GENES, CHROMOSOMES & CANCER 3:338-345 (1991)

Cytogenetics of Synovial Sarcoma: Presentation of Ten N e w Cases and Review of the Literature J. Limon, K. Mrozek, N. Mandahl, B. Nedoszytko, A. Verhest, J. Rys, A. Niezabitowski, M. Babinska, H. Nosek, T. Ochalek, A. Kopacz, H. Willen, A. Rydholm, S. Heim, and F. Mitelman

Departments of Biology and Genetics u.L,K.M., B.N.,M.B.) and Oncologicai Surgery (A.K.), Medical Academy, Gdansk and Departments of Pathology (JR, A.N., T.O.)and Surgery (H.N.), Center of Oncology, Krakow, Poland; Departments of Clinical Genetics (N.M., S.H.,F.M.), Pathology (H.W.), and Orthopedics (A.R.), University Hospital, Lund, Sweden; Department of Medical Genetics, Odense University, Denmark (S.H.); and Department of Pathology, Erasme Hospital, Free University, Brussels, Belgium (A.V.)

Cytogenetic study of five biphasic and five monophasic synovial sarcomas revealed the specific abnormality t(X; 18) (p I I;q I I) in eight cases and t ( X 15; 18) (p I I ;q I5;q I I) and t(X;7) (q I I I2;q32) in one case each. Additional, secondary aberrations were present in eight of these tumors. By combining our data with information on previously published cytogenetically abnormal synovial sarcomas, we were able to evaluate 32 tumor samples from 29 patients. The modal chromosome number was pseudodiploid or near diploid in 26 of the 32 tumors. A t(X,18) was present in 21 of 29 cases (72%). Complex translocations involving chromosomes X and I 8 and another autosome were present in five cases, and one displayed a t(5; 18). There was no visible rearrangement of chromosome bands Xp I I or 18q I I in only 2 of the 32 synovial sarcomas. Half of the primary tumors (6 of 12) had the X 18-translocation as the sole abnormality. O f the remaining 20 specimens from recurrent or metastatic tdmors (in three cases t w o tumors could be analyzed), only one had t ( X 18) as the sole change. The secondary aberrations in cases exhibiting clonal evolution were also generally more extensive in the metastatic and recurrent than in the primary sarcomas (five additional aberrations per case, compared with two).Chromosomes I and I 2 were the chromosomes most frequently (one fourth of the cases) involved in additional structural changes, but with several different breakpoints. No differences were identified between the karyotypic profiles of monophasic and biphasic synovial sarcomas.

-

INTRODUCTION

Synovial sarcomas account for approximately 10% of all soft-tissue sarcomas. They are classified histologically as monophasic and biphasic tumors (Krall et al., 1981; Abenoza et al., 1986; Souls, 1986 Enzinger and Weiss, 1988). Biphasic synovial sarcomas are composed of epithelial elements arranged in glandlike structures scattered against a uniform spindle-cell background. The proportions of spindle cells and epithelial elements vary from tumor to tumor. Biphasic tumors and monophasic fibrous synovial sarcomas, the latter being spindle-cell tumors without an epithelial component, are diagnosed with almost equal frequency; monophasic epithelial tumors are rare. Synovial sarcomas are characterized by the specific chromosomal abnormality t(X18) (pl1;qll) (Limon et al., 1986 Griffin and Emanuel, 1987; Smith et al., 1987; Turc-Care1et al., 1987;Wang-Wuu et al., 1987).Sometimes, variants of the standard t(X18) have been found, and additional aberrations indicative of clonal evolution are also detected. However, the limited number of cases that were analyzed cytogenetically has thus far precluded any meaningful attempts to correlate the cytogenetic findings with clinico-pathologic data. Our aim in the present study was, therefore, not only to present ten new karyotypically abnormal synOVid sarcomas, but also to look for COrrekitiOnS be0 1991 WILEY-LISS, INC.

tween the cytogenetic, clinical, and pathologic findings in these and previously published cases. M A T E R I A L S AND M E T H O D S

