Int. J . Cancer: 45, 50-54 (1990) 0 1990 Wiley-Liss, Inc.
Publication of the International Union Against Cancer Publication de i'Union Internationale Contre la Cancer
CHROMOSOME ABERRATIONS IN METASTATIC OVARIAN CANCER: RELATIONSHIP WITH ABNORMALITIES IN PRIMARY TUMORS M. Josefa BELLO'and Juan A. REY Instituto de Investigaciones Biomidicas del CSIC, Facultad de Medicina de la U.A.M.,Arzobispo Morcillo 4 , 28029 Madrid, Spain. Twenty malignant effusions secondary to ovarian cancer have been cytogenetically analyzed directly and after short in vitro culture. With the exception of one sample characterized by trisomy 3, all cases displayed clonal structural rearrangements. Chromosomes I and 3 were most frequently involved in the genesis of markers. Abnormalities of chromosomes 5, 6, 9, I I and I 2 were also recurrently found, and double minutes (DM) were observed in 2 samples. Our results agree with previous findings on the preferential involvement of chromosomes I, 3 and 6 in ovarian carcinomas, and suggest that rearrangements of certain chromosomes are non-random but are secondary to the malignant progression of these tumors.
A translocation between chromosomes 6 and 14: t(6;14)(q21;q24) was reported by Wake et al. (1980a,b) to be consistently associated with papillary serous adenocarcinoma of the ovary. An involvement of chromosome 6 [mainly del(6q)f has also been reported by Trent and Salmon (1981) and Trent et al. (1985), based on the analysis of human ovarian carcinoma cells cloned in agar. However, other studies (Van der Riet Fox et al., 1979; Woods et al., 1979; Atkin and Baker, 1981; Whang-Peng et al., 1984; Panani and Ferti, 1985; Augustus et al., 1986) have shown that chromosomes 1 and 3 were most frequently rearranged in ovarian tumors. We now present the cytogenetic findings from 20 effusions secondary to ovarian tumors. In addition to aberrations of chromosomes 1 and 3 , recurrent rearrangements of 9p have been observed. MATERIAL AND METHODS
Cytogenetic studies were performed on 20 malignant effusions secondary to ovarian cancer. In 6 cases, 2 samples were analyzed. The clinico-pathological data concerning the 14 patients are summarized in Table I. Chromosome preparations were made directly in all cases and in 8 samples (LPF-97, LPF-151-1 and 2, LPF-173, LPF191, LPF-241-2 LPF-277-1 and 2) additional results were obtained from short-term cultures after 3-7 days of in vitro growth. The methodology used has been described in detail (Rey et al., 1983; Bello et al., 1987); GTL and QFQ banding techniques were used for analysis of karyotypes. RESULTS
A wide variation in the karyotype characterized most of our cases. However, the presence of similar features in all analyzed metaphases from a given sample allowed identification of clonal marker chromosomes characteristic of each tumor. Chromosome findings in the 20 effusions secondary to ovarian tumors are summarized in Table 11. The modal number ranged from 35 to 77, usually in the diploid range. Eight samples showed a hypo-diploid chromosomal mode; in 4 of them it ranged from 35 to 39. One tumor was characterized by 42, 2 by 44 and another by a mode of 45. Both samples from case LPF-277 included a normal diploid sideline; however, in the first sample this side-line coexisted with another hypo diploid (45 chromosomes) which showed the same markers characterizing the stem line. Three samples dis-
played a pseudo-diploid stem line; in all of these, near-diploid secondary cell lines were noted. Hyper-diploid modal numbers were found in 4 tumors, and 3 of them showed clonal structural rearrangements in the stem line. The fourth sample (LPF-173) was characterized by 47 chromosomes, which did not include markers; the sole alteration was trisomy of chromosome 3, and an extra 3 was also found in the cells with 44-46 chromosomes in which random chromosome losses were noted. Five of the 20 samples displayed a modal number in the near-triploid region, with 62, 63, 64, 74 and 77 chromosomes. High ploidy (100 to 270 chromosomes) cell populations were found in all 5 samples. In 6 samples (LPF-128, LPF-131, LPF-173, LPF241-2, and LPF-277-1 and 2) some cells with normal diploid karyotypes were found. With the exception of LPF-173, all cases showed clonal structural rearrangements in the stem line or side lines (Fig. 1). Chromosome 1 was the most frequently rearranged: there were 14 markers (from 9 patients) involving this chromosome. Two of these markers resulted from complex rearrangements in which breakpoints were difficult to identify. Nine markers were derived from deletions implying the loss of a region of lq; the most frequent breakpoint was located at lq32, and the remaining 3 markers were the product of duplication or translocations with variable breakpoints. Figure 2 shows a representative karyotype of case LPF- 191 displaying the characteristic rearrangements, including markers of chromosome 1. Chromosome 3 was involved in the genesis of 10 markers (12 samples from 8 patients). The loss of a region of 3q by deletion or translocation was the abnormality most frequently identified, with breakpoints at q21-23 or q27-28. Ten chromosome 9 derivative markers were present in 6 cases, mainly implying terminal deletions of the short arm at p13 or p22-p23. In only one case was a partial loss of the long arm found: der(9)t(9;?)(q34;?). In Figures 3 and 4, representative karyotypes of both clones of case LPF-277 displaying different 9p deletions are shown. Next in frequency of involvement were chromosomes 5, 6, 11 and 12, which participated in the genesis of 5 markers each. They were rearranged in 4, 4, 4 and 5 cases, respectively. Translocations primarily involved chromosomes 5 , 11 and 12, but no bands of preferential rearrangement were noted. However, loss of a distal region of the short arm of chromosome l l was observed in 3 cases, and involvement of 12~13-12q13 characterized 3 samples. In contrast, deletions, duplications and translocations implying loss of material of the long arm at q15 or q21 involved chromosome 6. Figure 5 shows the distribution of breakpoints for all identified marker chromosomes. In most samples, assessment of the numerical deviations was difficult due to the presence of unidentified marker chromosomes. Gains involving chromosome 3 were seen in 2 samples (LPF-173 and LPF-241-1); this was the only chromosomal
'To whom reprint requests should be addressed. Received: August 28, 1989.
51
CYTOGENETICS OF METASTATIC OVARIAN CANCER
abnormality observed in LPF-173. Also, proportional gains of chromosomes 7, 9, 11, 12, 17, 19, 21 and 22 were present in at least one instance. Regarding chromosome losses, proportional loss of material from chromosome 17 was observed in cases LPF-48 and LPF-277; loss of 11 and 12 characterized 2 samples each, whereas chromosomes from group G(21-22) were involved in proportional losses in 5 cases. Finally, 11 samples displayed loss of the chromosome X. Two of the 20 samples displayed double minutes (DM). Both of them were characterized by a near-triploid modal number. In LPF-151-2, DM were present in 12% of cells. In LPF191, they were present in 60%. The number of DM per cell ranged from 5 to 9 in both samples.
