Hrrrdilas 104: 1-13 (1986)

Cytogenetic studies in non-Hodgkin lymphomas-Results from surgical biopsies ULF KRISTOFFERSSONI, SVERRE HEIM', HAKAN OLSSONZ, MANS AKERMAN? and FELIX MITELMAN'

Departments of Clinical Genetics', Oncology2, and Cytodiagnostics?, University Hospital. Lund, Sweden

KRISTOFFERSSON, U., HEIM, S . , OLSSON, H., 4KEiZMAN. M. and MITELMAN. F. 19x6. Cytogenetic studies in non-Hodgkin lymphomas-Results from surgical biopsies. - Heredifas 104: 1-13. Lund, Swe- den. ISSN 0018-0661. Received August 28.1985

Cytogenetic analysis using Giemsa banding technique W ~ S successful in 30 of the 49 adult patients with non- Hodgkin lymphomas investigated. The results are correlated with previous findings in 48 non-Hodgkin lym- phoma patients studied by means of fine needle aspiration in our laboratory. As 8 patients were included in both series, the total number successfully investigatrd was 70. The success rate with both sampling techniques was equivalent, and both methods also seemed to give qualitatively similar information about chromosome pattern. However, cells with a normal diploid karyotype were more frequent in the surgical biopsies.

In the total material, normal karyotypes only were found in 10 patients. In two patients the aberrations were too complex to allow evaluation. The chromosome variation among the remaining 58 cases was dis- tinctly nonrandom. Chromosomes 3 . 7 . 12, and 18 were preferentially gained, whereas chromosomes 1.6, and 14 were most often involved in structural rearrangements. A 14q+ marker chromosome was the single most frequent abnormality; originating through t( 14; 18) (q32;q21) in 10 of 23 cases. The second most com- mon structural aberration was a deletion of the long arm of chromosome 6 with breakpoints at bands q 15 and q21 (12 cases). At least one of the two most common numerical and/or structural changes, +7, + 12, 14q+ and 6q-, were present in 39 of the 58 patients with aberrations (67 YO). Longitudinal studies demonstrated karyotypic evolution during the course of the disease in five of six patients. Simultaneous samples from dif- ferent tumor sites were studied in 10 patients. The findings in 9 cases suggested a monoclonal origin; in one case totally unrelated karyotypes were found in two different lymph nodes at third relapse, suggesting a mul- tifocal origin.

Ulf Kristoffersson, Depurtment of Clinical Generics, Universiry Hospital, S-221 85 Lund, Sweden

We have previously reported our cytogenetic find- ings on fine needle aspirates in 86 patients with non- Hodgkin lymphomas (KRISTOFFERSSON et al. 1985a). In this paper our results obtained by the surgical biopsy technique are presented. The cytogenetic findings in the two series are correlated and discus- sed in relation to data collected from the literature.

Materials and methods A total number of 57 surgically removed samples from 54 adult patients (>15 yrs) admitted to the De- partment of Oncology, University Hospital, Lund, were studied during the period 1979-83. All had a tentative diagnosis of malignant lymphoma; the pa- tients were consecutive in our laboratory.

Fresh tumor biopsy material was obtained from the Department of Clinical Pathology at our hospi- tal. Histopathological examination was made on the

same specimen. A volume of about 0.5 cm' was minced, transferred to a centrifuge tube, and washed once. The cells were then resuspended in McCoy's 5A medium supplemented with 20 YO newborn calf serum, 2 % glutamine, and 2 IU/ml heparin. The cell number was determined and ad- justed to 0.S-2.0~ loh cells/ml. Cultures (usually 10 ml) were incubated over night at 37"C, and treated with colcemid at either a final concentration of 0.0025 pg/rnl over night, or for four hours at a final concentration of 0.1 pg/ml. Preparations were made according to the standard air-drying technique of our laboratory, including hypotonic treatment in 0.075 mol/l KCI and fixation with 4:l methano1:acetic acid followed by three changes in 3: 1 methano1:acetic acid. The chromosomes were stained with a trypsin-Giemsa banding technique.

Whenever possible, 25 metaphases from each sample were analyzed. The chromosomes were clas- sified according to the system recommended by

2 U. KRIST0FI:ERSSON ET AL

ISCN (1978). Only clonal aberrations were consid- ered. A clone was defined as at least two cells with thc same extra chromosome or structural rearrange- mcnt, or three cells with the same missing chromo- some. A stcmline was defined as the most frequent chromosome constitution of the cell population.

All successfully investigated surgical biopsies were numbered in chronological order (101-130). Ten cases also investigated by aspiration sampling, wcre registered according to their aspiration series number (1-86 in KRISTOFFERSSON et al. 1985a).

