Cytogenetic abnormalities common to adenocarcinoma metastatic to the pleura

Cytogenetic Abnormalities Common to Adenocarcinoma Metastatic to the Pleura Philip T. Cagle, Linda D. Taylor, Mary R. Schwartz, Ibrahim Ramzy, and Frederick F. B. Elder

ABSTRACT: Recently, i(Sq) was suggested to be a nonrandom chromosomal abnormality characteristic of adenocarcinoma of the lung. To further investigate this observation, a chromosomal analysis of five cases of pleural effusions representing metastatic adenocarcinoma from different primary sites (two lung, two breast, and one stomach) was undertaken. The i(Sq) occurred in three of the tumors, one from each of the three different primary sites. In addition, abnormalities of the short arm of chromosome 3 and extra copies of chromosome 7, both of which have been associated with adenocarcinoma of the lung, were simultaneously present in the same three tumors. Our findings demonstrate that i(Sq) is not specific for adenocarcinoma of the lung and that it may have a role in the pathogenesis of adenocarcinomas from multiple organs. The simultaneous presence of i(Sq), abnormalities of 3p, and extra copies of chromosome 7 may indicate a relationship among these abnormalities in multistep carcinogenesis or the development of metastatic potential.

INTRODUCTION A n o n r a n d o m cytogenetic abnormality may be characteristic of a particular cancer and, therefore, is useful as a diagnostic or prognostic marker for that cancer. The acquisition of the chromosomal abnormality is believed to represent a significant step in the pathogenesis of the respective cancer. Examples include the t(8;14) in Burkitt l y m p h o m a , the t(9;22) in chronic myelogenous leukemia, and the del(1 lp) in W i l m ' s tumor. These cytogenetic abnormalities result in the activation of the c-myc and c-abl oncogenes and loss of the c-H-ras oncogene, respectively [1]. Three cases of adenocarcinoma of the lung with i(8q) and three cases with i(9q) have been recently reported, leading to the suggestion that these isochromosomes may be characteristic of adenocarcinoma of the lung [2-4]. In an attempt to determine the validity of this hypothesis, we compared the karyotypes of five malignant pleural effusions representing metastatic adenocarcinomas from different primary sites, lung (two cases), breast (two cases), and stomach Cone case).

From the Departments of Pathology, Baylor Collegeof Medicine and The Methodist Hospital, Houston, Texas, The Institute for Molecular Genetics. BaylorCollegeof Medicine, Houston, Texas, and Vivigen, Inc., Santa Fe, New Mexico, Address reprint requests to: Philip T. Cagle, Department of Pathology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Received December 20, 1988; accepted February 3, 1989.

219 © 1989 Elsevier Science Publishing Co., Inc. 855 Avenue of the Americas, New York, NY 10010

Cancer Genet Cytogenet39:219-225 (1989) 0165-4608/89/$03.50

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Table 1

C o m p o s i t i o n of study sample

Case

Primary site

Age

Sex

Time from original diagnosis

1 2 3 4 5

Lung Lung Breast Breast Stomach

39 52 71 74 37

M F F F M

4 mo 4 mo 1.5 yr 8 yr Same

Adjunct therapy

Postthoracentesis survival

Chemotherapy, radiation Radiation None Radiation None

1 week Alive at 5 mo 3 mo 2 mo Alive at 1 mo

MATERIALS AND METHODS Malignant pleural effusion specimens were obtained by thoracentesis from five patients with previously diagnosed adenocarcinomas of the lung, breast, or stomach (Table 1). After cytologic diagnosis and confirmation of the presence of malignant cells, extra fluid was used for cytogenetic study. A p p r o x i m a t e l y 25 ml of the pleural fluid was spun at 1500 rpm for 10 minutes. The lower 2 ml, i n c l u d i n g the malignant cells, were plated into T-25 cm 2 flasks (1-3 × 106 cells per flask) containing RPMI-1640 m e d i u m and 10% fetal bovine serum. After 24 hours of incubation in 5% CO2, colcemid (0.08 t*g/ml) was a d d e d for 18 hours. Cells were then trypsinized, centrifuged in medium, r e s u s p e n d e d in 0.06 M KC1 for 20 minutes, and fixed in 3 : 1 absolute methanol :glacial acetic acid. The fixed cells were a p p l i e d onto slides, air dried, and aged with dry heat for 20 minutes at 90°C. Banding was performed by a modified trypsin-Giemsa technique T5].

RESULTS I s o c h r o m o s o m e 8q was found in three tumors (cases 2, 4, and 5) representing all three p r i m a r y sites. In addition, abnormalities of the short arm of c h r o m o s o m e 3 and extra copies of c h r o m o s o m e 7 were simultaneously present in the tumors with i(8q).

