomes 5, 7, 11, and 17, which would be uncovered by deletions of their normal homologs, as we proposed earlier [18]. Thus, it is possible that, among patients with prior solid tumors, several carried abnormal recessive alleles on chromosomes 17p and 11, which might account for the higher involvement of these chromosomes in this group than in that with prior hematologic malignancies. The presence of preexisting mutations may also provide some explanation for the occurrence of SANLL in patients with prior solid tumors treated by surgery alone, as described by Zaccaria et al. [10]. However, this explanation may be insufficient for chromosome 11 because this chromosome is involved in various reciprocal translocations without any visible deletions. The variety of the breakpoints excludes the involvement of a single gene, but it is possible that several genes of this chromosome play a role in tumorigenesis, as suggested by the fact that deletions of the short and long arms are observed in some tumors, such as breast cancer [20]. Another mechanism, recently described, may be also pertinent here: it has been shown that several genes of chromosome 11 are abnormally methylated in a variety of tumor cell lines [26]. If this methylation pattern is determined by a single gene, as seems to be the case for the late replicating X, a relationship may exist between mutagenesis and a multilocus inactivation. This work was supported by grants B16-147F from EURATOM, the Ligue Nationale Fran~aise Contre le Cancer, and the Minist~re de la Recherche et de l'Enseignement Supbrieur.
REFERENCES
1. Bloomfield CD (1986): Chromosome abnormalities in secondary myelodysplastic syndromes. Scand ] Haematol 36 {Suppl 45):82-90. 2. Rowley ]D, Golomb HM, Vardiman ] (19771: Nonrandom chromosomal abnormalities in acute nonlymphocytic leukemia in patients treated for Hodgkin disease and non-Hodgkin lymphomas. Blood 50:759-770.
Chromosomal Differences i n ANLL
49
3. Rowley JD, Golomb HM, Vardiman JW (1981): Non-random chromosome abnormalities in acute leukemia and dysmyelopoietic syndromes in patients with previously treated malignant disease. Blood 58:759-767. 4. Sandberg AA, Abe S, Kowalczyk JR, Zedgenidze A, Takeuchi J, Kakati S (1982}: Chromosomes and causation of human cancer leukemia. L. Cytogenetics of leukemias complicating other diseases. Cancer Genet Cytogenet 7:95-136. 5. Pedersen-Bjergaard J, Philip P, Mortenson RT, Ersboll J, Jansan G, Panduro J, Thomsen M {1981): Acute nonlymphocytic leukemia, preleukemia, and acute myeloproliferative syndrome secondary to treatment of other malignant diseases. Clinical and cytogenetic characteristics and results of in vitro culture of bone marrow and HLA typing. Blood 57:712-723. 6. Pedersen-Bjergaard J, Philip P, Pedersen NT, Hou-Jensen K, Svejgaard A, Jensen G, Nissen NI (1984): II. Bone marrow cytology, cytogenetics, results of HLA typing, response to antileukemic chemotherapy and survival in a total series of 55 patients. Cancer 54:452-462. 7. Groupe Fran~ais de Cytog~n~tique H~matologique (1964): Chromosome analysis of 63 cases of secondary nonlymphoid blood disorders: A cooperative study. Cancer Genet Cytogenet 12:95-104. 