Histopathological classification was made ac- cording to the Kiel nomenclature (LCNNERT 1978). The abbreviations used are:

CLL Sez Mycfung ic

cblcc cc

cb Ih

ib NOS *

T-cell Mh LI AILD

Hist 1

= chronic lymphocytic leukemia = Sezary’s syndrome = mycosis fungoides = immunocytic lymphoma = centrocytic lymphoma = centroblastic/centrocyticlymphoma

(F = follicular; D = diffuse; F+D = follicular and diffuse)

= centroblastic lymphoma = lymphoblastic lymphoma

= immunoblastic lymphoma = unclassifiable lymphoma according

to the Kiel classification = T-cell lymphoma = malignant histiocytosis = Lennert’s lymphoma = angioirnmunoblastic lymphadeno-

= histiocytic lymphoma

(conv = convoluted cell type)

pathy

Staging was made according to the Ann Arbor clas- sification (CARBONE et al. 1971). R denotes relapse; R1, RII and RIIl = first, second, and third relapse. respectively.

Results Five patients had benign conditions. Among the re- maining 49 patients with non-Hodgkin, non-Burkitt lymphoma, no metaphases or no analyzable meta- phases were observed in 19. Thus, cytogenetic analysis was successful in 30 patients (33 biopsies). ‘Ten of these were also investigated after fine-needle aspiration. As the analysis of the aspirate failed in two patients, eight patients were successfully inves- tigated with both techniques.

Relevant clinical data on the 30 successfully in- vestigated patients are summarized in Table 1. The

Hereditas 104 (1986)

Table 1. Clinical data on 30 patients with non-Hodgkin lymphoma

Case No.

- I22 69 74

116 I I7 126 82 84 85

106 108 129

124 I28 105 38 79 80 83

12s 127 130 101 102

I I8 46

113 I19

I 20

I 07

Sex/ Site Diagnosis Stage age

Sampling at diagnosis (D) i relapse (R)

Mi84 In Fi76 In Fi72 In Mi68 In Fi34 In Fi72 pl Fi67 In Fi69 In Mi69 In Mi80 In Mi65 ts Mi39 In MMI In Fi66 sc Fi71 vt Mi56 In Fi74 In Mi72 In Fi77 th Mi41 111

Fi57 In Mi51 In Mi30 mm Mi38 In Mi34 In Fi7Y In Fi52 sp Mi39 In Fi73 In Mi70 In

CLL I I ic IV ic IVB ic IV ic IV ic IE cbiccF IV cb/ccF IA ch/ccF 111 cb/ccF I1 cb/ccF I I cbiccF I l l cb/ccD I l l chicc IE chicc IE cb IV Ib HIAS Ih IV NOS IV NOS 1A NOS IV NOS I NOS IIE T-cell I l l T-cell I T-cell I1 Mh IV AILD IV AIL13 IV H l S t I I 1

D D D D R D D D D D D D+R D D D D D D K D D D D R D R D R D D

In=lymph nude. pl=pleural exudate. lb=tonsille, sc=suhcutaneouF lymphatic tissuc, vt=vcntricle, th=thyroid, mm =medin~tinal mils>. \ p = s p l e e ~ ~

detailed cytogenetic findings are presented in Table 2. Representative karyotypes from five cases, exemplifying the general quality of the chromo- somal spreads, are presented in Fig. 1-5.

Discussion The discussion in the this paper is based on thc com- bined data from aspiration samples of 48 patients (KRISTOFFEKSSON ct al. 19X5a), and the present re- sults from surgical biopsies. As eight patients were included in both series, a total number of 70 adult patients with non-Hodgkin lymphomas will be dis- cussed. The findings in five children studied during the same period have been reported separately (KRISTOFFERSSON et al. 198Sb).

No other cytogenetic study comparing results from aspiration and surgical biopsies has, to our knowledge, been reported. Analysis on fine needle aspirates of an adequate number of cells (>5X 10’)

Hwcdrtas I04 (1986) NON-HOIIGKIN LYMPHOMAS SURGICAL BIOPSIES 3

7 h h k 2 . Clonal karyotypic changes in 30 patients with non-Hodgkin lymphoma

Case Diagnosis No. of No. of Clonal karyotypicchanges at diagnosis (I)) or relapse (R) No. cells cells

analyzed karyotyped

122 60 73

1 I6 I17 126 82 X 1

8 i I00 1 ox I 2 0

1211 I 2 1 12x 105

3x 79

80 X3

I 2 i 127 I i n I 0 1

I02 107

I I X 1 h

1 1 3 11')