TABLE I - SUMMARY OF THE CLINICO-PATHOLOGICAL DATA FROM THE 14 CASES
Case
niimher
LPF-48 LPF-97' LPF- 106' LPF-108 LPF- 122 LPF- 128 LPF- 13 1 LPF- 132 LPF- 151I LPF- 158 LPF-173 LPF- 191' LPF-241' LPF-277'3'
Age
Histological type
62 58 65
Cystadenocarcinoma Epithelial carcinoma (IV) Endometrioid carcinoma Adenocarcinoma Mucinous-cystadenocarcinoma Papillary adenocarcinoma Epithelial carcinoma Adenocarcinoma Adenocarcinoma Adenocarcinoma Cystadenocarcinoma Endometrioid adenocarcinoma Endometrioid carcinoma I1 Adenocarcinoma
55
66 25 66 39 56 49 33 47 62 56
DISCUSSION
Cytogenetic analysis of direct preparations or after short in vitro growth of primary or metastatic ovarian tumors has shown a variety of chromosomal abnormalities, some of them implying non-random involvement. Therefore, Trent and Salmon (1981) proposed that a relationship exists between ovarian adenocarcinoma and deletions of 6q. Woods et al.
'Two samples were analyzed from these tumors.-2Previous therapy. Survival: 2-12 months.
TABLE I1 - SUMMARY OF CYTOGENETIC FINDINGS FROM ALL THE ANALYZED SAMPLES
Case number
Number of cells analyzed
Representativekaryotype
LPF-48
84
LPF-97-1
57
LPF-97-2 LPF-106-1
39 21
LPF- 106-2 LPF-108
65 49
35,X( -X),del(l)(q42)Ml,de1(3)(p23)M2,-4,-5,-7, - 8,-9, +der(s)t(s;?)(p13;?)M3, - 10,- 11, -15,-16,-17,-17,-19,-20,-21,-22,+5 Mar. 38,XX,- 1,- l,+rea(l)Ml,+der(2)t(2;?)(q34;?)M2,-4,de1(6)(q21)M3,-9,- lO,+der(lO) t(lO;?)(pl3;?)M4, - 11, - 12, - 12, +der(l2)t(l2;?)(pl2;?)M5, - 13, +der(l3)t(l3;?)(q33;?) M6,-14,-15,-18,-20,-21,-22,+2 Mar. '46,XX,- l,+Ml,+M2,M3,-9,- 10, +M4,- 12,+M5,- 13,+M6,- 15,-20,-21,-22,+4 Mar. 38,XX, - 1, +rea(l)(p32q31)Ml, -2,de1(3)(q23)M2, - 5 , -6, - 11, - 11, - 13, - 15, - 17, -21, - 22, 2 Mar. '46,XX,M1,-2,M2,-6,11,+3 Mar. 37,X( -X),del(l)(q32)Ml,del(l)(q24)M2, -2,-2,-3,del(3)(q21)M3, -4,-5,-5,+der(5)t(5;?) (q33;?)M4, -6, -6, +der(6)t(5;6)(q13;ql5)M5,-7, -8,-8,-9,de1(9)(~23)M6, - 10, +der(lO) t( lO;?)(q26;?)M7,- 11, - 11, der(ll)t(l l;?)(p15;?)M8, - 12, der(l2)t(l2;?)(q24;?)M9, - 13, - 14,- 15,- 16, - 18,- 19, +der(l9)t(l9;?)(q13;?)MlO, -20,-21, -22,+ 11 Mar. 46,XX, - 12, +Mar. 47,XX, - 10,+2 M a . 49,XX,+7,-14,+17,+2Mar. 77,XX( - X), - 1, rea(l)(ql2 or q21)Ml ,del(2)(pl3)M2,del(3)(q2l)M3, - 6, - 7, - 7, + der(7) t(7;?)(q34;?)M4, -8,-9,de1(9)(p22)M5, - 10,- 11, +der(ll)t(ll;?)(p14;?)M6, - 12, - 17, - 19, +der(19)t(19;?)(q13;?)M7, -22, 17 Mar. 54,X( -X),del(l)(q32)Ml, +Ml,del(l)(q12)M2, -2,-3, -4,-5,de1(6)(q15)M4,-8,+der(9)t(9;?) (p23;?)M7, - 11, der( 1l)t( 11;?)(pl l;?)MS,del( 1l)(q23)M9, - 12, - 12, + der( 12)t(12;?) (q23;?)MlO,+MlO,- 13,- 14,+der(l5)t(15;?)(~12;?)Mll,- 19,-20,+22,+14 Mar. +M7, +M8,M9, l62,X( -X),Ml, +M1 ,M2,de1(3)(q23)M3, - 5,M4, - 8,del(9)(~13)M5,deI(9)(~23)M6, - 12, - 12, +M10, +M10, - 15, +M11, - 19, +20, -21, 17 Mar. 42,XX, - 1, - 1, +der(l)t(l;?)(p36;?)Ml,de1(3)(p13)M2, -4, -4, - 5 , +der(5)t(5;?)(q34;?)M3, del(6)(421)M4,dup(6)(q22 + 24)M5,de1(7)(q21)M6, + M6, - 8, - 8,de1(9)(p22 or p23)M7, +der(9)t(9;?)(q34;?)M8, +M8, - 10, - 10, - 11, +der(ll)t(ll;?)(q25;?)M9, - 16, - 17, - 18, - 19, -20,-22,+6 Mar. 47,XX, +3. 64,-X, -X, -X, - l,del(l)(q42)Ml, + der(l)t(l;?)(p34;?)M2, del(M2)(q32)M3, - 2, - 2, -2, - 3, - 3, der(3)t(3;?)(q28;?)M4, +der(3)t(?;3;?)(?;~13q27;?)M5,+ del(MS)(p?)MB, -4, -4, - 5 , - 5, -5, +der(S)t(S;?)(plS;?)M7, -6, - 7, -7, -8, -8, - 8, +der(8)t(8;?)(p23;?)M8, - 10, - 10, - 11, - 12,- 12,+der(12)t(12;?)(~13;?)M9,- 14,- 14,- 15,- 15,- 15,t(15q15q)MlO,- 16,+der(l6) t(16;?)(q13;?)Mll, - 17, - 17, - 17, - 18, - 18,-20, -21, -22, +26 Mar.