8. Arthur DC, Bloomfield CD (1984): Banded chromosome analysis in patients with treatment-associated acute nonlymphocytic leukemia. Cancer Genet Cytogenet 12:189-199. 9. Le Beau MM, Albain KS, Larson RA, Vardiman JW, Davis EM, Blough RR, Golomb HM Rowley JD (1986): Clinical and cytogenetic correlations in 63 patients with therapy-related myelodysplastic syndrome and acute nonlymphocytic leukemia: Further evidence for characteristic abnormalities of chromosomes No. 5 and 7. J Clin Oncol 4:325-345. 10. Zaccaria A, Alimena G, Baccarani M, Billstr6m R, Carbonell F, Castoldi GL, Fuscaldo K, Hecht F, Hossfeld DK, Mitelman F, Rosti G, Sandberg AA, Tascinari A, Testoni N, Tura S (1987): Cytogenetic analyses in 89 patients with secondary hematologic disorders--results of a cooperative study. Cancer Genet Cytogenet 26:65-74. 11. Third MIC Cooperative Study Group (19871: Morphologic, Immunologic and Cytogenetic CMIC) working classification of the primary myelodysplastic syndromes and therapy-related meylodysplasias and leukemias. Cancer Genet Cytogenet 32:1-10. 12. Dutrillaux B, Couturier J (1981): La Pratique de l'Analyse Chromosomique. Masson, Paris. 13. ISCN (1985): An International System for Human Cytogenetic Nomenclature, Harnden DG, Klinger HP (eds.); published in collaboration with Cytogenet Cell Genet (Karger, Basel, 19851; also in Birth Defects: Original Article Series, VoL 21, No. 1 (March of Dimes Birth Defects Foundation, New York, 1985). 14. Hagemeijer A, H~ihlem K, Sizoo W, Abels J (1982): Translocation (9;11}(p21;q23) in three cases of acute monoblastic leukemia. Cancer Genet Cytogenet 5:95-105. 15. Fourth International Workshop on Chromosomes in Leukemia, 1982 (1984): Rearrangements of llq. Cancer Genet Cytogenet 11:294-295. 16. Ratain MJ, Kaminer LS, Bitran JD, Larson RA, Le Beau MM, Skosey C, Purl S, Hoffman PC, Wade J, Vardiman JW, Daly K, Rowley JD, Golomb HM C1987}: Acute nonlymphocytic leukemia following etoposide and cisplatin combination chemotherapy for advanced nonsmall-cell carcinoma of the lung. Blood 70:1412-1417. 17. Mamuris Z, Prieur M, Dutrillaux B, Aurias A (1989): The chemotherapeutic d~ug melphalan induces breakage of chromosomes regions rearranged in secondary leukemia. Cancer Genet Cytogenet 37:65-77. 18. Mamuris Z, Gerbault-Seureau M, Prieur M, Pouillart P, Dutrillaux B, Aurias A (1989): Chromosomal aberrations in lymphocytes of patients treated by melphalan. Int J Cancer 43:80-66. 19. Koufos A, Hansen MF, Copeland NG, Jenkins NA, Lampkin BC, Cavenee WK (!985): Loss of heterozygosity in three embryonal tumors suggests a common pathogenetic mechanism. Nature 316:330-334. 20. Das DD, Schroeder WT, Chao LY, Kikuchi H, Strong LC, Riccardi VM, Pathak S, Nichols W-W, Lewis WH, Saunders GF (1978): Genetic mechanisms of tumor-specific loss of 11p DNA sequences in Wilms' tumor. Am J Hum Genet 41:202-217. 21. Gerbault-Seureau M, Vielh P, Zafrani B, Salmon R, Dutrillaux B (1967): Cytogenetic study of twelve human near-diploid breast cancers with chromosomal changes. Ann Genet 30:138-145. 22. Ali IV, Lidereau R, Theillet C, Callahan R (1987): Reduction to homozygosity of genes on chromosome 11 in human breast neoplasia. Science 238:185-186.