CLL ic

ic

ic cblccF cbiccF

chlccF cbiccF cb/ccF cbiccF

cblccD cblcc cbicc cb

Ib Ib

NOS NOS

NOS NOS NOS T-cell

T-cell 'I -cell

Mh AlLD

AlLD Hist I

1c

IC

23 13 18 II 22 2.5 14 16

11 X 6

28 12 23

7 16 28

33 12

X 10 25

13 4

21 I X

12 6

18 8 7

25 9

8 4 2 8 6

10 7 7

4 4 6 8 9

14 7 8 7

7 4 3

10 in

6 2 5 7

.5 4

6 4 2 8 9

47,XY ,+12,del( 14)(q22)(D) 46,XX,t(l ;?)(p36;?)(D) 46,XX(D) 46,XY ,de1(6)(q21).t(l l;?)(q25;?),t(l5;?).(q26;?)/46,XY(D) 46,XX(R) 49,XX.t( 1 ;14)(p22;q32). +3,+ 12, + 18(D) 46,XX,del(6)(q13),t(14;18)(q32;q21)146,XX(D) 50,XX,+l,de1(3)(q23).+t(7;?)(q36;?),t(7;?)(q36;'?),i(8q),+del(l6)(pl I) ,+r.+mar/

46,XY(D) 46,XY (D) 47,XY ,del(6)(q21),t(l4;?)(q32;?),i(17q)/46,XY(D) 4&49,XY,de1(9)(p13),t(12;?)(p13;?),t( 14;18)(q32;q21).-18.+2-3mar(D) 4X,XY,del(9)(pl3),+9,+lO,t(12;?)(p13;?),t(l4;18)(q32;q2l),+ 17,- 19(R) 47-5~,XY,+2.-4,+12,t(14;lX)(q32;q21 ),t(14;?)(q32;?).+i(17q).+mar(D) 4748,XX,+7.+12.t( 14; 18)(q32;q21)/46,XX(D) 50,XX,+3,+ 16,+18,+mar(D) 43-45,XY,+t( l;?)(p36;?),-6,-8.t( I4;?)lq32;?),deI(l4)(q24),-16.i(17q).+ 18,

464X,XX,+del( l)(q32),+3~(7).t( 14;?)(q32;?),-20,+mar(D) 46,XY (D) 46,XY(D) 46,X,-X,+der(X),t(X;l)(q28;q12),t(3;:')(p27;?),der(12),t(9;12)(ql 1 ;q24(R) 46,XY,i(lq).del(l)(pl~),t(3;?)(q29;?).-5,+del(6)(q2l),-10,t(l2;?)(q24;?),

46,XX(D) .53,XY,+7.+ 17,+5mar(D) 46,XY(D) 46,XY,t(l;1;2)(p22;p36;p14).i(Xq).t(Y;9)(p13;q21),t(l1;19)(q23;q13).

46,XY.del(6)(q15).t(14;?)(q32;?),+mar(D) 46,X,-X,deI(l)(q25),t(Z;l l)(p21;q21).tl:Y; 15)(ql I ;pl I),del(9)(ql I ).-I4.

46,XX(D) 46,XY(RI) 46,XY(RII) 47,XX,+der(2),t(2;3)(pll;qll),t(4;?)(q35;?),+de1(6)(q21),-12(D) 45-51 ,XY,del( I)(p34),-4.-6.+7,+8,t(ll);?)(p15;?).+11 ,t(12;?)(924;?),t(14;?)(q32;?).

46,XX(D)

+ 20, +mar( D)

- 13, i ( 1 Xq)(D)

t( 14;18)(q32;q21)(R)

t( 14;17)(q32;q23).-16,+r,t mar(R)

t(19;?)(p13;?)(D)

was successful in 55 of 80 samples (69 Yo), and with curgical sampling in 33 of 49 biopsies (67 "/o). Thus, the success rates of the two methods were equiva- lent. The findings in 11 patients studied with both techniques arc summarized in Table 3. As can be seen, the general cytogenetic pattern was quite simi- lar. However, normal cells were more common in the surgical biopsies. This is even more apparent in the comparison between the two methods in the total material, summarized in Table 4. Thus, nor- mal diploid clones were present in 16 of 33 (48 %) surgical biopsies, as compared to only 12 of 60 (20 YO) aspiration samples. It may be assumed that malignant cells are preferentially sucked out during aspiration.

To our knowledge, only one longitudinal cyto- genetic study of a lymphoma has been published previously ( KANEKO et al. 1983), demonstrating karyotypic evolution in parallel with changes in morphology. W e have performed longitudinal cytogenetic studies in six patients (Table 5 ) . In none of our cases were any changes in morphology noted during the course of the disease. Clonal karyotypic evolution occurred in all patients, except No. 46, in whom only normal karyotypes were found on three occasions. In the other five, no distinctive evolu- tionary pattern was discernible. The karyotypic changes most commonly seen were gains and losses of whole chromosomes. In no case did any of the characteristic structural aberrations, e.g., 14q+ or

4 U . KRISTOFFERSSON ET AL. Heredifus 104 (1986)

Tuhlu 3. Comparison between the cytogenetic results obtained by aspiration and surgical biopsy