+
+
+
LPF- 122 LPF- 128 LPF- 131 LPF-132
21 52 41 77
LPF- 151-1
65
LPF-151-2
107
LPF-158
27
LPF- 173 LPF- 191- 1
120 206
LPF- 191-2
24
LPF-24 1- 1 LPF-241-2 LPF-277-1
20 94 67
LPF-277-2
67
+
+
+
+
+
+
'45,-X,-X,Ml,+M2,+M2,-2,-2,-3,+M4,+M6,-4,-5,-5,+M7,-7,-7,-8,-8, +M8,- 10,- 11,- 12,+M9,- 14,- 14,- 15,- 15,+M10,- 1 6 , + M l l , - 17,- 17,- 18,-20, - 21, - 21, - 22, - 22, 18 Mar. 74,XX(-X),+3,-4,+rea(4)Ml,del(lO)(pl2q24)M2,+11,+12,-16,-18,+19,+21,+3 Mar. '63,XX( -X), - 1, -4, + M1, -6, -7, -9, +M2, - 15, - 16, - 18, 2 MU. 44,X( -X), -3, +der(3)t(3;?)(428;?)M3,deI(7)(~13~15)M4, - 8,de1(9)(p22)M8,- 11, - 17, + 2 Mar/ 40,X( -X),-2,-3,de1(9)(p13)M5,11, - 13,- 15,- 15,- 16,- 17, - 19,-21,+5 Mar. '44,X( - X), - 3, + M3 ,M4,M8, - 11, - 17, Mar./40,X( - X), - 2, - 3 ,M5, - 11, - 13, - 15, - 17, -19,-21,+3 Mar.
+
+
+
'The marker chromosomes in these samples are the same as those described in the corresponding first sample. Near-triploid chromosome complements have been defmed as deviations from the perfect triploid karyotype.
52
BELL0 AND R E Y
FIGURE2 - Karyotype of case LPF-191 (marker m12 was not clonal).
FIGURE1 - Clonal marker chromosomespresent in metastatic ovarian carcinomas (see Table I1 for description). Not all marker chromosomes were present in every analyzed metaphase from a given case, and therefore they have been taken from different cells.
(1979) also found 6q- abnormalities in tumor-cell lines derived from patients with serous adenocarcinoma of the ovary, and similar findings have been reported by Kusyk et al. (1982) and Atkin and Baker (1981). Wake et al. (1980) reported that t(6;14)(q21;q24) was consistently associated with ovarian papillary adenocarcinomas, this translocation being present in 6 of the 12 cases studied. In the remaining 6 cases either a 6q - or a 14q+ translocation (or both) were present. However, this has not been confirmed by others (Van der Riet Fox et al., 1979; Woods et al., 1979; Atkin and Baker, 1981; WhangPeng et al., 1984; Panani and Ferti, 1985; Augustus et al., 1986). Whang-Peng et al. (1984) reported abnormalities of chromosome 6 in 17 of the 44 ovarian tumors they studied, but these anomalies implied 6q deletion in 12 instances only. The deletions did not always correspond to papillary serous adenocarcinoma subtype. Markers 14q+ were also seen in 15 samples, nonetheless the extra material was derived from chromosome 6. Five markers derived from chromosome 6 were identified in our series, 3 of them being 6q - , the breakpoint of which was located at bands 6q15 (in 1 instance) and 6q21 (2 instances). These markers were present in different histological types. In addition, no clonal rearrangement involving chromosome 14 was observed. However, in 3 samples, the following deletions were detected: del( 14)(q22) in LPF-97, der( 14)t( 1;14) (p12;q32) in LPF-191 and t(6;14)(q25;q12) in LPF-277; these rearrangements were not clonal because they were found in only one cell each. Our findings do not support Wake’s pro-
posal concerning the specific involvement of chromosomes 6 and 14 [i.e., t(6;14)] (Woods et al., 1979; Wake et al., 1980a,b), but they coincide with the results of Whang-Peng et al. (1984) and Panani and Ferti (1985) which suggest a recurrent involvement of the long arm of chromosome 6 in ovarian tumors. However, keeping in mind that our cases were all metastatic tumors, we could consider the possibility that further complex rearrangements might have occurred secondarily in the markers during the process of tumor progression (Wake et al., 1980b), after the t(6;14) took place, and thus that the derivative der(6), der(l4), or both marker chromosomes may have been difficult to identify in our material. Like others (Van der Riet Fox et al., 1979; Woods et al., 1979; Atkin and Baker, 1981; Whang-Peng et al., 1984; Panani and Ferti, 1985; Augustus et ul., 1986) we have found that chromosomes 1 and 3 were most frequently rearranged. Although chromosome l is most frequently involved in the gen-
FIGURE3 - Karyotype of case LPF-277 showing del(9) and other marker chromosomes.
53
CYTOGENETICS OF METASTATIC OVARIAN CANCER
FIGURE4 - Karyotype of case LPF-277 showing a different 9pmarker chromosome and other structural and numerical deviations. esis of markers in human malignancies (Brito-Babapulle and Atkin, 1981; Atkin, 1986) some reports show recurrent involvement of a certain region of that chromosome in ovarian tumors. Whang-Peng et al. (1984) noted abnormalities of chro-
mosome 1 in 30 of 44 samples, usually as a result of deletions or translocations involving region lq21-q32. Augustus et al. (1986), studying effusions derived from ovarian tumors, also indicated band q21 as the most frequent breakpoint. These findings have been confirmed by Sheer et al. (1987) on permanent cell lines. Eleven of the 14 markers derived from chromosome 1 in our series showed breakpoints in the above regions, generally q32, implying the genesis of markers lq - . Abnormalities of 3p in ovarian tumors have been described. Usually, deletions in 3p13-21 are found (Van der Riet Fox et al., 1979; Whang-Peng et al., 1984; Panani and Ferti, 1985; Hill et al., 1984; Trent and Salmon, 1981; Trent et al., 1985; Sheer et al., 1987). In our series, only 3 of the 10 markers derived from chromosome 3 involved the short arm, one of them at p13. However, deletions of 3q were more frequent in our study, mainly at regions 3q21-23 and 3q27-28. These findings agree with other reports (Bullerdieck et al., 1985a,b; Sheer et al., 1987) which point out that involvement of chromosome 3 in ovarian cancer can occur in a variety of ways, and are similar to the results of Atkin and Baker (1987), who found del(3q) (probably at q21). Table I11 shows a summary of the chromosomes involved in primary and metastatic ovarian tumors from reports published previously. A smaller number of primary tumors has been analyzed. Both materials are characterized by abnormalities of chromosomes 1, 3 and 6, whereas chromosomes 5 , 9, 11 and 12 appear to be mainly rearranged in metastases. Cytogenetic findings on established cell lines also indicate involvement of the same chromosomes (Hill et al., 1984; Wolf et al., 1987; Kunzmann and Holzel, 1987; Sheer et al., 1987). Abnormalities of chromosomes 5, 9, 11 and 12 also appeared recurrently in our study. Alterations of chromosome 5 usually involved the long arm, although we detected no preferentially affected region. Rearrangements of chromosome 12 also involved both p and q arms in our cases. In 2 instances breakpoints were located on the short arm (p12 and pl3), both as a result of translocations. Our results are similar to those of Atkin and Baker (1987), who reported the presence of i(5p) and i(12p) in several primary ovarian tumors. Three of the 5 markers involving chromosome 11 were 1l p , with different
+
TABLE Ill - SUMMARY OF THE CHROMOSOMES INVOLVED IN PRIMARY AND METASTATIC OVARIAN TUMORS. REPORTED BY O T H E R S
Number of cases
Van der Riet-Fox et al. (1979) Wake et al. (1980) Atkin and Baker (1981) Trent and Salmon (1981) Whang-Peng etal. (1984)
FIGURE5 - Location of the breakpoints of clonal rearrangements from the 14 cases.