50
Z. M a m u r i s et al.
23. Monpezat JP, Delattre O, Bernard A, Grunwald D, Remvikos Y, Muleris M, Salmon R], Frelat G, Dutrillaux B, Thomas G (1988): Loss of alleles on chromosome 18 and on the short arm of chromosome 17 in polyploid colorectal carcinomas. Int J Cancer 41:404-408. 24. Vogelstein B, Fearon ER, Hamilton SR, Kern SE, Preisinger AC, Leppert M, Nakamura Y, White R, Smits AM, Bos JL (1988): Genetic alterations during colorectal tumor develop,ment. N Engl J Med 319:525-532. 25. Muleris M, Salmon RJ, Zafrani B, Girodet J, Dutrillaux B (1985): Consistent deficiencies of chromosome 18 and of the short arm of chromosome 17 in eleven cases of human large bowel cancer: a possible recessive determinism. Ann Genet 28:206-213. 26. Bustros A de, Nelkin BD, Silverman A, Ehrlich G, Poiesz B, Baylin SB (1988): The short arm of chromosome 11 is a "hot spot" for hypermethylation in human neoplasia. Proc Natl Acad Sci USA 85:5693-5697.
ABSTRACT: A cytogenetic study of 14 patients with secondary acute nonlymphocytic leukemia (SANLL) with prior treatment for breast cancer is reported. The chromosomes recurrently involved in numerical or structural anomalies are chromosomes 7, 5, 17, and 11, in decreasing order of frequency. The distribution of the anomalies detected in this sample of patients is similar to that observed in published cases with prior breast or other solid tumors, though anomalies of chromosome 11 were not pointed out, but it significantly differs from that of the SANLL with prior hematologic malignancies. This difference is principally due to a higher involvement of chromosome 7 in patients with prior hematologic malignancies and of chromosomes 11 and 17 in patients with prior solid tumors. A genetic determinism involving abnormal recessive alleles located on chromosomes 5, 7, 11, and 17 uncovered by deletions of the normal homologs may be a cause of S-ANLL. The difference between patients with prior hematologic malignancies or solid tumors may be explained by different constitutional mutations of recessive genes in the two groups of patients. INTRODUCTION In a recent survey of the literature, Bloomfield [1] analyzed the occurrence of anomalies of chromosome 5 and 7 in acute nonlymphocytic leukemia (S-ANLL) secondary to prior m a l i g n a n c y following treatment with chemo- or radiotherapy. It was concluded that specific rearrangements of chromosome 5 (principally del(5q)) correlated with prior solid tumors, whereas loss of whole chromosome 5 and/or 7 and gains of other chromosomes correlated with prior hematologic malignancies. To date, cytogenetic information for ANLL secondary to prior solid tumors rem a i n s limited, with only a few published series [2-11]. We have studied a series of
From the C. N. R. S. URA 620 Structure et Mutagen~seChromosomiques, Institut Curie, Section de Biologie (Z. M., B. D., A. A.) and SectionM6dicale (J. D.), Paris, France.
Address reprint requests to: Aiain Aurias, C. N. Ft. S. URA 620 Structure et Mutagen~se Chromosomiques, Institut Curie, Section de Biologie, 26 rue d'Ulm, 75231 Paris, Cedex 5, France. Received December 29, 1988; accepted March 31, 1989.
43 O 1989 ElsevierSciencePublishingCo., Inc. 655 Avenue of the Americas, New York, NY 10010
Cancer Genet Cytogenet42:43-50 (1989) 0165-4608/89/$03.50
44
z. Mamuris et al. 14 cases of S-ANLL following breast cancer and compared their chromosomal anomalies to those of S-ANLL following various other malignancies. We confirmed that the chromosomal patterns of S-ANLL following solid tumors and hematologic malignancies differ, though our conclusions differ from those of Bloomfield [1].
MATERIALS AND METHODS Bone marrow and blood samples were obtained from 14 patients with S-ANLL with prior treatment for breast cancer. Cultures were carried out in TC199 (6.5 volume) and h u m a n serum (1.5 volume) for 24 and 48 hours. Colchicine block, spreading, and R banding were performed according to usual methods [12]. All the rearrangements observed were classified according to ISCN [13]. The numbers of analyzed mitoses are indicated for each case in Table 1.
