Chromosome Translocation in Three Cases of Human Malignant Melanoma1

(CANCER RESEARCH 46, 1526-1529, March 1986]

A t(1;19) Chromosome Translocation in Three Cases of Human Malignant Melanoma1

Annette H. Parmiter,2 Gloria Balaban, Meenhard Herlyn, Wallace H. Clark, Jr., and Peter C. Nowell

Departments of Dermatology and Pathology [A. H. P./VV. H. C.], Human Genetics [G. B.], Pathology and Laboratory Medicine [P. C. N.], University of Pennsylvania School of Medicine, and the Wistar Institute [M. H.], Philadelphia, Pennsylvania 19104

ABSTRACT advantage apparently conferred by this chromosomal rearrange ment. Abnormalities of chromosome 1, including trisomy for all or a portion of the long arm, have been frequently reported in many cancers. Anomalies of chromosome 19 are far less common, MATERIALS AND METHODS although a t(1;19Xq23;p13) translocation has been reported in association with pre-B-cell leukemia. We have observed a We have obtained chromosomal data from cell lines or direct prepa rations of 28 cases of advanced human malignant melanoma (9).3 The t(1;19Xq12;p13) translocation in three cases of advanced mela cell lines were treated with colchicine for 30 min at 37Â°C.After treatment noma, with the translocation chromosome representing an extra with a KCI-sodium citrate hypotonie solution and standard fixation, air dose of 1q in each instance. The breakpoint on 1q was within dried slide preparations were made for Giemsa banding (10). For direct the centromeric heterochromatin, proximal to the site in pre-B- preparations the tumor was minced to obtain a single-cell suspension, cell leukemia, but the breakpoint on 19p appeared identical. The treated with colchicine for 60 min at 37Â°C,and processed as described gene for human insulin receptor has recently been mapped to above. Karyotypes were determined from a minimum of 15 counts and this region of chromosome 19 (p13.2-13.3). This gene shares 3 banded karyotypes for each case. structural and sequence homologies with the epidermal growth C-banding studies were carried out, using a modified protocol of factor receptor (erb-B oncogene) and members of the src family Arrighi and Hsu (11), on two of the three tumors with the t(1;19) of oncogenes, suggesting that alterations in the insulin receptor, translocation (WM 39, ML793, and ML991 ) (Table 1). resulting from chromosomal translocation, could lead to a role in tumorigenesis. The present findings may permit this possibility RESULTS to be examined in a neoplasm of neuroectodermal origin. The chromosome findings in the three cases reported are INTRODUCTION summarized in Table 1. Cells from the primary lesion of case WM 39 were first examined at passage 64 in culture. The modal Specific chromosomal translocations have been shown to be number was hypotetraploid, and all karyotypes had a associated with certain types of human cancers (1, 2). It has t(1 ;19)(q12;p13) translocation which resulted in an extra dose of been hypothesized that these nonrandom karyotypic changes 1q (the translocation was not reciprocal). The cells also had extra involve sites in the genome where genes important in carcino- copies of chromosome 7, a 7p+ marker of undefined origin, a genesis are located ("proto-oncogenes") (3). This hypothesis has t(10;13)(q11 ;q34) translocation, and a 16q+ marker (Fig. 1). This found support from studies of several hematopoietic tumors cell line was examined on two other occasions over the course (e.g., Burkitt's lymphoma and chronic granulocytic leukemia) of 7 months and was karyotypically stable. where specific chromosome translocations have been shown to Cells from case ML793, derived from an advanced primary result in altered function of the c-myc and c-abl proto-oncogenes, lesion, were first examined at passage 5 in culture. The modal respectively (4, 5). number was hyperdiploid, and the t(1 ;19Xq12;p13) translocation Nonrandom involvement of certain chromosomes, particularly (Fig. 1, inset) again did not appear to be reciprocal, resulting in Nos. 1,6, and 7, has been observed in human malignant mela an extra dose of the long arm of chromosome 1. There were noma (6-9). The region of chromosome 1 most commonly in variable changes in chromosome 6 and an extra copy of chro volved is 1p11-p22, suggesting that a gene important in the mosome 7, plus other random markers. These cells were again development of this neoplasm is located within the proximal examined at passages 11 and 41 with retention of the t(1;19) portion of 1p. We report three cases of advanced melanoma translocation and the appearance of several new alterations with a t(1;19Xq12;p13.3) translocation. Since the breakpoint is (Table 1). not within the region 1p11 -p22 previously reported in melanoma, In the third case, ML991, four metaphases were obtained from a different mechanism is probably responsible for the selective a direct preparation of a metastasis to the skin. The one cell karyotyped had 69 chromosomes including four t(1 ;19Xq12;p13) Received 9/15/85; revised 11/20/85; accepted 11/21 /85. translocation chromosomes, (Fig. 1, inset), an extra copy of The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in chromosome 7, a del(6)(q21), and additional undefined markers. accordance with 18 U.S.C. Section 1734 solely to indicate this fact. The same markers could be identified in the three other meta ' This research was supported by Grants CA-25298 and CA-25874 from the phases from this preparation. A second skin metastasis, National Cancer Institute. 2To whom requests for reprints should be addressed, at University of Pennsyl (ML991A), removed at the same time, was established in tissue vania, Department of Dermatology, 247 Medical Education Building/GM, Philadel phia, PA 19104. 3Unpublishedresults.