Bullerdieck et al. (1985a) Panani and Ferti (1985) Augustus et al. (1986) Atkin and Baker (1987) Jenkin and McCartney (1987)
Primary tumors
Nu$kr cases
Metastatic
4
lp,lq,3,13,16, DM
2
1q,3,4,11,17
4
t(6;14), lp, lq
8
t(6;14), lp 1% 3q Dq+
22 3
Dq+ lq, 6q,
22 3
44 7
1% 6q
lq,3p, 3% 6% 9,11p, DM, HSR lq,3,9, DM
54
BELL0 AND REY
breakpoints (pll, p14 and p15). The origin of the additional material could not be determined, but a loss of the distal region of l l p was present in all 3 cases. The location of c-H-ras oncogene at llp15.5 and c-k-ras at 12~12.1could explain the relatively frequent involvement of these chromosomal regions; indeed, amplification and/or mutational activation of this oncogene has been reported in 3 of 17 ovarian tumors (Filmus et al., 1986; Feig et al., 1984; Filmus and Buick, 1985; Yokota et al., 1985). Recurrent alterations of 9p were detected in our cases. A total of 9 different rearrangements were identified, implying the loss of a distal region of 9p: 9pl3-pter and 9p22 or p23pter. Similar rearrangements have also been noted in some ovarian tumors (mainly metastases and permanent cell lines) by Van der Riet Fox et al. (1979), Whang-Peng et al. (1984), Augustus et al. (1986), Kunzmann and Holzel, (1987) and Wolf et al. (1987), showing variable breakpoints but also implying the loss of a distal region. Deletions or displacements of 9p13-21 and 9p24 are also present in malignant gliomas (Rey et al., 1987a,b; Bigner et al., 1988) and several alterations of
9p are also common in some hematological disorders (Bloomfield et al., 1987). Such relatively frequent involvement of 9p in human malignancy may suggest that certain genes important for neoplastic development are present in this region. In summary, our findings suggest that abnormalities of chromosomes 1, 3 and 6 could represent non-random cytogenetic events in ovarian cancers. Losses of 9p, l l p , the involvement of 12p12-ql3 and the presence of DM could be considered as recurrent superimposed secondary changes, perhaps contributing to neoplastic progression, and they are thus more frequent in metastases.
ACKNOWLEDGEMENTS
This work was performed at the Dept. of Genetics, Fundaci6n JimCnez Diaz (Madrid), and supported by grants from the Comisi6n Asesora para la Investigaci6n Cientifica y TCcnica (CAICYT) and from Fundacion Conchita Ribago de JimCnez Diaz.
REFERENCES ATKIN,N.B., Chromosome 1 aberrations in cancer. Cancer Genet. Cytogenet., 21, 279-285 (1986). ATKIN,N.B. and BAKER,M.C., Specific chromosome changes in ovarian cancer. Cancer Genet. Cytogenet., 3, 275-276 (1981). ATKIN.N.B. and BAKER.M.C.. Abnormal chromosomes including small metacentrics in 14 ovarian cancers. Cancer Genet. Cytogenet., 26, 355361 (1987). AUGUSTUS, M,, BRUDERLEIN, S. and GEBHART, E., Cytogenetic and cell cycle studies in metastatic cells from ovarian carcinomas. Anticancer Res., 6, 283-290 (1986). BELLO,M.J., REY,J.A., AVILES,M.J., AREVALO, M. and BENITEZ,J., Cytogenetic findings in an effusion secondary from pleural mesothelioma. Cancer Genet. Cyrogenet., 29, 75-79 (1987). BIGNER,S.H., MARK,J., BURGER,P.C., MAHALEY, M.S., BULLARD, D.E., MUHLBAIER, L.H. and BIGNER,D.D., Specific chromosomal abnormalities in malignant human gliomas. Cancer Res., 88, 405-41 1 (1988). BLOOMFIELD, C.D., TRENT,J.M. and VAN DEN BERGHE,H., Report Of the Committee on Structural Chromosome Changes in Neoplasia. Cytogenet. Cell Genet., 46, 316-324 (1987). BRITO-BABAPULLE, V. and ATKIN,N.B., Break points in chromosome no. 1 abnormalities of 218 human neoplasms. Cancer Genet. Cytogenet., 4, 215-225 (1981). BULLERDIEK, J., BARNITZKE, S., KAHRS,E. and SCHLOOT, W., Further evidence for non-random chromosome changes in carcinoma cells-a report of 28 cases. Cancer Genet., Cytogenet., 16, 33-44 (1985~). BULLERDIEK, J., BARNITZKE, S., KAHRS,E. and SCHLOOT, W., Cytogenetic aberration on two ovarian carcinomas with double minutes, one with a 6q- marker chromosome. Cytobios, 42, 15-24 (1985b). FEIG,L.A., BAST,R.C., KNAPP,R.C. and COOPER,G.M., Somatic activation of ras-k gene in a human ovarian carcinoma. Science, 223, 698701 (1984). FILMUS,J. and BUICK,R.N., Stability of c-k-ras amplification during progression in a patient with adenocarcinoma of the ovary. Cancer Res., 45,4468-4472 (1985). FILMUS,J . , TRENT,J.M., PULLANO, R. and BUICK,R.N., A cell line from a human ovarian carcinoma with amplification of the k-ras gene. Cancer Res., 46, 5170-5182 (1986). HILL, S.M., RODGERS, C.S., HULTEN,M.A. and WILSON,A.P., Cytogenetics of a cell line derived from an ovarian papillary serous cystadenocarcinoma. Cancer Genet. Cytogenet., 12, 321-327 (1984). JENKIN,D.J. and MCCARTNEY, A.J., A chromosome study of three ovarian tumors. Cancer Genet. Cytogenet., 26, 237-337 (1987). KUNZMANN, R. and HOLZEL,F., Karyotype alterations in human ovarian carcinoma cells during long-term cultivation and nude mouse passage. Cancer Genet. Cytogenet., 28, 201-212 (1987).