RESULTS Karyotypic formulas of the 14 patients are given in Table 1. Among the 35 anomalies detected, there are 14 derivative chromosomes from unbalanced rearrangements leading to deficiencies, seven reciprocal translocations, four trisomies, four monosomies, two deletions, two ring chromosomes, and two unidentified markers. All these anomalies are distributed among the leukemic cells from 12 patients, a normal karyotype being observed in the two others. Structural rearrangements account for 27 of 35 (77%) of all anomalies (Table 1). The chromosomes most frequently affected are: 7 (six times); 5, 8, and 17 (five times); 11 (four times); and 1, 6, 12, and 22 (two times). Among these anomalies, chromosome 7 is totally missing in three cases and chromosome 8 is in excess in two cases. The most frequent deficiencies, resulting from both rearrangements and whole chromosome losses, are: 17p (five times); 7q (four times); and 5q and 7p (three times). Chromosome 11 is the most frequently involved in reciprocal translocations (three times), but without recurrent breakpoints. However, a t(9;11)(q21;q24) observed in one of our patients (case 13) has been reported to be recurrent in both secondary and de novo ANLL [1, 14, 15].
Comparison of Our Results with Published Data For this comparison we selected sizeable series with description of abnormal chromosomes [2, 3, 6-9] and created three categories including 19 cases with prior breast cancer, 46 cases with prior other solid tumors, and 89 cases with prior malignant hemopathies (principally lymphomas). For the comparison with our data, we took into consideration all anomalies of chromosomes 3, 5, 6, 7, 11, 12, and 17 (Table 2; Fig. 1) because attention was drawn to them by various authors or they appeared to be frequently affected in our study. The results on the distribution of the anomalies of these chromosomes are as follows: The two samples of "breast cancer" (literature and our own) are very similar) (x 2 = 0.46, v = 3, nonsignificant), The sum of the two "breast cancer" samples is very similar to the sample "other solid tumors" (×2 = 1.69, v = 4), The sum of all solid tumors is significantly different from the "malignant hemopathies" sample (x 2 = 19.4, v = 6, p ~ 0.005). This difference is principally due to an overinvolvement of chromosome 7 in the "malignant hemopathies" sample (x 2 participation = 5.1, v = 1) and of chromosomes 11 and 17 in the "solid tumors"
+ -
-
5. 6.
7. 8. 9.
+ + +
+ + -
+ +
+ + +
+
+
+ + +
Radiotherapy
34 84 60
168 48
120 45 93
90 75
44
23 73 51
Time primary disease/S-ANLL
+ + -
+
+ + -
+ +
+
+ +
Preleukemia
LAM4 LAM4 ?
LAM2 LAM6
LAM2 LAM2 LAM2
LAM2 LAM2
LAM2
LAM5A ? LAM4
0/5 2/17 ?/O
6/0 0/33
24/46 0/18 0/24
0/29
7/10
0/19
0/10 0/41 0/6
No. normal/ FAB abnormal classification mitoses
(q13.2;qll.1),+r(?) Normal karyotype 53,X,-X,+2,+6,del(17)(p11),+der(1)t(1;8) (q24;q21), + der(1)t(1;8)(q24;q21), +der(8)t(1;8)(q24;21),+ der(5)t(5;14) (p14;q21),+ d e r ( 7 ) , + der(20) 46,XX,- 7,del(5)(p14),+mar 46,XX,t(9;11)(q21;q24) Normal karyotype
44,XX,-5,-7,-16,-17,+der(5)t(5;17)
46,XX,t(lO;11)(p12;q14} 4 6 , X X , - 5 , - 6 , - 17,t(4;15)(q24;q26), t ( 1 I ; 2 1 ) ( p 1 2 . 2 ; p 1 2 ) , + der(5)t(5;17) (q21;q22), + der(6)t(6;?)(q22.1;?),+ m a r 47,XX,+ 8,t(7;12)(q11.2;pter) 47,XX,÷8
+der(22)t(22;?),+ der(22)t(22;?)