Lung Adenocarcinoma Case 1. Seven abnormal metaphase cells were analyzed. Five cells were d i p l o i d or

n e a r - d i p l o i d (range 45-49) and two cells were near-tetraploid. The composite karyotype was: 4 5 - 4 9 , X , - X , + d e r ( X ) i ( X p ) , + 1 , - 4 , + d e r ( 4 ) t ( 4 ; ? ) ( q 2 1 ; ? ) , - 1 9 , + m a r . Case 2. A total of seven metaphase cells was analyzed, three normal and four abnormal (Fig. 1). The c h r o m o s o m e n u m b e r ranged from 112 to 202. The quality of the c h r o m o s o m e p r e p a r a t i o n did not permit complete analysis of all chromosomes, Consistent identifiable abnormalities were der(3)t(3;?)(p12;?), i(8q), and three to four extra copies of c h r o m o s o m e 7 per cell.

Breast Adenocarcinoma Case 3. Eight abnormal rnetaphase cells were analyzed. The chromosome n u m b e r ranged from 95 to 126. Less than optimal chromosome m o r p h o l o g y m a d e c o m p l e t e analysis impossible. Consistent identifiable abnormalities were del(1) (p13), i(3q), del(3)(p12), i(5q), der(11)t(11;?)(p;?), der(15)t(15;?)(p;?),

221

Abnormalities Common to Adenocarcinoma

1

li

i

3

f

Figure 1 Partial karyotype of case 2, lung adenocarcinoma, showing the del(3p) and i(8q).

8

Case 4. A total of 11 abnormal metaphase cells was analyzed (Fig. 2). Five cells were

diploid or near-diploid in chromosome number (range 46-48, modal chromosome number 46), three cells were tetraploid, and three contained 84 chromosomes per cell. Consistent structural abnormalities included two copies of a deleted chromosome 1, a derivative chromosome 3 with insertion of unidentified chromosomal material between bands p l l and p13 and deletion of the long arm at band 3q13, unidentified chromosomal material on 5q and 16p, i(8q), and a reciprocal translocation between chromosomes 11 and 14. Consistent numerical abnormalities were loss of one copy of chromosome 2 and gain of one copy of chromosomes 4 and 7.

Figure 2 Karyotype from case 4, breast adenocarcinoma. Arrows indicate abnormalities of chromosomes 3, 7, and 8.

i ll 't I f I I | ii" ni ii l" Ill 6

7

8

13

14

15

9

10

16

11

17

W

19

20

2l

22

12

18

w

XX

222

•. T. Cagle et al.

The composite karyotype was 48,XX,del{1)(p21),+del(1)(p21),-2,+der(3)lpter-~ 3 p 1 3 : : ? : : 3 p l l - - ~ 3 q 1 3 : ) . + 4 , - 5 , + d e r ( 5 ) t ( 5 ; ? ) ( q : ? ) , ~ - 7 , 8,+i(8q),t(ll:14)(q13:f)111, - 16,+ der(16)t(16;?)(p:?).

Stomach Adenocarcinoma Case 5. A total of seven abnormal metaphase cells was analyzed (Fig. 3). The chrom o s o m e n u m b e r varied between 64 and 67. Consistent structural abnormalities were d e l ( l p ) , i(2q) and i(2p), der(3) with an abnormal banding pattern of 3p distal to band p12, der(7) with unidentified material distal to 7q22, (8) with unidentified material at 8q24, i(8q), two der(9) with unidentified material distal to p22, del(1 lp) at band p12: and seven to 14 unidentified marker chromosomes. Consistent numerical abnormalities were extra copies of chromosomes 2, 3, and 7, and loss of one or more copies of c h r o m o s o m e s 8, 14, and 22. The composite karyotype was 64-67,XX, 1,+der(1)t(1;?) ( p l 3 ; ? ) , - 2 , - 2, + i{2 p), + i(2 p), + i(2q), + i(2q), + der(3)t(3 ;?)(p 12 ;?), + der(3 )t(3 :?) (p 12 ;?), + 6, + 7, + 7, + der(7)t(7;?) (q;?),- 8 , - 8, + d er(8),t(8;?), + der(8)t(8:?)(q :?), * i(8q),

F i g u r e 3 Karyotype from case 5, stomach adenocarcinoma. Arrows indicate the abnormalities of chromosomes 3, 7, and 8.

l l

1

21

II

6

i

3

7

8

4

9

10

I¢ 13

14

15

16

8~0 19

2O

21

22

5

11

12

i

|7

17

18

sl XX

,

mar

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A b n o r m a l i t i e s C o m m o n to A d e n o c a r c i n o m a

9 , - 9, + der(9)t(9 ;?)(p22 ;?), + der(9)t(9 ;?)(p22 ;?), + 10, + del(11 )(p 12), - 1 4 , - 14, - 16, + der(16)t(15;16)(?q21;p11),+ 21,del(22}(q13),+ 7mar.