Case Diagnosis Surgical Aspiration No. biopsy sample

71 69 74 82 84 85 38 79 80 83 46

CLL ic ic cb/ccF cbIccF cb1ccF Ib Ib NOS NOS AILD

failure AA NN AN AN NN AA NN AA AA NN

failure AA A A AA failure failure A A AN AA NN NN

A A = abnormal karyotypes only NN = normal karyotypes only AN = normal and abnormal karyotypes

Table 4. Incidences of AA, AN and NN karyotypes in 33 surgical biopsies and 60 aspiration samples

Surgical biopsy

Aspiration sample

NN l l ( 3 3 70) 7(12 %)

AA 17(52 Yo) 48(80 Yo) AN S(1S %) S( 8 Yo)

A A = abnormal karyotypes only NN = normal karyotypes only AN = normal and abnormal karyotypes

Iitble 5. Karyotypic findings in 6 patients studied longitudinally

6q-, appear during the course of the disease, indi- cating that these are in fact primary abnormalities in non-Hodgkin lymphoma. Whether the evolu- tionary changes recorded represent random second- ary events or a predetermined sequential evolution, as has been demonstrated in experimental tumors (MITELMAN 1972), is unknown. The cytostatic treat- ment received by the patients may also be consid- ered in this context. Indeed, karyotypic differences associated with treatment with different cytostatic drugs have been detected in patients with chronic myeloid leukemia (ALIMENA et al. 1979).

Simultaneous investigations of biopsies obtained from two anatomically different tumor sites were performed in eight patients, and in two additional cases bone marrow or peripheral blood was studied in addition to the lymph node samples (Table 6). In general, the findings suggest a monoclonal origin. The minor karyotypic differences detected may all be compatible with random variation of a common stem line. However, a multifocal origin can not be completely excluded. Indeed, this interpretation seems to be more likely in case No. 6. This patient was investigated four times during the course of the disease. Unfortunately, cytogenetic analysis at diagnosis failed. At first relapse the abnormal clone had the karyotype 45-46,XY,-8,+mar. A com- pletely different karyotppe, 46,XY,t(15;?) (pl l ;?) , was found at the second relapse. Samples from two new sites, different from those previously investi- gated, were analysed at third relapse. One sample

<‘a\e Time Sampling Karyotype No. of interval

sampling (mo )

6 RI RII RIII

40 D R

46 D RI RII

62 RI RII

69 D R

129 D R

7 6

8

14 5

7

1

3

4546,XY ,-8, + mar 46,XY,t(15;?)(plI ;?)

4647,XY ,+del( l)(pll),del(S)(q15),de1(6)(p22),-22 4748,XX,+de1(3)(ql I ) , + 17/49,XX,de1(3)(ql I ) , + 16,+ 17 46,XX,t(l;?)(p36;?).-14.-1S,+16,+ 17,-19,+mar/46,XX 46,XY 46,XY 46,XY 49.XX,+3,de1(6)(q15),+7,+lXl49,XX,+3.del(6)(ql~)~~el(9)(q22),+ 12,+ 18 4344,X,-X,-1,-2,de1(2)(p21),+3,~,der(6),-15,der(16),t(l;16)(q12;q24),-17,-18,+21 46,XX,t( 1 ;?)(p36;?),+2,del(17)(p12),t(17;7)(p13;7) 47,XX,t(l;?)(p36;?),+ 12,de1(17)(p12) 48-4Y.XY ,de1(9)(p13),t( 12;?)(p13;?),t( 14;18)(q32;q21).-18,+2-3mar ~,XY,de1(9)(p13).+9,+1O,t(12;?)(p13;?),t(l4;18)(q32;q21),+17.-19

47,XY ,t(15;?)(p11;?),+20

D=Diagnosis. R=Relapse, RI.RlI,RIII = First. second, or third relapse, respectively

Hereditas 104 (1986) NON-HODGKIN LYMPHOMAS: SURGICAL BIOPSIES 5

Fig. 1. Karyotype of Case 122: 47,XY,+12,del(l4)(q22)

Tohle 6. Karyotypic findings in simultaneously studied tumor sites

Case Time Site of Karyotype N o of biopsy

sampling

Rll l

D

D

D

D

D

D

D

D

D

r axilla I neck I groin r groin 1 buccd 1 huccd 1 axilla r axilla I groin I fscl

node PB

I groin

BM

rgroin I groin palate neck I neck I fscl

lymph

47.XY .t( lS;?)(pll;?), +20 4647,XY ,+del(l)(pl l),del(5)(qlS),del(6)(p22),-22 46,XY,t(14;18)(q32;qZl) 46,XY ,t(l2;?)(q24;?),t(l4;18)(q32;q21) 46,XX,der(2),t( 1 ;2)(ql1 ;PI l),de1(3)(p23) 46,XX,der(Z),t(l;2)(qll;pl l),de1(3)(p23) 4647,XY ,t(14;?)(q32;?),+ 16 46,XY 46.XY .de1(6)(qlS) 46,XY,deI(6)(qlS) 51 ,XY,+del(l)(p33q23q25),de1(2)(q14),+de1(3)(p22),