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Table 1 Chromosomal data from the 3 cases with the t(1;19) translocation no.Nn of chromosome chro nfkaryotypes chromo CaseWM39ML793ML991LesionPrimaryPrimaryMetastasis mosomeno.8585854849496979797979Abnormalities15 somechangest(10;13),16q+,*6t(10;13),16q+,*t(10;13),16q+,***2p-,iso7p,*1t8p+,*8p+,*8p+,*8p+,* (64)"9/8411/846/83 +1,+t(1;193 6q-)) +1,+t(1;192 +1,+t(1;193 6q-)II)

(5)7/83(11)3/84(41)9/84 +1,+t(1;195 +1,+t(1;194 +1,+t(1;191

1Metastasis (direct)11/8411/84(5)12/845/85Counts34162118154345241714Modal+1,+t(1;193 del6q21) 2Date4/84 +1,+t(1;191 del6q21) +1,+t(1;193 del6q21) +1,+t(1;193 del6q21) +1,+t(1;196) del 6q217+7,+7p++7,+7p++7,+7p++7+7+7+7+7+7+7+7Additional a Numbers in parentheses, passage in culture. 6 *, undefined markers. K

\\\fr UM mi u nu 10 11 12

*M â€¢â€¢â€¢Mil 15 WM39n\ ML 793 ML991 16 17 18 fftM I 19 20 fiÂ»r 21 22 Fig. 1. Karyotype from melanoma cell line WM 39 demonstrating two der t(1 ;19Xq12;p13) chromosomes (arrow), as well as characteristic 7p+, 16q+, t(10;13X11 ;q34) translocation and four unidentified markers. This metaphase had only 75 chromosomes due to random loss. Inset, der t(1;19Xq12;p13) chromosomes from another WM 39 cell and from the two other melanomas, ML793 and ML991, in which this rearrangement was identified. culture and subsequently karyotyped after 2,3, and 7 months in findings support the conclusion from the G-banded material that culture. These cells showed the same consistent alterations as the breakpoint was within the centromeric region of chromosome the karyotype from the direct preparation, with the addition of 1. an iso8q appearing at 7 months (Table 1). In order to clarify the location of the breakpoint on the long DISCUSSION arm of chromosome 1, C-banding was done on cases ML793 and ML991. In both cases, as illustrated in Fig. 2, two C-banded Frequent involvement of chromosomes 1, 6, and 7 has been areas were present on the der(1;19) chromosome, apparently reported in human malignant melanoma (6-9). This report of corresponding to the active centromere of the No. 19 and a three cases of a t(1;19) translocation, however, apparently rep portion of the centromeric heterochromatin of the No. 1. These resents the first example of a nonrandom translocation in this

CANCER RESEARCH VOL. 46 MARCH 1986 1527

of a similar finding in other leukemias and nonhematopoietic cancers (23). Taken together, these observations suggest that 19 p13 is the site of a gene important in carcinogenesis. Recently, the gene for the human insulin receptor has been mapped to this region (24). Since this gene shares structural and sequence homologies with the epidermal growth factor receptor (the enb-B oncogene) and with the products of the src family of oncogenes, it has been postulated that it may also act as an oncogene (24). Rearrangement of this gene may alter its normal receptor function and thereby contribute to neoplastic prolifera tion. A fragile site has also recently been mapped to chromosome 19 and reported as 19 q13 (25). Given the difficulty of proper arm assignment in chromosome 19, this locus could actually be at 19 p13, the site involved in the breakpoints described above. The significance of the common fragile sites in the genome is still unknown, but it has been speculated that they may play a role in oncogenesis (26, 27). Melanoma cells carrying the particular t(1;19) translocation observed here may prove useful for exploring these various questions concerning 19p and 1q at the molecular level.

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Annette H. Parmiter, Gloria Balaban, Meenhard Herlyn, et al.

Cancer Res 1986;46:1526-1529.

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