(p12 ;ql 1.2), + der(5)t(5;17)(qll ;ql 1),
44,XX,-3,-5,-17,-17,-22,-22,r(11), t(12;13)(p12.1;q13),+der(3)t(3;17)
4 6 , X X , - 7,+der(7)t(7;?}{q22;?} 46,XX,t(8;18)(p21.2;q21.2) 45,XX,-7
Karyotypic formulas
f o r m u l a s f r o m t h e 14 p a t i e n t s w i t h p r i o r b r e a s t c a n c e r ~
° All patients except cases 6 - 8 received a chemotherapy treatment including an alkylating agent (melphalan and/or cyclophosphamide). All radiotherapy treatments were localized to breast area at a total dose of 55 Gy, fractionated in 1.8-Gy courses.
Abbreviations: Ad, adriamycin; Am, ametycin; Bleo, bleomycin; 5-FU, 5-fluorouracil; Eto, etoposide; Met, methotrexate; Ten, teniposide; Vin, vincristine.
12. 13. 14.
10. 11.
Ad, 5FU, V i n Ten, Mef, Bleo Ad, 5FU, T e n , M e t +
Ad, 5FU Ad, 5FU, Eto, M e t Ad, A m , 5 F U Vin, M e t 5FU, M e t
1. 2. 3.
4.
Chemotherapy
Clinical data and karyotypic
Case
Table 1
46
Z. Mamuris et al.
60
50
40
30
20
10
3
5
6
7
ii
12
17
Figure 1 Histograms indicating the participation (in percentage) of chromosomes 3, 5, 6, 7, 11, 12 and/or 17 in the various groups of S-ANLL with prior hematologic malignancy (white), breast cancer, from published data (dark grey) and the present study (black), other solid tumors (light grey), and all solid tumors (hatched).
sample (x 2 participation = 6.5 and 5.2, ~ = 1 for chromosomes 11 and 17, respectively). A d d i t i o n a l information is given by the analysis of the types of rearrangements. Rearrangements affecting the four most frequently i n v o l v e d c h r o m o s o m e s are indicated in Table 3 in relation to the prior malignancy. As previously noted, the complete loss of c h r o m o s o m e 7 is more frequent in S-ANLL with prior hematologic
Table 2
Cases in w h i c h an anomaly of ch r o m o so m es 3, 5, 6, 7, 11, 12 and/or 17 were described
Chromosome
Prior hematologic malignancies
Prior breast cancer
Prior breast cancer (present study)
Prior other solid tumors
Prior solid tumors
3 5 6 7 11 12 17 Sample sizes
9 37 18 60 8 13 14 89
2 8 2 6 3 2 6 19
1 5 2 6 4 1 4 14
6 17 4 17 12 9 15 48
9 30 8 29 19 12 25 81
Source: References [2, 3, 6-8] and present study.
47
Chromosomal Differences in ANLL Table 3
The most frequent chromosome anomalies in S-ANLLa
Anomalies
Prior solid tumors
Prior hematologic malignancies
del(5q) --5 del(7q) -7 del(11) -11 trcp(11;other) del(17p) -17 trcp(17;other) Whole sample size
18 7 8 17 7 0 11 13 6 4 81
17 10 13 41 4 2 0 8 2 2 89
a Unbalanced rearrangements resulting in a deficiencyof the correspondingchromosome are grouped with deletions.