KUSYK,C.S., TURPENING, E.L., EDWARDS, C.L., WHARTON, J.T. and COPELAND, L.J., Karyotypic analysis of four gynecologic tumors. Gynec. Oncol., 14, 324338 (1982). PANANI,A. and FERTI-PASANTONOPOULOU, A., COmnIOn marker ChrOmosomes in ovarian cancer. Cancer Genet. Cytogenet., 16,65-71 (1985). REY,J.A., BELLO,M.J., DE CAMWS,J.M., BENITEZ,J., AYUSO,M.C. and VALCARCEL, E., Chromosome studies in two human brain tumors. Cancer Genet. Cytogenet., 10, 159-165 (1983). REY,J.A., BELLO,M.J., DE CAMPOS, J.M., KUSAK,M.E. and MORENO, S ., Chromosomal composition of a series of 22 human low grade gliomas. Cancer Genet. Cytogenet., 29, 223-237 (19874. REY,J.A., BELLO,M.J., DE CAMPOS,J.M., KUSAK,M.E., RAMOS,C. and BENITEZ,J., Chromosomal pattern in human malignant astrocytomas. Cancer Genet. Cytogener., 29, 201-221 (1987b). SHEER,D., SHEPPARD,D.M., GORMAN,P.A., WARD,B., WHELAN, R.D.H. and HILL, B.T., Cytogenetic analysis of four human ovarian carcinoma cell lines. Cancer Genet. Cytogenet., 26, 339-349 (1987). TRENT,J.M. and SALMON, S.E., Karyotypic analysis of human ovarian carcinoma cells cloned in short term agar culture. Cancer Genet. Cytogenet., 3, 279-291 (1981). TRENT,J.M., THOMPSON, F.H. and BUICK,R.N., Generation of clonal variants in a human ovarian carcinoma studied by chromosome banding analysis. Cancer Genet. Cytogenet., 14, 153-161 (1985). VANDER RIET-FOX,M.F., RETIEF,A.E. and VANNIEKERK,W .A., Chromosome changes in 17 human neoplasms studied with banding. Cancer, 44, 2108-2119 (1979). WAKE,N., HRESHCHYSHYN, N.M., PIVER, S.M., MATSUI,S.I. and SANDBERG, A.A., Specific chromosome change in ovarian cancer. Cancer Genet. Cytogenet., 2, 87-88 (1980~). WAKE,N., HRESHCHYSHYN, N.M., RVER,S.M., MATSUI,S.I. and SANDBERG, A.A., Specific cytogenetic changes in ovarian cancer involving chromosomes 6 and 14. Cancer Res., 40, 4512-4518 (1980b). WHANG-PENG, J., KUTSEN,T., DOUGLASS, E.C., CHU,E., OZOLS,R.F., HOGAN,W.M. and YOUNG,R.C., Cytogenetic studies in ovarian cancer. Cancer Genet. Cytogenet., 11, 91-106 (1984). WOLF, C.R., HAYWARD, I.P., LAWRIE, S.S., BUCKTON,K., MCCINTYRE,M.A., ADAMS,D.J., LEWIS,A.D., SCOTT,A.R.R. and SMYTH, J.F., Cellular heterogeneity and drug resistance in two ovarian adenocarcinoma cell lines derived from a single patient. In?. J . Cancer, 39, 695702 (1987). WOODS,L.K., MORGAN,R.T., QUINN,L.A., MOORE,G.E., SEMPLE, H.U. and STERMAN, K.E., Comparison of four new cell lines from patients with adenocarcinoma of the ovary. Cancer Res., 39, 44494459 (1979). YOKOTA, J., TSUNETSUG-YOMOTA, Y., BATTIFORA, H., LE FEVRE,C. and CLINE,M.J., Alterations of myc, myb, and ras proto-oncogenes in cancers are frequent and show clinical correlation. Science, 231, 261-264 (1985).
Publication of the International Union Against Cancer Publication de i'Union Internationale Contre la Cancer
CHROMOSOME ABERRATIONS IN METASTATIC OVARIAN CANCER: RELATIONSHIP WITH ABNORMALITIES IN PRIMARY TUMORS M. Josefa BELLO'and Juan A. REY Instituto de Investigaciones Biomidicas del CSIC, Facultad de Medicina de la U.A.M.,Arzobispo Morcillo 4 , 28029 Madrid, Spain. Twenty malignant effusions secondary to ovarian cancer have been cytogenetically analyzed directly and after short in vitro culture. With the exception of one sample characterized by trisomy 3, all cases displayed clonal structural rearrangements. Chromosomes I and 3 were most frequently involved in the genesis of markers. Abnormalities of chromosomes 5, 6, 9, I I and I 2 were also recurrently found, and double minutes (DM) were observed in 2 samples. Our results agree with previous findings on the preferential involvement of chromosomes I, 3 and 6 in ovarian carcinomas, and suggest that rearrangements of certain chromosomes are non-random but are secondary to the malignant progression of these tumors.
A translocation between chromosomes 6 and 14: t(6;14)(q21;q24) was reported by Wake et al. (1980a,b) to be consistently associated with papillary serous adenocarcinoma of the ovary. An involvement of chromosome 6 [mainly del(6q)f has also been reported by Trent and Salmon (1981) and Trent et al. (1985), based on the analysis of human ovarian carcinoma cells cloned in agar. However, other studies (Van der Riet Fox et al., 1979; Woods et al., 1979; Atkin and Baker, 1981; Whang-Peng et al., 1984; Panani and Ferti, 1985; Augustus et al., 1986) have shown that chromosomes 1 and 3 were most frequently rearranged in ovarian tumors. We now present the cytogenetic findings from 20 effusions secondary to ovarian tumors. In addition to aberrations of chromosomes 1 and 3 , recurrent rearrangements of 9p have been observed. MATERIAL AND METHODS
Cytogenetic studies were performed on 20 malignant effusions secondary to ovarian cancer. In 6 cases, 2 samples were analyzed. The clinico-pathological data concerning the 14 patients are summarized in Table I. Chromosome preparations were made directly in all cases and in 8 samples (LPF-97, LPF-151-1 and 2, LPF-173, LPF191, LPF-241-2 LPF-277-1 and 2) additional results were obtained from short-term cultures after 3-7 days of in vitro growth. The methodology used has been described in detail (Rey et al., 1983; Bello et al., 1987); GTL and QFQ banding techniques were used for analysis of karyotypes. RESULTS