-

DISCUSSION

Isochromosome 8q is suggested to be a nonrandom cytogenetic abnormality characteristic of adenocarcinoma of the lung [4]. Other nonrandom abnormalities believed to be primary in this group of tumors include i[9) [4], extra copies of chromosome 7 [6] and rearrangements of the short arm of chromosome 3 involving 3p14-23 [2-4, 7, 8]. Our findings demonstrate that i(8q) is not specific for adenocarcinoma of the lung and that it is found in adenocarcinomas from multiple organs. Other nonrandom cytogenetic abnormalities have been found to be shared by tumors from multiple organs. These include deletions and rearrangements involving 3p14-23 in lung carcinoma, renal cell carcinoma, ovarian carcinoma, and other tumors [9-12]. Another example is deletions or rearrangements involving 13q14 (the Rb gene) in retinablastoma, osteosarcoma, and small cell carcinoma of the lung [13-15]. In both of these examples, loss of genetic material representing a "tumor-suppressing" gene is a common step in the pathogenesis of more than one type of malignancy [16]. Although we do not know what the specific genetic consequences of i(8q) are, our evidence suggests that the acquisition of i(8q) represents a common step in the pathogenesis of adenocarcinomas in multiple organs. The absence of i(8q) in two

Table 2

Relevant chromosomal abnormalities reported in selected adenocarcinomas Location

Lung t(3;5)(3pter-~cen--~ 5qter}, + 7, + 7 ° t{3;?}(3pter-*cen::?), + 7, + 7 a t(3;14)(3pter-~cen--~ 14qter),del(3)(pter--~q21:), + 7 ° d e l ( 3 ) ( p t e r - * c e n : } , d e l ( 3 ) ( q t e r - * p 2 1 : } , + 7a del(3}(p21),i(8q} b +der(3)t(3;?)(p23 or 24;?),+ 7,i(8q) c + 7,i(8q} d + 7 ( 2 cases) d Stomach +7,i(8q) c del(3)(p21),der(3}t[del(3);?](3pter-*3p23::3p21-*3q23::?} d e r ( 3 ) ( 3 p t e r - ~ 3 p 2 1 : : 3 p 2 6 - - > H S R o n 3p21--~3qter) c i(8q) b +7 b -3,+7 b

Breast + 7,i(8q} c + 7 ( 6 cases) b 3p a b n o r m a l i t i e s ( n u m e r o u s cases} Source of karyotype: Established cell line. Metastasis. c Primary with known metastasis. J Primary, no known metastasis.

Reference

[2] [2] [2] [2] [3] [4] [4] [6]

c

[17] [17] [17] [181 [191 [19] [20] [23] [12, 2 0 - 2 4 ]

224

I'. T. (:~-le et .1

cases indicates that it is not essential ill the pathogenesis of all adenocarcinomas. On the other hand, i(8q) may provide a selective growth advantage to the subpopulation of tunlor cells in which it is present. Because metastatic tumors were studied in this series, the possibility is raised that i(8q) may have a role in the d e v e l o p m e n t of metastatic potential. T wo other c h r o m o s o m a l changes thought to be characteristic of adenocarciimma of the lung, additional copies of c h r o m o s o m e 7 and 3p14-23 abnormalities, were s i m u l t a n e o u s l y present in the three tumors with i(8q). These three ch r o m o so m al abnormalities have also been reported in stomach and breast adenocarcinomas and are often e n c o u n t e r e d in combination and in metastases (Table 2). This suggests the possibility of a relationship among these abnormalities in multistep carcinogenesis or the d e v e l o p m e n t or progression of metastatic potential. The probable role of 3p1423 abnormalities in carcinogenesis has already been mentioned. Possibly, the presence of i(8q) or extra copies of c h r o m o s o m e 7 might result in the activation of an oncogene, w h i c h might contribute to tumor development. The oncogenes c - m y c and c-mos are located on c h r o m o s o m e 8 and A-raf-2, erb B-1 and c - m e t are located on c h r o m o s o m e 7 [25]. The relationship of c h r o m o s o m a l changes to pathogenesis or biologic behavior of carcinomas is often difficult to assess, particularly w h e n numerous abnormalities are present. However, the simultaneous presence of three n o n r an d o m changes in adenocarcinomas metastatic to the pleura from different primary sites suggests that these changes may be c o m m o n to many adenocarcinomas with aggressive potential. The specific role of i(8q) in the d e v e l o p m e n t of adenocarcinoma requires further study. The authors thank Deborah A. Morton and Marsha Kovach for their technical assistance and Janice Bryant for her assistance as transcriptionist.

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