malignancies than with prior solid tumors (x 2 = 16.3, v = 1). Very striking is the high rate of reciprocal translocations of chromosome 11 in S-ANLL with prior solid tumors and its absence in S-ANLL with prior hematologic malignancies. The rates of rearrangements and/or losses of chromosome 5 are very similar in the two samples. DISCUSSION The value of the study of S-ANLL has been stressed by several authors because this leukemia represents a quasiexperimental model for studying a leukemogenic process. Characteristic recurrent anomalies of chromosomes 5 and 7 and, to a lesser degree, of chromosome 17, have already been described [2-11]. The extensive study of Bloomfield [1] concluded that the distribution of anomalies of chromosomes 5 and 7 may vary in relation to the prior malignancy: del(5q) may be characteristic of cases with prior solid tumors, whereas losses of chromosomes 5 and 7, and gains of others chromosomes, would seem to characterize cases with prior hemopathies. Our data on S-ANLL with prior breast cancer are not in contradiction with this proposal: three of three anomalies of chromosome 5q led to deletions (Table 1). However, we also found three cases with loss of chromosome 7, indicating that if the distinction is valid, it is statistical rather than absolute. Additional information on differences between the two samples were found in our study. Chromosomes 5 and 7 were the most frequently affected (in 37% and 34%, respectively, of cases with prior solid tumors). The higher rate of complete loss of chromosome 7 in the "hematologic" versus the "solid tumor" group was confirmed. However, we could not confirm the difference in chromosome 5 involvement as proposed by Bloomfield [1] because this chromosome is equally frequently missing or deleted in both groups of S-ANLL. In the solid tumor group, anomalies of chromosomes 17 and 11 are almost as frequent (in about 29% and 26% of cases, respectively) as those of chromosomes 5 and 7. The frequent involvement of chromosome 17 was previously noticed in S-ANLL with prior solid tumors [7]. Chromosome 11 is principally involved in reciprocal translocations, but no recurrent breakpoints could be found. This excess of lesions of chromosomes 11 and 17 was not observed in S-ANLL with prior malignant hemopathies because they were affected in about 16% and 9% of the cases, respectively, which is not more than several other chromosomes (Fig. 1). It would be of great interest to understand the causes of such differences. A n excess of aberrations affecting chromosome 11 has
48
Z. Mamuris et al. been described after treatment by etoposide and cisplatin [16]. Three of our patients received etoposide or teniposide (cases 2, 9, and 10}, but none of them exhibited any anomaly of chromosome 11 in their leukemic clone. Nevertheless, it remains possible that the type of chromosomal anomalies largely depends on the treatment received for the first malignant disease. Indeed, these treatments differ in the groups with prior solid tumors and malignant hemopathies. The fact that in vitro and in vivo treatments by melphalan {an alkylating agent) induce a high rate of anomalies of chromosomes 11 and 17 [17, 18] and that alkylating agents are quite frequently used in treatment of solid tumors (11 of 14 of our patients; Table 1} may be of importance. However, such drugs are also used in the treatment of hematologic malignancies, and the above explanation may be insufficient for discriminating between the two groups. It would be of great interest to study the respective role of radiation and of the various drugs used for chemotherapy. Unfortunately, the therapeutic protocols vary, and such a comparative study would be premature at the present time. Another element of importance may be the genetic constitution of the patients. In a variety of solid tumors, such as colorectal and breast cancers, rhabdomyosarcomas, and hepato- and nephroblastomas, recurrent deletions and/or allelic losses of chromosomes 11 and 17 have been described [19-25]. This suggests the existence of recessive genes located on these chromosomes, which might be involved in one step of the tumoral process. As indicated by familial studies with recurrent cancers, the abnormal recessive alleles may be transmitted as predisposing factors. Thus, they may exist constitutionally in single copy in a number of individuals. In S-ANLL patients, such genetic determinism can be suspected for recessive genes located on chromosomes 5, 7, 11, and 17, which would be uncovered by deletions of their normal homologs, as we proposed earlier [18]. Thus, it is possible that, among patients with prior solid tumors, several carried abnormal recessive alleles on chromosomes 17p and 11, which might account for the higher involvement of these chromosomes in this group than in that with prior hematologic malignancies. The presence of preexisting mutations may also provide some explanation for the occurrence of SANLL in patients with prior solid tumors treated by surgery alone, as described by Zaccaria et al. [10]. However, this explanation may be insufficient for chromosome 11 because this chromosome is involved in various reciprocal translocations without any visible deletions. The variety of the breakpoints excludes the involvement of a single gene, but it is possible that several genes of this chromosome play a role in tumorigenesis, as suggested by the fact that deletions of the short and long arms are observed in some tumors, such as breast cancer [20]. Another mechanism, recently described, may be also pertinent here: it has been shown that several genes of chromosome 11 are abnormally methylated in a variety of tumor cell lines [26]. If this methylation pattern is determined by a single gene, as seems to be the case for the late replicating X, a relationship may exist between mutagenesis and a multilocus inactivation. This work was supported by grants B16-147F from EURATOM, the Ligue Nationale Fran~aise Contre le Cancer, and the Minist~re de la Recherche et de l'Enseignement Supbrieur.