Excised tumor tissue from ten patients was obtained directly after surgery or after overnight transportation in Hanks’ balanced salt solution supplemented with antibiotics. Table 1 presents the clinico-pathologicdata. Except for case 2, none of the patients had received radio- or chemotherapy prior to sampling. The tumors were primaries in five cases, local recurrences in four, and a metastasis in case 5. A previous metastasis from this case has also been characterized cytogenetically (Verhest et al., 1991), and a previous local recurrence from case 9 has been analyzed (Limon et al., 1989). Cases 3 and 6-10 were analyzed in Gdansk, cases 1and 4 in Lund, and cases 2 and 5 in Brussels. The tumor specimens were processed for cytogenetic analysis as described by Limon et al. (1989) (cases 2, 3, and 5 1 0 ) or by Mandahl et al. (1988a) (cases 1 and 4). Chromosomes were G-banded with Wright’s stain. The aberrations were classified according to the ISCN (1985). Received November 13, 1990; accepted March 20, 1991. Address reprint requests to Dr. Janusz Limon, Department of Biology and Generics, 1 Debinki str., 80-211 Gdansk, Poland.

339

CYTOGENETICS OF SYNOVIAL SARCOMA

TABLE I. Clinico-Pathologic Data on Ten Cases of Synovial Sarcoma

Age/ Sex

Histologic SubtYPea

Location

Date of Diagnosis (month/ Year)

I 2 3 4 5

7YF 20/M 3S/M I3/M 3 3/F

Mo Mo Mo Mo Bi

Forearm Forearm Thigh Mediastinum Knee

7/88 8/89 I0/88 11/86 3/83

6 7 8

23/M I6/F 53/M

Bi Bi Bi

Foot Knee Foot

6/89 1/89 9/83

9

26/M

Bi

Thigh

1/85

10

32/M

Bi

Foot

2/88

Case No.

Date of Recurrence (R) or Metastasis (M)

M: 11/88 R 9/87 R: 8/88 M: 8/89 4/90 7/90 9/90

R 5/88 3/89 R 4/86 1/87 11/87 4/88 11/89 R 2/90 M: 5/90

aMo, monophasic; Bi. biphasic.

Fig. I. Karyotype from case 3 showing t ( X I8)(pl I .2;q I I.2). The r(I ) and monosomies of chromosomes 5 and I I are additional, secondary aberrations. Arrowheads indicate breakpoints.

Outcome Alive 9/90 Alive 6/90 Alive 6/89 Dead 12/87 Dead 10190

Alive 7/90 Alive 1/90 Alive 5/90 Alive 12/89

Alive 5/90

340

LlMON ET AL.

TABLE 2. Cytogenetic Findings in Ten Cases of Synovial Sarcoma

Case No. I 2 3

Number

Histologic Subtypea

Source of Sampleb P P

Mo Mo Mo

P

of Metaphases 7 19 22 II

4

R

Mo

6

3

5

MoC

M

40

6 7 8

Bi Bi Bi

P P R

20 20 16

9

Bi

R

4

3 2

10

R

Bi

II

Karyotype 46,X,t(X18) (pl I;ql I) 47,Y.+E,t(X;I8) (pl I;ql I) 45,Y. - I , -5,- I I, + 12,t(X18) (PII .2;q I I .2), +r(l) (:p12+cen+q44:)/ 44,Y, - I, - 5, - I I,t(X 18) (p I I .2;q I I .2), r( I) (:q44 +cen +p I2::q44+cen +p I2:) 45,Y.t(X;l8) (pl I;ql l),t(l;?)(p36;?), dic(5;2 I) (p I5;p I3),t(7; 13) (p I5;q I2)/ 46,Y,+21,t(X18) (pl I;ql I), t(l;?) (p36;?),dic(5;21) (p I5;pl3).t(7; 13) (p I5;q I2),dic( I5;2l) (p I3;p I I) 46,X,t(X;7) (q I Io r I2;q32),t(7; 14) (q22;q I I) t(8; 12) (q22;q24) 46,Y,t(X;15;18) (pl I;ql5;ql I) 48,X,+7,+ 15,t(X;18) (pl I;ql I) 47,Y,+2,t(X;18) (pl I;ql l),t(4;22) (q22;p I l),t(7; 19) (93 I;p I I), t(12;13) (q13;pl I) 45,Y, - 12, - I7,t(X; 18) (p I I ;q I I). + der( 12) t(12;17) (PI I;ql I)/ 48,Y,+8,+ 10,- 12,- 17,+21,t(X18) (PII;ql I), +der(12)t(12;17) (pl I;ql I)/ 88,polyploid with one copy of der( 12) t(12;17) (pl I;ql I) and one copy of der(l I)t(l 1;17) (pl I;ql I) 36.Y,- l,-3,-4,10,- I I,- 12,- 13,- 15,- 16, -20,-21,-22,t(X;I8) (pl I;ql I),inv(I I) (p 12q l4), + der( I2)t( 12; 13) (p I3;q 12). der(22)t( I;22) (q I2;p I I)