50,XY,+del(l )(p33q23q2S),-2,+del(3)(p22),t(.l;?)(q35;?),+S,+6,+7,+8.+YlS~,XY, +del(l)(p33q23q25).-2,+de1(3)(p22).+4,+S.+7,+8.+ 10-12

*7&83,XY ,-X,-Y ,del( l)(ql l),del( l)(q42),-2,-3,44,+7,-8,-Y ,-lo, + 12,t( 13;?)(q34;?), t(l4;?)(q32;?),t(l4;? )(q32;?),-lS,t(15;?)(q26;? ),-16,-17,-18,-19,-20,-21,-22,+ I-3mar

*8~6,XY,-X,-Y,del(1)(qll),del(l)(q42),-2,-4,-S,~,+7,-8.-8,+Y,-10,-11,+ 12,t(13;?) (q34;?),t(l4;?)(q32;?),t(14;?)(q32;?),t(l5;?)(q26;?),-16,-17,-19,-21.-22,+ IJmar

45 ,X,-Y146,XY 46,XY 5c~52,XY ,t(l;?)(p36;'?),+2,de1(6)(q15),45,de1(9)(p21),+ 12,+ 13,t(14,?)(q32;?),+i(17q),+ 18 47,XY ,t(l;?)(p36;?),t(l4;?)(q32;?),i(17q),+22/46,XY li *80-8S ,XXY Y ,cx/46 ,XY

+4,+S,+7, +8,-12

46,XY

I)-lIiagnosia. RIII=3rd relapse r-right. I=left. fscl=supraclavicular fossa, PB=peripheral blood, BM=bone marrow

karyorype described in relation to the tetraploid level . cx denotes complex unidentifiable karyotypic changes

6 u. KRISTOFFERSSON ETAL.

Fig. 2. Karyotype of Case 84: 50,XX,+ 1 ,de1(3)(q25),+t(3;?)(~26;?),+t(7;?)(q36;?),i(~q),+ 18.

Fig. 3. Karyotype of Case 83: 46,XY , i ( 1 q),+del(6)(q21),-10,t( 12;?)(q24;?) ,-13,+i( 18q).

Hereditas 104 (1986)

Hizrcpditas 104 (1986) NON-HODGKIN LYMPHOMAS: SURGICAL BIOPSIES 7

Fig.4. Karyotype of Case 101: 46,XX,t(1 ;1;2)(p22;p36;pl4),i(8q),t(9;9)(p13;q21),t(11;19) (q3;q13).t( 14; 18)(q32;q21).

8 U KRISTOFFERSSON ET AL Hereditas 104 (1986)

contained the 15p+ marker together with trisomy 20, whereas the other sample presented a com- pletely different karyotype: 46-47,XY,+deI(l) (pll),del(5) (q15),de1(6) (p22),-22. A karyotypic cvolution with loss of the initial changes can, of course, not be excluded, but does seem highly un- likely.

Only six investigations of simultaneously ob- tained samples, not including parallel studies of only bone marrow and peripheral blood, have been previously reported (Table 7). In four cases the re- sults are best explained by a clonal origin. The two remaining cases had completely different karyo- types at different sites. This was particularly striking in the patient reported by FUKUHARA and ROWLEY (1978), in whom a 14q+ marker originated through different mechanisms in different sites, a finding which seems to necessitate an explanation implicat- ing independently arising malignant clones.

Ten patients (14 %) had completely normal karyotypes. The incidence of lymphomas with nor- mal karyotypes reported in the literature varies con- siderably. In the combined data from seven seriesof consecutively studied adult patients only 15 out of 293 cases (5.4 YO) were cytogenetically normal (FLEISCHMAN and PRIGOGINA 1977; MARK et al. 1979; KAKATI et al. 1980; KANEKO et al. 1982a, b, 1983; Y U N I S ~ ~ al. 1982, 1984; BLOOMFIELD et al. 1983), in contrast to 42 out of 261 cases (16 %) reviewed at the FIFTH INTERNATIONAL WORKSHOP ON CHROMOSOMES I N LEUKEMIA-LYMPHOMA 1984 (1986). Our results are thus in good agreement with the workshop data. The important question, of course, is whether the

cells with normal karyotypes represent the malig- nant parenchyma or are merely spontaneously di- viding non-malignant cells. Cytogenetic analysis of lymphomas with normal karyotypes transplanted into nude mice might conclusively answer this ques- tion.

