Cytogenetic study of twenty-three primary squamous cell carcinomas of the lung

ELSEVIER

Cytogenetic Study of Twenty-Three Primary Squamous Cell Carcinomas of the Lung Song-Bin Fu and Pu Li

ABSTRACT: Fifty-seven primary squamous cell carcinomas of the lung were analyzed cytogenetically. Karyotyping was possible in seven cases, and chromosome counting without detailed analysis was possible in 16 other cases. The results suggested that structural chromosome rearrangements related to the short arms of chromosomes 1(5/7), 9(3/7), and 11(6/7), and the long arms of chromosomes 6(4/7) and 7(6/7) may be the primary and non-random chromosome defects which are closely associated with human lung squamous cell carcinoma. These primary and non-random chromosome defects are believed to confer a proliferative advantage to cells carrying them, and to be involved in the pathogenesis of human lung squamous cell carcinoma. © Elsevier Science Inc., 1997

INTRODUCTION

Lung cancer is one of the more c o m m o n h u m a n cancers and the third leading cause of cancer death in either sex in China. Bronchogenic carcinomas consist of four major histologic s u b t y p e s - - s q u a m o u s cell carcinomas, adenocarcinomas, undifferentiated large cell, and small cell lung carcinomas (SCLC). The squamous cell carcinomas, a d e n o c a r c i n o m a s , and undifferentiated large cell lung carcinomas are also called n o n - s m a l l cell lung carcinoma (NSCLC). In the field of cytogenetic study of h u m a n lung cancer, a n u m b e r of t u m o r cell lines h a d been a n a l y z e d [1, 2] a n d some c h r o m o s o m e defects of the p r i m a r y lung cancers were found [3-8]. But the specific c h r o m o s o m e changes of the p r i m a r y lung cancers were not observed because of the small case n u m b e r s and poor karyotypes analyzed. This report presents the cytogenetic analysis of seven p r i m a r y squamous cell carcinomas of the lung. MATERIALS A N D METHODS

P r i m a r y tumors of 57 s q u a m o u s cell lung carcinomas were obtained from patients of the Division of Thoracic Surgery, the First a n d S e c o n d Affiliated Hospitals of Harbin M e d i c a l University. The diagnoses of the tumors were established on clinical and histologic data. The patients received no preoperative c h e m o t h e r a p y or radiotherapy. The p r i m a r y lung cancers were a n a l y z e d cytogenetically

From the Laboratory of Medical Genetics, Department of Biology, Harbin Medical University, Harbin 150086, P.R. China

Address reprint requests to: Dr. Song-Bin Fu, Laboratory of Medical Genetics, Harbin Medical University, Harbin 150086, Peoples Republic of China. Received June 22, 1996; accepted December 7, 1996. Cancer Genet Cytogenet 9 9 : 5 4 - 5 8 (1997) © Elsevier Science Inc., 1997 655 Avenue of the A m e r i c a s , N e w York, NY 10010

using a m e t h o d established in our laboratory. Fresh t u m o r samples were m i n c e d with scissors and disaggregated, first for 20 m i n in 0.5% t r y p s i n at 37°C, then for another 10-20 m i n u t e s in a fresh 0.5% trypsin solution. The cells were d i s p e r s e d in culture m e d i u m and s p u n down, then r e s u s p e n d e d in fresh m e d i u m and a l l o w e d to s e d i m e n t for 20 minutes. Thereafter, both the s e d i m e n t and supernatant were p l a t e d in culture flasks. The growth m e d i u m was RPMI 1640 w i t h HEPES buffer s u p p l e m e n t e d with 15% fetal calf serum, glutamine, and antibiotics. After culture at 37°C for 12-48 hours, the cultures were arrested by 1-hour exposure to Colcemid. The cultures were harvested using standard 0.075 M KC1 h y p o t o n i c shock, m e t h a n o l / a c e t i c acid fixation, and air drying. G-banding was done with Wright stain. RESULTS