REFERENCES
1. Bloomfield CD (1986): Chromosome abnormalities in secondary myelodysplastic syndromes. Scand ] Haematol 36 {Suppl 45):82-90. 2. Rowley ]D, Golomb HM, Vardiman ] (19771: Nonrandom chromosomal abnormalities in acute nonlymphocytic leukemia in patients treated for Hodgkin disease and non-Hodgkin lymphomas. Blood 50:759-770.
Chromosomal Differences i n ANLL
49
3. Rowley JD, Golomb HM, Vardiman JW (1981): Non-random chromosome abnormalities in acute leukemia and dysmyelopoietic syndromes in patients with previously treated malignant disease. Blood 58:759-767. 4. Sandberg AA, Abe S, Kowalczyk JR, Zedgenidze A, Takeuchi J, Kakati S (1982}: Chromosomes and causation of human cancer leukemia. L. Cytogenetics of leukemias complicating other diseases. Cancer Genet Cytogenet 7:95-136. 5. Pedersen-Bjergaard J, Philip P, Mortenson RT, Ersboll J, Jansan G, Panduro J, Thomsen M {1981): Acute nonlymphocytic leukemia, preleukemia, and acute myeloproliferative syndrome secondary to treatment of other malignant diseases. Clinical and cytogenetic characteristics and results of in vitro culture of bone marrow and HLA typing. Blood 57:712-723. 6. Pedersen-Bjergaard J, Philip P, Pedersen NT, Hou-Jensen K, Svejgaard A, Jensen G, Nissen NI (1984): II. Bone marrow cytology, cytogenetics, results of HLA typing, response to antileukemic chemotherapy and survival in a total series of 55 patients. Cancer 54:452-462. 7. Groupe Fran~ais de Cytog~n~tique H~matologique (1964): Chromosome analysis of 63 cases of secondary nonlymphoid blood disorders: A cooperative study. Cancer Genet Cytogenet 12:95-104. 8. Arthur DC, Bloomfield CD (1984): Banded chromosome analysis in patients with treatment-associated acute nonlymphocytic leukemia. Cancer Genet Cytogenet 12:189-199. 9. Le Beau MM, Albain KS, Larson RA, Vardiman JW, Davis EM, Blough RR, Golomb HM Rowley JD (1986): Clinical and cytogenetic correlations in 63 patients with therapy-related myelodysplastic syndrome and acute nonlymphocytic leukemia: Further evidence for characteristic abnormalities of chromosomes No. 5 and 7. J Clin Oncol 4:325-345. 10. Zaccaria A, Alimena G, Baccarani M, Billstr6m R, Carbonell F, Castoldi GL, Fuscaldo K, Hecht F, Hossfeld DK, Mitelman F, Rosti G, Sandberg AA, Tascinari A, Testoni N, Tura S (1987): Cytogenetic analyses in 89 patients with secondary hematologic disorders--results of a cooperative study. Cancer Genet Cytogenet 26:65-74. 11. Third MIC Cooperative Study Group (19871: Morphologic, Immunologic and Cytogenetic CMIC) working classification of the primary myelodysplastic syndromes and therapy-related meylodysplasias and leukemias. Cancer Genet Cytogenet 32:1-10. 12. Dutrillaux B, Couturier J (1981): La Pratique de l'Analyse Chromosomique. Masson, Paris. 