+

+

aMo, monophasic; Bi, biphasic. b ~ primary; . R, recurrence; M, metastasis. 'Primary tumor was biphasic.

For immunohistologic studies, the peroxidase-antiperoxidase (PAP) technique was used on paraffin sections or fresh material. Daco K 518 and Daco M 613 kits were used for detection of keratin and epithelial membrane antigen (EMA),respectively. Electron microscopy was performed in cases 3, 7, and 8. RESULTS

Clonal chromosome aberrations were detected in all ten tumors (Table 2). A t(X18) (pl1;qll)was found in eight tumors; in case 3, it was possible to localize the breakpoints at subband level to Xp11.2 and 18q11.2 (Fig. 1).In case 6, there was a variant translocation, t(X;15;18) (pll;ql5;qll)(Fig. 2), and in case 5, a t(X;7) (qll-l2;q32), with no visible rearrangements of Xpll or 18ql1,was present. Only a single karyotypic anomaly was found in two primary tumors, one monophasic and one biphasic (cases 1 and 6). The primary

tumors of cases 2 and 7, in addition, had simple numerical changes, and the fifth primary tumor (case 3) had both numerical aberrations and an r(1) (p12q44) (Fig. 1)as evidence of clonal evolution.The five recurrent or metastatic tumors had more secondary abnormalities (Fig. 3). Chromosome 12 was involved in secondary translocations in four of the tumors, but always with different breakpoints. Chromosomeband 13q12 was secondarily rearranged in two tumors (cases 4 and 10).

Fig. 2. Complex variant translocation, t ( X IS; 18)(pl I;q I5;qI I), from case 6. Arrowheads indicate breakpoints.

CYTOGENETICS OF SYNOVIAL SARCOMA

34 I

Fig. 3. The five recurrent o r metastatic synovial sarcomas all had additional karyotypic abnormalities indicating clonal evolution. In this karyotype (case 4), the secondary aberrations t( l;?)(p36;?), dic(5;2 I) (p I5;p 13). and t(7; I3)(p l5;q 12) are seen in addition to the primary t(X; I8)(p I I;q I I); - I 4 is a nonclonal change. Arrowheads indicate breakpoints.

DISCUSSION

The finding of a t(X18) (p1l;qll)in eight of the ten tumors in the present series emphasizes the nonrandom occurrence of this translocation in synovial sarcomas. The detection of the variant t(X;15;18)in one of the two remaining cases further strengthens this conclusion. As a result of the addition of these ten cases, 32 specimens from primary and recurrent or metastatic synovial sarcomas from 29 patients were available for evaluation (Table 3). In this larger group, t(X18) was present in 21 patients (72%), complex translocations involving chromosomes X and 18 and another autosome-namely, 1,12,15(twice),and 21were present in five, and t(5;18)was found in one case. There was no visible involvement of X p l l or 18qll in only two synovial sarcomas with karyotypic abnormalities. The karyotypic profiles did not differ between monophasic and biphasic synovial sarcomas. The entire cancer cytogenetics literature of 14,141 neoplasms with clonal abnormalities (Mitelman, 1991) contains only two tumors with a t(x;l8) (pl1;qll) and a diagnosis that was not synovial sarcoma: one was a fibrosarcoma (Mandahl et al., 1988b) and the other, a