Fifty-seven cases of p r i m a r y squamous cell lung carcinomas were s t u d i e d in short-term culture preparations before patients received any treatment. Karyotyping was possible in seven cases, and c h r o m o s o m e counting without detailed analysis was possible in 16 other cases. In the r e m a i n i n g 34 cases, no mitotic cells were observed. Table 1 shows the clinicopathologic data of the seven cases w i t h detailed karyotyping. Chromosome counting s h o w e d three cases in the h y p o d i p l o i d range (44-45), three cases in the h y p e r t e t r a p l o i d range (94-112), eight cases in the hyperd i p l o i d range (47-58), and nine cases in the h y p e r t r i p l o i d range (73-90). Of the seven cases w i t h G-banding, chrom o s o m e gains and losses were i n v o l v e d in almost all chromosomes. Chromosome rearrangements related to c h r o m o s o m e s 1, 6, 7, 9, and 11 were found in almost all of the seven cases w i t h G-banding. Of these structural abnormalities of c h r o m o s o m e 7 were noted in six of the seven

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Study of Primary S q u a m o u s Cell Carcinomas

Table 1 Case

55

Table 2

Clinical and histologic data of patients Sex/age (yr)

Tumor size (cm)

Node involvement

Metastases

1

M/53

3x 2 X2

+

-

3

M/60

3 X4 × 3

+

-

6°

M/58

2 X3X3

+

+

6b

Case 1

~pl.5

19

F/42

1 X 0.5 × 0.5

-

-

21

M/36

~p0.4

-

-

49

M/5O

1 X 1 × 0.5

-

+

53

M/57

4 X6 × 5

+

+

aKaryotype from the primary tumor of case 6. bKaryotype from the metastases of case 6.

6°

cases, l p - in five, 6q- in four, 9p- in three, and 11p- in six. Detailed cytogenetic findings are s u m m a r i z e d in Table 2. The karyotypes from seven cases are s h o w n in Figures 1-4.

6a

DISCUSSION Most solid tumors have structural a n d / o r n u m e r i c a l chrom o s o m e aberrations, some of w h i c h are consistently associated w i t h particular types of tumors. Up to now, detailed cytogenetic data of the h u m a n s q u a m o u s cell lung carcinoma, especially primary s q u a m o u s cell lung c ar ci n o m a [9], have been sparse. Zech et al. analyzed cytogenetically two cases of lung s q u a m o u s cell c a r c i n o m a and found c h r o m o s o m e rearrangements i n v o l v i n g l p and a translocation b e t w e e n c h r o m o s o m e s 22 and Y [10]. In a cytogenetic study of two poorly differentiated h u m a n lung squamous cell c a r c i n o m a lines, Law et a]. [11] n o t e d three shared marker c h r o m o s o m e s in the cell lines, del(1)(q11), del(2) (p11.1) and del(2)(q11.1). We used our i m p r o v e d chromosomal preparation t e c h n i q u e for solid tumors to analyze some types of h u m a n cancers and found several non-rand o m c h r o m o s o m e rearrangements [12]. In the present study, three cases (cases 19, 21, and 49) w i t h early stages of lung cancer s h o w e d h y p o d i p l o i d karyotypes (Table 2) and deletions of 6q, 7q, and 11p, w h i c h might represent prim ary c h r o m o s o m e changes in lung squamous cell carcinoma. Rearrangements related to 6q, 7p, and 11p also existed in four other cases (cases 1, 3, 6, and 53) w i t h late stages of lung squamous cell carcinoma, possibly indicative that these marker c h r o m o s o m e s m a y be p r i mar y and p e r m a n e n t c h r o m o s o m e changes. In case 6, the karyotypes (Table 2 and Fig. 2) were successfully obtained from the prim ary t u m o r and its metastases. The n u m e r i c a l chromosome aberrations, especially c h r o m o s o m e gains i n v o l v i n g c h r o m o s o m e s 2, 3, 7, 8, 10, 11, 14, 16, 17, 18, and 21, constituted i m p o r t an t differences b e t w e e n the karyotypes of the p r i m ar y t u m o r and that of its metastases in case 6. This suggested that n u m e r i c a l changes of some chromosomes might play a role in the metastasis of h u m a n lung cancer. The h o m o g e n e o u s stain region (HSR) f o u n d in