13. ISCN (1985): An International System for Human Cytogenetic Nomenclature, Harnden DG, Klinger HP (eds.); published in collaboration with Cytogenet Cell Genet (Karger, Basel, 19851; also in Birth Defects: Original Article Series, VoL 21, No. 1 (March of Dimes Birth Defects Foundation, New York, 1985). 14. Hagemeijer A, H~ihlem K, Sizoo W, Abels J (1982): Translocation (9;11}(p21;q23) in three cases of acute monoblastic leukemia. Cancer Genet Cytogenet 5:95-105. 15. Fourth International Workshop on Chromosomes in Leukemia, 1982 (1984): Rearrangements of llq. Cancer Genet Cytogenet 11:294-295. 16. Ratain MJ, Kaminer LS, Bitran JD, Larson RA, Le Beau MM, Skosey C, Purl S, Hoffman PC, Wade J, Vardiman JW, Daly K, Rowley JD, Golomb HM C1987}: Acute nonlymphocytic leukemia following etoposide and cisplatin combination chemotherapy for advanced nonsmall-cell carcinoma of the lung. Blood 70:1412-1417. 17. Mamuris Z, Prieur M, Dutrillaux B, Aurias A (1989): The chemotherapeutic d~ug melphalan induces breakage of chromosomes regions rearranged in secondary leukemia. Cancer Genet Cytogenet 37:65-77. 18. Mamuris Z, Gerbault-Seureau M, Prieur M, Pouillart P, Dutrillaux B, Aurias A (1989): Chromosomal aberrations in lymphocytes of patients treated by melphalan. Int J Cancer 43:80-66. 19. Koufos A, Hansen MF, Copeland NG, Jenkins NA, Lampkin BC, Cavenee WK (!985): Loss of heterozygosity in three embryonal tumors suggests a common pathogenetic mechanism. Nature 316:330-334. 20. Das DD, Schroeder WT, Chao LY, Kikuchi H, Strong LC, Riccardi VM, Pathak S, Nichols W-W, Lewis WH, Saunders GF (1978): Genetic mechanisms of tumor-specific loss of 11p DNA sequences in Wilms' tumor. Am J Hum Genet 41:202-217. 21. Gerbault-Seureau M, Vielh P, Zafrani B, Salmon R, Dutrillaux B (1967): Cytogenetic study of twelve human near-diploid breast cancers with chromosomal changes. Ann Genet 30:138-145. 22. Ali IV, Lidereau R, Theillet C, Callahan R (1987): Reduction to homozygosity of genes on chromosome 11 in human breast neoplasia. Science 238:185-186.
50
Z. M a m u r i s et al.
23. Monpezat JP, Delattre O, Bernard A, Grunwald D, Remvikos Y, Muleris M, Salmon R], Frelat G, Dutrillaux B, Thomas G (1988): Loss of alleles on chromosome 18 and on the short arm of chromosome 17 in polyploid colorectal carcinomas. Int J Cancer 41:404-408. 24. Vogelstein B, Fearon ER, Hamilton SR, Kern SE, Preisinger AC, Leppert M, Nakamura Y, White R, Smits AM, Bos JL (1988): Genetic alterations during colorectal tumor develop,ment. N Engl J Med 319:525-532. 25. Muleris M, Salmon RJ, Zafrani B, Girodet J, Dutrillaux B (1985): Consistent deficiencies of chromosome 18 and of the short arm of chromosome 17 in eleven cases of human large bowel cancer: a possible recessive determinism. Ann Genet 28:206-213. 26. Bustros A de, Nelkin BD, Silverman A, Ehrlich G, Poiesz B, Baylin SB (1988): The short arm of chromosome 11 is a "hot spot" for hypermethylation in human neoplasia. Proc Natl Acad Sci USA 85:5693-5697.