malignant fibrous histiocytoma (Turc-Carel et al., 1987). Given the difficulties involved in diagnosing soft-tissue sarcoma, one cannot rule out the possibility that these tumors, also, might actually have been synovial sarcomas, albeit with somewhat different histologic features. At any rate, the consistency with which X;18 rearrangements are found in both monophasic and biphasic synovial sarcomas is quite remarkable; both the specificity and the sensitivity of t(Xl8) make it a useful diagnostic marker for this tumor type. Six of 12 primary tumors among the total of 32 specimens included here had an X;18 translocation as the sole cytogenetic abnormality (Table 3). Among the six primary tumors with evidence of clonal evolution, a simple additional numerical aberration was found in three cases, one tumor had an additional structural aberration, and two tumors had both structural and numerical secondary changes. The recurrent or metastatic tumors typically had more secondary changes than did the primary tumors; in only one recurrence was t(X;18)the sole anomaly (Table 3). Also, the additional aberrations were more complex in the recurrent tumors and metastases than in the primaries; on the

Mo

Bi Bi Bi Bi

I3/M

?/F

28/F

24/M

34/F 30/M

I5/M

25/F

3 I/M 23/M I6/F I7/F

I3/F

4

13

14

15

16 17

18

19

20 6 7 21

22

II 2 3

Bi

Mo

Buttock

Tongue Foot Knee Knee

Lung

Lung

Lung Lung

Lung

Mo

Mo Mo

Thigh

Pelvis

Mediastinum

Forearm Calf Foot Forearm Thigh

Site

Mo

Mo

Mo

Mo Mo Mo Mo Mo

72/F I YF 24/F 20/M 35/M

I 12

Subtypeb

Anel -. Sex

Case No.”

P

P P P P

M

M

M M

M

R

R

R

P P P P P

TissueC

+

+

+

+

46 46 48 a.47 b.45 46

+7,+ 15 - x, + 7, + 9 -X

+ + + +

+4.+7,+9,+ 12, 12,+ 13,+ 15, 20, 2 I , 2 I

+4

b.47 56

+4

+ I5

+21

I I,+ 12

II

Numerical

+8 -5,-5,-

a.47

46 47

47

46

+ I

46

b.46

46 46 46 47 a.44 b.45 a.45

Modal Chromosome Number‘

+

Prior TheraPYd

t ( X 18) t ( X 15; 18) t(X; 18) t ( X 18) t(X; 18) t(X; 18)

t ( X 18)

t ( X I; 18)

t ( X I; 18)

t ( X 15; 18) t(X; 18)

t(X 18)

t(X; 18)

t(X; 18)

t(X; 18)

t(X, 18) t(X, 18) t(X; I8;2 I) t(X; 18) t(X; 18) t(X; 18) t(X; 18)

Structural

ins( 15; I I)

I;?)* der(2)t(2; 12). t( I I ;?),del(12) t( I;I ;6),t(6; lo), inv( I2),der( 18) t( I;I ;6),t(6; lo), der( 18) t(7;9)

t( I;3),t(

del(3)

t(7; 13) t( I;?),dic(5;21) t(7;I 3),dic( I5;2 I) t(4;?),t(5; I2), del(5) t(X;3),de1(6), del(7),t( I2;?), t( 12; I7),t(22;?) inv(4),de1(8), der( I7)t(8; I7),

t( I;?),dic(5;21)

r(l)

dic( I;I)

Chromosomal Aberration

TABLE 3. Clinico-Pathologic and Cytogenetic Data on 29 Cases (32 Tumors) of Synovial Sarcoma

Ueda et al. (I988)

Bridge et al. (I 988) Present series Present series Gregoire et al. (I989)‘

Turc-Carel et al. (I 987)

Wang-Wuu et al. (I 987)‘

Turc-Carel et al. ( 1987) Turc-Carel et al. (I 987)

Limon et al. (I 986) Turc-Carel e t al. (I 987)

Smith et al. (I 987)‘

Smith et al. (I 987)‘

Present series

Present series Griffin and Emanuel (I 987) Turc-Carel et al. (I987) Present series Present series

Reference

PD PD nd

Calf Arm Mediastinum

P R M

M 47 49 44

a.45 b.46 46

44

90

36

cases 5, 9, and 23, two different tumors were analyzed. bMo, monophasic; Bi, biphasic; PD, poorly differentiated; nd. not determined. ‘P, primary: R. recurrent; M, metastatic. dPrior therapy denotes chemo- and/or radiotherapy. ‘a