A wide variation in the karyotype characterized most of our cases. However, the presence of similar features in all analyzed metaphases from a given sample allowed identification of clonal marker chromosomes characteristic of each tumor. Chromosome findings in the 20 effusions secondary to ovarian tumors are summarized in Table 11. The modal number ranged from 35 to 77, usually in the diploid range. Eight samples showed a hypo-diploid chromosomal mode; in 4 of them it ranged from 35 to 39. One tumor was characterized by 42, 2 by 44 and another by a mode of 45. Both samples from case LPF-277 included a normal diploid sideline; however, in the first sample this side-line coexisted with another hypo diploid (45 chromosomes) which showed the same markers characterizing the stem line. Three samples dis-
played a pseudo-diploid stem line; in all of these, near-diploid secondary cell lines were noted. Hyper-diploid modal numbers were found in 4 tumors, and 3 of them showed clonal structural rearrangements in the stem line. The fourth sample (LPF-173) was characterized by 47 chromosomes, which did not include markers; the sole alteration was trisomy of chromosome 3, and an extra 3 was also found in the cells with 44-46 chromosomes in which random chromosome losses were noted. Five of the 20 samples displayed a modal number in the near-triploid region, with 62, 63, 64, 74 and 77 chromosomes. High ploidy (100 to 270 chromosomes) cell populations were found in all 5 samples. In 6 samples (LPF-128, LPF-131, LPF-173, LPF241-2, and LPF-277-1 and 2) some cells with normal diploid karyotypes were found. With the exception of LPF-173, all cases showed clonal structural rearrangements in the stem line or side lines (Fig. 1). Chromosome 1 was the most frequently rearranged: there were 14 markers (from 9 patients) involving this chromosome. Two of these markers resulted from complex rearrangements in which breakpoints were difficult to identify. Nine markers were derived from deletions implying the loss of a region of lq; the most frequent breakpoint was located at lq32, and the remaining 3 markers were the product of duplication or translocations with variable breakpoints. Figure 2 shows a representative karyotype of case LPF- 191 displaying the characteristic rearrangements, including markers of chromosome 1. Chromosome 3 was involved in the genesis of 10 markers (12 samples from 8 patients). The loss of a region of 3q by deletion or translocation was the abnormality most frequently identified, with breakpoints at q21-23 or q27-28. Ten chromosome 9 derivative markers were present in 6 cases, mainly implying terminal deletions of the short arm at p13 or p22-p23. In only one case was a partial loss of the long arm found: der(9)t(9;?)(q34;?). In Figures 3 and 4, representative karyotypes of both clones of case LPF-277 displaying different 9p deletions are shown. Next in frequency of involvement were chromosomes 5, 6, 11 and 12, which participated in the genesis of 5 markers each. They were rearranged in 4, 4, 4 and 5 cases, respectively. Translocations primarily involved chromosomes 5 , 11 and 12, but no bands of preferential rearrangement were noted. However, loss of a distal region of the short arm of chromosome l l was observed in 3 cases, and involvement of 12~13-12q13 characterized 3 samples. In contrast, deletions, duplications and translocations implying loss of material of the long arm at q15 or q21 involved chromosome 6. Figure 5 shows the distribution of breakpoints for all identified marker chromosomes. In most samples, assessment of the numerical deviations was difficult due to the presence of unidentified marker chromosomes. Gains involving chromosome 3 were seen in 2 samples (LPF-173 and LPF-241-1); this was the only chromosomal
'To whom reprint requests should be addressed. Received: August 28, 1989.
51
CYTOGENETICS OF METASTATIC OVARIAN CANCER
abnormality observed in LPF-173. Also, proportional gains of chromosomes 7, 9, 11, 12, 17, 19, 21 and 22 were present in at least one instance. Regarding chromosome losses, proportional loss of material from chromosome 17 was observed in cases LPF-48 and LPF-277; loss of 11 and 12 characterized 2 samples each, whereas chromosomes from group G(21-22) were involved in proportional losses in 5 cases. Finally, 11 samples displayed loss of the chromosome X. Two of the 20 samples displayed double minutes (DM). Both of them were characterized by a near-triploid modal number. In LPF-151-2, DM were present in 12% of cells. In LPF191, they were present in 60%. The number of DM per cell ranged from 5 to 9 in both samples.
TABLE I - SUMMARY OF THE CLINICO-PATHOLOGICAL DATA FROM THE 14 CASES
Case
niimher
LPF-48 LPF-97' LPF- 106' LPF-108 LPF- 122 LPF- 128 LPF- 13 1 LPF- 132 LPF- 151I LPF- 158 LPF-173 LPF- 191' LPF-241' LPF-277'3'
Age
Histological type
62 58 65
Cystadenocarcinoma Epithelial carcinoma (IV) Endometrioid carcinoma Adenocarcinoma Mucinous-cystadenocarcinoma Papillary adenocarcinoma Epithelial carcinoma Adenocarcinoma Adenocarcinoma Adenocarcinoma Cystadenocarcinoma Endometrioid adenocarcinoma Endometrioid carcinoma I1 Adenocarcinoma
55
66 25 66 39 56 49 33 47 62 56
DISCUSSION
Cytogenetic analysis of direct preparations or after short in vitro growth of primary or metastatic ovarian tumors has shown a variety of chromosomal abnormalities, some of them implying non-random involvement. Therefore, Trent and Salmon (1981) proposed that a relationship exists between ovarian adenocarcinoma and deletions of 6q. Woods et al.
'Two samples were analyzed from these tumors.-2Previous therapy. Survival: 2-12 months.