The involvement of chromosomes in numerical and/or structural aberrations is presented in Fig. 6. The chromosomes most commonly affected were 1, 3 ,6 , 12,14, and 18. At least one of chromosomes 3, 6, and 14 were involved in aberrations in 52 of the 60 cases with abnormal clones (87 Yo). The total number of gains and losses of whole chromosomes is presented in Fig. 7. Chromosomes 3, 7, 12, and 18 were most often gained, whereas no obvious pattern was discernible as regards chromosome loss. At least one of the three most common trisomies (3,7, and 12) were present in 22 cases (37 70).

The breakpoints of all structural aberrations have been summarized in Fig. 8. A total number of 177 breaks affecting 86 different bands were found. The distribution is clearly nonrandom. The most com- mon breakpoints (>3 breakdband) were in falling order of frequency (number of cases in brackets): 14q32 (23), 18q21 (lo), lp36 (7), 6q15 (7), 17cen (6), lq11(5), 6q21(5), 3q29 ( 5 ) , 4q35 (4), and 12q24

Deletion breakpointswere especially frequent on chromosomes 1 ,3 ,6 , and 9. The most common de- letions involved the long arm of chromosome 6, with breakpoints at bands 6q15 (7 cases) and 6q21(5 cases). A survey of all 6q deletions in non-Hodgkin lymphomas ascertained from the Catalog of

(4).

7irhle 7. Karyotypes from lymph nodes and other tissues simultaneously investigated in 6 patients with malignant lymphoma reported i n the literature

Reference Diagnosis Tissue Karyotype .

KAKAII et al. HL LN 4X.XY .lp-,+3,6q-,+7,Yp,t( 1 I ;14)(q21 :q32) lY8U BM 46,XY,lp,6q-,t(ll;l4)(q21;q32)146,XY MARKetal. HL LN 50,XY ,t(2;?)(p25;?) .del(2)(pl3),i(3p) ,de1(6)(p21), f7. 1978 +X,+9,+9,-11,-13,+14,-15,+20,i-2I ,-22

LN 44,XY,del(2)(pl3),t(6:?)(p2l:?),del(ll)(ql3),t(l1;14)(ql3;q3l),i(lSq),-20,-22 FLFlSCHMANand CLL LN 46,XY .t(8;14)(q24;432),19p PRlGOGlNA 1977 PB 46,XY ,t(8;14)(q24;q32).19~ FUKUHAKAetal. sc T 48,XY ,-C,-l4,14q+ .21q+?, +4mar I979 BM 4 5 3 - Y FUKUHARAetal. sc LN 48,XX. 1 p+ ,12q+, 14q+ ,+ 18q-(?).+mar I979 BM 47,XX,-7,6q-,7q-,2mar* FUKUWARAand MF LN 48,XX,-9,lp,5q-, +5q-, +Sq-,8q-,9q-. 10p.t(2;8: 14)(q3;q24;q24) KOWLEY 1978 PB 46,XX,t( 1 ;14)(q32;q32)

HL= Histiocyric lympyoma, SC=Srnall cleaved cell, MF=Mycosis fungoides. CLL=Chronic lymphocytic leukemia l.N=Lymph node. BM=Bnne marrow. PB=Penpheral blond, T=Tondla “ I cell only

Hereditus 104 (1986)

14 I

g 0.

f

y 5

7,

!;

NON-HODGKIN LYMPHOMAS SURGICAL BIOPSIES 9

1 * 3 1 5 e 7 8 B 10 I, 12 13 u 15 18 ,I 18 19 20 n 22 x 1

$I

Fig. 6. Total number of chromosomes involved in numeri- cal andlor structural aberrations in 70 cases of non-Hodg- kin lymphoma. The shaded area represents numerical aberrations; the unfilled, structural changes.

Chromosome Aberrations in Cancer (MITELMAN 1985) is presented in Fig. 9. As can be seen, the de- letions have been interpreted as sometimes intersti- tial, sometimes terminal, but a common deleted segment consisting of bands q23-q25 may be dis- cernible. Only one of the 66 deletions ascertained falls outside this region. On the other hand, it may be speculated that all deletions are morphologically identical although this may not be detectable with present cytogenetic techniques. In fact, results re- cently obtained by high resolution analysis of thc 5q- aberration in refractory anemia may indirectly support this interpretation (MITELMAN et al. 1986). Studies with high resolution banding techniques and molecular genetic techniques, in particular in situ hybridization, might help answer this question. Other recurrent deletions affecting chromosomes 1, 3. and 9 clustered to regions 1~31-34, 1q22-24, 1q3243, 3p22-23, 3qll-12, and 9pll-13. None of these occurred as the sole cytogenetic change, and all of them, except del(3)(qll) in Case No. 40, were

found together with other characteristic lymphoma- associated aberrations (see below).