TABLE I1 - SUMMARY OF CYTOGENETIC FINDINGS FROM ALL THE ANALYZED SAMPLES
Case number
Number of cells analyzed
Representativekaryotype
LPF-48
84
LPF-97-1
57
LPF-97-2 LPF-106-1
39 21
LPF- 106-2 LPF-108
65 49
35,X( -X),del(l)(q42)Ml,de1(3)(p23)M2,-4,-5,-7, - 8,-9, +der(s)t(s;?)(p13;?)M3, - 10,- 11, -15,-16,-17,-17,-19,-20,-21,-22,+5 Mar. 38,XX,- 1,- l,+rea(l)Ml,+der(2)t(2;?)(q34;?)M2,-4,de1(6)(q21)M3,-9,- lO,+der(lO) t(lO;?)(pl3;?)M4, - 11, - 12, - 12, +der(l2)t(l2;?)(pl2;?)M5, - 13, +der(l3)t(l3;?)(q33;?) M6,-14,-15,-18,-20,-21,-22,+2 Mar. '46,XX,- l,+Ml,+M2,M3,-9,- 10, +M4,- 12,+M5,- 13,+M6,- 15,-20,-21,-22,+4 Mar. 38,XX, - 1, +rea(l)(p32q31)Ml, -2,de1(3)(q23)M2, - 5 , -6, - 11, - 11, - 13, - 15, - 17, -21, - 22, 2 Mar. '46,XX,M1,-2,M2,-6,11,+3 Mar. 37,X( -X),del(l)(q32)Ml,del(l)(q24)M2, -2,-2,-3,del(3)(q21)M3, -4,-5,-5,+der(5)t(5;?) (q33;?)M4, -6, -6, +der(6)t(5;6)(q13;ql5)M5,-7, -8,-8,-9,de1(9)(~23)M6, - 10, +der(lO) t( lO;?)(q26;?)M7,- 11, - 11, der(ll)t(l l;?)(p15;?)M8, - 12, der(l2)t(l2;?)(q24;?)M9, - 13, - 14,- 15,- 16, - 18,- 19, +der(l9)t(l9;?)(q13;?)MlO, -20,-21, -22,+ 11 Mar. 46,XX, - 12, +Mar. 47,XX, - 10,+2 M a . 49,XX,+7,-14,+17,+2Mar. 77,XX( - X), - 1, rea(l)(ql2 or q21)Ml ,del(2)(pl3)M2,del(3)(q2l)M3, - 6, - 7, - 7, + der(7) t(7;?)(q34;?)M4, -8,-9,de1(9)(p22)M5, - 10,- 11, +der(ll)t(ll;?)(p14;?)M6, - 12, - 17, - 19, +der(19)t(19;?)(q13;?)M7, -22, 17 Mar. 54,X( -X),del(l)(q32)Ml, +Ml,del(l)(q12)M2, -2,-3, -4,-5,de1(6)(q15)M4,-8,+der(9)t(9;?) (p23;?)M7, - 11, der( 1l)t( 11;?)(pl l;?)MS,del( 1l)(q23)M9, - 12, - 12, + der( 12)t(12;?) (q23;?)MlO,+MlO,- 13,- 14,+der(l5)t(15;?)(~12;?)Mll,- 19,-20,+22,+14 Mar. +M7, +M8,M9, l62,X( -X),Ml, +M1 ,M2,de1(3)(q23)M3, - 5,M4, - 8,del(9)(~13)M5,deI(9)(~23)M6, - 12, - 12, +M10, +M10, - 15, +M11, - 19, +20, -21, 17 Mar. 42,XX, - 1, - 1, +der(l)t(l;?)(p36;?)Ml,de1(3)(p13)M2, -4, -4, - 5 , +der(5)t(5;?)(q34;?)M3, del(6)(421)M4,dup(6)(q22 + 24)M5,de1(7)(q21)M6, + M6, - 8, - 8,de1(9)(p22 or p23)M7, +der(9)t(9;?)(q34;?)M8, +M8, - 10, - 10, - 11, +der(ll)t(ll;?)(q25;?)M9, - 16, - 17, - 18, - 19, -20,-22,+6 Mar. 47,XX, +3. 64,-X, -X, -X, - l,del(l)(q42)Ml, + der(l)t(l;?)(p34;?)M2, del(M2)(q32)M3, - 2, - 2, -2, - 3, - 3, der(3)t(3;?)(q28;?)M4, +der(3)t(?;3;?)(?;~13q27;?)M5,+ del(MS)(p?)MB, -4, -4, - 5 , - 5, -5, +der(S)t(S;?)(plS;?)M7, -6, - 7, -7, -8, -8, - 8, +der(8)t(8;?)(p23;?)M8, - 10, - 10, - 11, - 12,- 12,+der(12)t(12;?)(~13;?)M9,- 14,- 14,- 15,- 15,- 15,t(15q15q)MlO,- 16,+der(l6) t(16;?)(q13;?)Mll, - 17, - 17, - 17, - 18, - 18,-20, -21, -22, +26 Mar.
+
+
+
LPF- 122 LPF- 128 LPF- 131 LPF-132
21 52 41 77
LPF- 151-1
65
LPF-151-2
107
LPF-158
27
LPF- 173 LPF- 191- 1
120 206
LPF- 191-2
24
LPF-24 1- 1 LPF-241-2 LPF-277-1
20 94 67
LPF-277-2
67
+
+
+
+
+
+
'45,-X,-X,Ml,+M2,+M2,-2,-2,-3,+M4,+M6,-4,-5,-5,+M7,-7,-7,-8,-8, +M8,- 10,- 11,- 12,+M9,- 14,- 14,- 15,- 15,+M10,- 1 6 , + M l l , - 17,- 17,- 18,-20, - 21, - 21, - 22, - 22, 18 Mar. 74,XX(-X),+3,-4,+rea(4)Ml,del(lO)(pl2q24)M2,+11,+12,-16,-18,+19,+21,+3 Mar. '63,XX( -X), - 1, -4, + M1, -6, -7, -9, +M2, - 15, - 16, - 18, 2 MU. 44,X( -X), -3, +der(3)t(3;?)(428;?)M3,deI(7)(~13~15)M4, - 8,de1(9)(p22)M8,- 11, - 17, + 2 Mar/ 40,X( -X),-2,-3,de1(9)(p13)M5,11, - 13,- 15,- 15,- 16,- 17, - 19,-21,+5 Mar. '44,X( - X), - 3, + M3 ,M4,M8, - 11, - 17, Mar./40,X( - X), - 2, - 3 ,M5, - 11, - 13, - 15, - 17, -19,-21,+3 Mar.
+
+
+
'The marker chromosomes in these samples are the same as those described in the corresponding first sample. Near-triploid chromosome complements have been defmed as deviations from the perfect triploid karyotype.
52
BELL0 AND R E Y
FIGURE2 - Karyotype of case LPF-191 (marker m12 was not clonal).
FIGURE1 - Clonal marker chromosomespresent in metastatic ovarian carcinomas (see Table I1 for description). Not all marker chromosomes were present in every analyzed metaphase from a given case, and therefore they have been taken from different cells.
(1979) also found 6q- abnormalities in tumor-cell lines derived from patients with serous adenocarcinoma of the ovary, and similar findings have been reported by Kusyk et al. (1982) and Atkin and Baker (1981). Wake et al. (1980) reported that t(6;14)(q21;q24) was consistently associated with ovarian papillary adenocarcinomas, this translocation being present in 6 of the 12 cases studied. In the remaining 6 cases either a 6q - or a 14q+ translocation (or both) were present. However, this has not been confirmed by others (Van der Riet Fox et al., 1979; Woods et al., 1979; Atkin and Baker, 1981; WhangPeng et al., 1984; Panani and Ferti, 1985; Augustus et al., 1986). Whang-Peng et al. (1984) reported abnormalities of chromosome 6 in 17 of the 44 ovarian tumors they studied, but these anomalies implied 6q deletion in 12 instances only. The deletions did not always correspond to papillary serous adenocarcinoma subtype. Markers 14q+ were also seen in 15 samples, nonetheless the extra material was derived from chromosome 6. Five markers derived from chromosome 6 were identified in our series, 3 of them being 6q - , the breakpoint of which was located at bands 6q15 (in 1 instance) and 6q21 (2 instances). These markers were present in different histological types. In addition, no clonal rearrangement involving chromosome 14 was observed. However, in 3 samples, the following deletions were detected: del( 14)(q22) in LPF-97, der( 14)t( 1;14) (p12;q32) in LPF-191 and t(6;14)(q25;q12) in LPF-277; these rearrangements were not clonal because they were found in only one cell each. Our findings do not support Wake’s pro-
posal concerning the specific involvement of chromosomes 6 and 14 [i.e., t(6;14)] (Woods et al., 1979; Wake et al., 1980a,b), but they coincide with the results of Whang-Peng et al. (1984) and Panani and Ferti (1985) which suggest a recurrent involvement of the long arm of chromosome 6 in ovarian tumors. However, keeping in mind that our cases were all metastatic tumors, we could consider the possibility that further complex rearrangements might have occurred secondarily in the markers during the process of tumor progression (Wake et al., 1980b), after the t(6;14) took place, and thus that the derivative der(6), der(l4), or both marker chromosomes may have been difficult to identify in our material. Like others (Van der Riet Fox et al., 1979; Woods et al., 1979; Atkin and Baker, 1981; Whang-Peng et al., 1984; Panani and Ferti, 1985; Augustus et ul., 1986) we have found that chromosomes 1 and 3 were most frequently rearranged. Although chromosome l is most frequently involved in the gen-
FIGURE3 - Karyotype of case LPF-277 showing del(9) and other marker chromosomes.