A total of 29 reciprocal translocations were ob- served in 21 patients. Chromosomal breakpoints in- volved more than once were 14q32, l q l l , lp22, and 9q13. All translocations are given in Table 8. The

Table 8. Rcciprocal translocations found in 70 non-Hodgkin lymphomas

Case Sex1 Diag- Translocation No. Age nosis

30 1 0

126 101 62

107 18 37 26 80

107 37

101 41

107 7

24 26 48 55 82

129 120 101 124

in1

I 76 80 -

MI33 Fl78 Fl72 MI38 F170 F179 MI80 MI37 Fl4Y Fl77 MI38 Fl79 MI37 MI38 MISS Fl79 MI5 1 MI61 Fl4Y MI46 Fl68 Fl67 MI39 MI61 MI38 Fl66 MI67 MI78 Fl77

ic NOS ic T-cell ib T-cell ic Ib conv cblccF NOS T-cell T-cell Ib conv T-cell

T-cell cblccF cblccF cb1ccF cblccF cblccF cblccF cblccF cblccD T-cell cblcc cblccD NOS NOS

cc

t(1;4)(qll;q35) t ( I ;2)(qll:p11) t(l;14)(p22;q32) t ( I ; I ;2)(p22;p36;p14) t( I ;16)(ql2;q24) t(2;11)(p21;q21) t(3;4)(q29;pW t(7;9)(pi 1 ; q l l ) t(8;Y)(q13;p24) t(9;12)(q13;q24) t (YmP13;qw t(9;15)(ql I ;plI) t ( l l ; l s ) (p l l ; q l l ) t( 1 I ;19)(q23;q 13) t(13;14)(ql l;q32)

t( 14;18)(q32:q21) t(14;18)(q32;q21)

t(14;18)(q32;q21)

t(14;18)(q32;q21)

t( 14;18)(q32;q21)

t(X;l)(q28;qll)

t ( 14; 17)(q32;q23)

t(14;18)(q32;q21) t( 14;18)(q32;q21)

t( 14;18)(q32;q21) t( 14; 18)(q32;q21)

t( 14;18)(q32;q21)

t(17;18)(plI;qlI) t(lB:Y)(pll ;ql l )

10 U KRlS IOFFERSSON ET AL Hereditas I04 (1986)

Fig. 8. Distribution of breakpoints in 70 cases of non-Hodgkin lymphoma. x inversion, A deletion, 0 isochromosome, reciprocal translocation, nonreciprocal translocation.

Fig. 9. Diagram of deletions in the long arm of chromo- some 6 in non-Hodgkin lymphomas reported in the litera- ture, including the present series. Each vertical line to the right of the chromosome represents the deleted segment of one individual case. Dotted part of line indicates uncer- tainty about the exact breakpoint.

most common breakpoint was 14q32, involved in re- ciprocal translocations in 13 patients, ten of whom had t(14;18) (q32;q21). Nonreciprocal transloca-

tions involving 14q32 were found in 10 additional cases. A similar incidence of 14q32 involvement

WORKSHOP ON CHROMOSOMES IN LEUKEMIA-LYMPHOMA 1984 (1986), whereas in other large series involve- ment of 14q32 has been more frequent, e.g., 71 '% in BLOOMFlELDet al. (1983), and 51 Yo in YuNisct al. (1982,1984). The reason why the series differ in this respect is unknown.

Only one inversion was found, inv(10)(p13q24), present as the sole change in a case of Sezary's syn- drome (Case 68, Fig. 10). Inversions are very rarc in lymphomas; of almost 1300 structural aberrations acertained from the Catalog of Chromosomc Aher- rations in Cancer (MITELMAN 1985), only 18 wcre in- versions. In all instances the inversions were prcscnt together with other clonal aberrations. All 13 inver- sions (11 pericentric, 2 paracentric) with identified breakpoints, including our case, are listed in Table 5).

(44 %) Was found at the FIFTH INTEKNATIONAI

Trrhlr 9 Inversions in non-Hodgkin lymphomas reported in the literature

Inversion Diagnohis Reference

inv( l)(pterp32) Follicular, large cell YUNlSctal. 1984 inv(l)(p13q12) Histiocytic I nv(2)( p I I q2 1 ) inv(3)(q12q27)" Diffuse. large cell, noncleaved YUNlSetal. 1984 inv(h)(p2S?q2 I ?) * inv(4)(p16q21) Large cell, immunohlastic inv(7)(plSql1.2) Diffuse B-cell, mixed inv(9)(p 12q 13?) 1nv(9)( p13q2 I ) Diffuse, mixed-cell YUNlSet al. 1982 inv( IO)(p13q24) inv(l l)(pllq13) Follicular, predominantly small cleaved cell E L O O M F I E L D ~ ~ al. 1983 inv(1 l)(plSq21) Histiocytic MARKetal. 1978 inv(lh)(pl lq24) Histiocytic, nodular K A N E K O ~ ~ ~ ~ . 1982a

-

KAKAI-I et al. 1980 Small cell. lymphocytic B1 OOMFlELDCtal. 1983

Small cell, lymphocytic

Serary's syndrome KRISTOFFERSSON~~ al. 198%

Same case

NON-HODGKIN LYMPHOMAS: SUKGICAL BIOPSIES 11

Fig. 10. Karyotype of Case 6 : 46,XX,inv(lO)(pl3q24).

lsochromosomes were detected in nine cases: i( 17q) in six, i(8q) in two, and i(18q) in one case. All iwchromosomes in malignant lymphomas reported (ref. i n MITELMAN 1985) are provided inTable 10. As in our scries. i(17q) is the most common. This aber-

TO/I /C 10 . Isochromosomes in non-Hodgkin lymphomas reported in the litcrature (ref. in MITELMAN 1985)

lsochrornosome No. of cases

ration is well documented as a frequent, sometimes solitary, change also in a number of other malignant disorders, including acute nonlymphocytic leuke- mia, polycythemia Vera, chronic myeloid leukemia, Burkitt’s lymphoma, acute lymphocytic leukemia, CLL, carcinoma, and malignant melanoma (MITEL- M A N 1985). The two other isochromosomes in our series, i(8q) and i(18q), both occurred concomi- tantly with other changes. A s none of these iso- chromosomes have been observed as the sole change in any neoplasm (MITELMAN 1986), their role seems to be secondary.

All breakpoints in our series are summarized to- gether with similar information from non-Hodgkin, non-Burkitt lymphomas previously reported (MITELMAN 1985) in Fig. 11. The distribution is clearly nonrandom. In this combined material the most common breakpoints (>20 cases) were lp36, 3q29, 6q21, 8q24, llq13,14q32, 17cen, and 18q21. It is striking that 4 of the 8 breakage sites are located in terminal bands. A similarly uneven distribution is also apparent when all the breakpoints are consid- ered: 12 of 30 breakpoints (40 YO) occurring in at least 10 cases arc located in terminal bands, whereas

12 U . KRISTOFFERSSON ET AL. Hereditas 104 (1986)

Fig. 11. Most frequently involved breakpoints in 429 cases of non-Hodgkin lymphoma. + 220breaks

15-19 breaks 3 10-14breaks

1 2 3 4 6 7 9 12 11 10

Fig. 12. Breakpoints in lymphoproliferative disorders. 0 Acute lymphocytic leukemia, A Chronic lymphocytic leukemia, * Burkitt's lymphoma, kitt lymphoma.

Non-Hodgkin, non-Bur-

the expected frequency is 14 % at the 329 band stage (ISCN 1978). This difference is significant (p<0.002, X2-test). Why are terminal bands more often than interstitial involved in structural aberra- tions? It could be speculated that the telomeric reg- ions contain genes of importance for the malignant process andlor are more prone to undergo chromo- somal rearrangements. However, it can not be excluded that these findings merely reflect the shortcomings of present cytogenetic techniques in identifying interstitial structural abnormalities.

MITELMAN (1984, 1986) demonstrated that only a limited number of breakpoints were regularly invol- ved in chromosomal aberrations in human neopla- sia, and postulated that these regions might contain genes of special importance for the carcinogenic process. Furthermore, the data indicated that

breakpoints of malignant diseases affecting similar cell types might cluster to specific chromosomal re- gions. Since heterogeneity in this respect may be present also among morphologically related types of malignant diseases, the breakpoints have been recorded in all lymphoproliferative disorders repor- ted with a single structural abnormality as the only change from the normal diploid karyotype ( M I T ~ L - MAN 1985). The breakpoints of all identical structu- ral aberrations present in at least two cases, subdivi- ded according to diagnostic group, are provided in Fig. 12. The material is undoubtedly too small to permit any definite conclusions, but the distribution of breakpoints is compatible with a heterogeneity among different lymphatic disorders. This interpre- tation may be, supported by the good agreement be- tween the breakpoints in the malignant lymphomas

Hrrrditas 104 (1986) NON-HODGKIN LYMPHOMAS. SURGICAL BIOPSIES 13

o f Fig. 12 and those of our cases. Thus, at least one of the 11 breakpoints was involved in 33 of our 59 ca- ses with structural aberrations; in addition, 2 cases showed engagement of an adjacent band. Among the remaining 24 cases, 12 had at least one, usually several, unidentified marker chromosomes that may have involved one of the relevant breakpoints. Excluding these, 33 of 47 patients (70 YO) had an en- gagement of at least one of these particular chromo- somal regions. In view of the difficulties in exactly localizing breakpoints to specific bands in malignant lymphomas, the results indirectly provide a strong argument favoring a nonrandom pattern of chromo- some variation in non-Hodgkin, non-Burkitt lym- phomas.

Acknowledgements. -This work was supported by grants from the Swedish Cancer Society, the JAP Foundation for Medical Re- warch. and from the Medical Faculty, Lund University.

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