53
CYTOGENETICS OF METASTATIC OVARIAN CANCER
FIGURE4 - Karyotype of case LPF-277 showing a different 9pmarker chromosome and other structural and numerical deviations. esis of markers in human malignancies (Brito-Babapulle and Atkin, 1981; Atkin, 1986) some reports show recurrent involvement of a certain region of that chromosome in ovarian tumors. Whang-Peng et al. (1984) noted abnormalities of chro-
mosome 1 in 30 of 44 samples, usually as a result of deletions or translocations involving region lq21-q32. Augustus et al. (1986), studying effusions derived from ovarian tumors, also indicated band q21 as the most frequent breakpoint. These findings have been confirmed by Sheer et al. (1987) on permanent cell lines. Eleven of the 14 markers derived from chromosome 1 in our series showed breakpoints in the above regions, generally q32, implying the genesis of markers lq - . Abnormalities of 3p in ovarian tumors have been described. Usually, deletions in 3p13-21 are found (Van der Riet Fox et al., 1979; Whang-Peng et al., 1984; Panani and Ferti, 1985; Hill et al., 1984; Trent and Salmon, 1981; Trent et al., 1985; Sheer et al., 1987). In our series, only 3 of the 10 markers derived from chromosome 3 involved the short arm, one of them at p13. However, deletions of 3q were more frequent in our study, mainly at regions 3q21-23 and 3q27-28. These findings agree with other reports (Bullerdieck et al., 1985a,b; Sheer et al., 1987) which point out that involvement of chromosome 3 in ovarian cancer can occur in a variety of ways, and are similar to the results of Atkin and Baker (1987), who found del(3q) (probably at q21). Table I11 shows a summary of the chromosomes involved in primary and metastatic ovarian tumors from reports published previously. A smaller number of primary tumors has been analyzed. Both materials are characterized by abnormalities of chromosomes 1, 3 and 6, whereas chromosomes 5 , 9, 11 and 12 appear to be mainly rearranged in metastases. Cytogenetic findings on established cell lines also indicate involvement of the same chromosomes (Hill et al., 1984; Wolf et al., 1987; Kunzmann and Holzel, 1987; Sheer et al., 1987). Abnormalities of chromosomes 5, 9, 11 and 12 also appeared recurrently in our study. Alterations of chromosome 5 usually involved the long arm, although we detected no preferentially affected region. Rearrangements of chromosome 12 also involved both p and q arms in our cases. In 2 instances breakpoints were located on the short arm (p12 and pl3), both as a result of translocations. Our results are similar to those of Atkin and Baker (1987), who reported the presence of i(5p) and i(12p) in several primary ovarian tumors. Three of the 5 markers involving chromosome 11 were 1l p , with different
+
TABLE Ill - SUMMARY OF THE CHROMOSOMES INVOLVED IN PRIMARY AND METASTATIC OVARIAN TUMORS. REPORTED BY O T H E R S
Number of cases
Van der Riet-Fox et al. (1979) Wake et al. (1980) Atkin and Baker (1981) Trent and Salmon (1981) Whang-Peng etal. (1984)
FIGURE5 - Location of the breakpoints of clonal rearrangements from the 14 cases.
Bullerdieck et al. (1985a) Panani and Ferti (1985) Augustus et al. (1986) Atkin and Baker (1987) Jenkin and McCartney (1987)
Primary tumors
Nu$kr cases
Metastatic
4
lp,lq,3,13,16, DM
2
1q,3,4,11,17
4
t(6;14), lp, lq
8
t(6;14), lp 1% 3q Dq+
22 3
Dq+ lq, 6q,
22 3
44 7
1% 6q
lq,3p, 3% 6% 9,11p, DM, HSR lq,3,9, DM
54
BELL0 AND REY
breakpoints (pll, p14 and p15). The origin of the additional material could not be determined, but a loss of the distal region of l l p was present in all 3 cases. The location of c-H-ras oncogene at llp15.5 and c-k-ras at 12~12.1could explain the relatively frequent involvement of these chromosomal regions; indeed, amplification and/or mutational activation of this oncogene has been reported in 3 of 17 ovarian tumors (Filmus et al., 1986; Feig et al., 1984; Filmus and Buick, 1985; Yokota et al., 1985). Recurrent alterations of 9p were detected in our cases. A total of 9 different rearrangements were identified, implying the loss of a distal region of 9p: 9pl3-pter and 9p22 or p23pter. Similar rearrangements have also been noted in some ovarian tumors (mainly metastases and permanent cell lines) by Van der Riet Fox et al. (1979), Whang-Peng et al. (1984), Augustus et al. (1986), Kunzmann and Holzel, (1987) and Wolf et al. (1987), showing variable breakpoints but also implying the loss of a distal region. Deletions or displacements of 9p13-21 and 9p24 are also present in malignant gliomas (Rey et al., 1987a,b; Bigner et al., 1988) and several alterations of
9p are also common in some hematological disorders (Bloomfield et al., 1987). Such relatively frequent involvement of 9p in human malignancy may suggest that certain genes important for neoplastic development are present in this region. In summary, our findings suggest that abnormalities of chromosomes 1, 3 and 6 could represent non-random cytogenetic events in ovarian cancers. Losses of 9p, l l p , the involvement of 12p12-ql3 and the presence of DM could be considered as recurrent superimposed secondary changes, perhaps contributing to neoplastic progression, and they are thus more frequent in metastases.
ACKNOWLEDGEMENTS
This work was performed at the Dept. of Genetics, Fundaci6n JimCnez Diaz (Madrid), and supported by grants from the Comisi6n Asesora para la Investigaci6n Cientifica y TCcnica (CAICYT) and from Fundacion Conchita Ribago de JimCnez Diaz.
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