Clonal n u m e r i c a l and structural alterations in seven primary lung squamous cell carcinomas

19 21 49

53

Karyotype 62-76,XX,-Y,- 1,del(1)(q12)× 2,del(1)(p12) x 2,del(1) (p31),add(1)(p22),der(1)t(1;22)(p11;q12),+ 2,ins(3) (p22p21)× 2 , - 4 , - 5 ,- 5,+6,der(6)t(6;11)(p24;p13), + 7,add(7)(p 12),- 8 ,- 8,add(8)(p?) x 2 ,i(8)(q10) x 2, + 9,- 10,- 11,add(11)(p?)- 12,add(14)(p?)- 15, - 1 5 , - 1 6 , - 1 7 , - 1 8 , - 1 8 , - 18,+19,-20,-21, + 22, + 2mar[cp8] 47-54,X,-Y,del(1)(q22),del(1)(p22),der(1;7)(p10;q10), + 2,del(3)(q12),-4,-4,i(5p),i(6p),del(6)(q21) x 2, - 8,+ 9, +9,add(10)(pll),- 11,del(11)(p11),del(12) (q22),- 13,- 13,- 1 4 , - 1 5 , - 15,-16,+ 19,- 21,-21, -22,-22,+4mar[cp12] 54-81,-X,-Y, + 1, + 1,del(1)(p22),- 2,i(3p),del(3) (p11)×3,-4,- 5 ,- 5,- 5 ,- 6,del(6)(q23),del(7)(p15), add(7)(q23),der(8;9)(q10;q10),+ 9,der(9;11)(q10;q10), - 10,- 10,add(11)(q23),del(11)(p11),- 12,- 13,- 13, - 1 3 , - 1 4 , - 15,- 15,- 1 6 , - 1 7 , - 18,-18,+ 19,-20, - 2 1 , - 2 2 , - 2 2 , + 12mar[cp5] 85-96,XY,-Y,+ 1,inv(1)(p13p35),del(1)(p22),- 2, der(2;3)(p10;q10),del(3)(q11) × 2,del(4)(q22),- 5, i(5p),- 6,del(6)(q16),+ 7,+ 7,add(7)(q12),- 8,- 8, - 9 , - 1 0 , - 10,add(10)(q?),+ 11,- 12,- 1 2 , - 1 3 , - 13, - 13,- 1 4 , - 1 5 , - 15,-16,add(16)(q?),- 17,- 18,-19, - 2 0 , - 2 0 , - 21,-21,-21,-21,+18mar[cp6] 44-49,XX,- 6,del(7)(qllq22),- 9 ,- 10,der(11)del(11) (p12)del(11)(q23),- 18,- 19,- 20,+4mar[cp19] 42-51 ,XY,del(6)(q23),- 7, - 13, - 21, + 2mar[cp18] 45-47,X,-Y,der(1;11)(q10;p10),ins(3)(p14p21),- 5, del(6)(q12),del(6)(q21),i(8q),- 11,- 12,- 14,- 15, - 17,-18,-21,-22,+3mar[cp10] 47-56,X,-Y,del(1)(q31),del(1)(p12),add(1)(p?),+ 3, - 4 , - 4 , + 6,del(7)(qllq22),- 8,hsr(9)(q11),i(9q), del(11)(p11),- 12, + 14,- 15,add(15)(p?),- 16, - 16,+ 17,+20,-22,+3mar[cp15]

°Karyotype from the primary tumor of case 6. bKaryotype from the metastases of case 6.

case 53 i n d i cat ed that amplification of some genes might be associated w i t h the progression of the carcinoma. In conclusion, structural c h r o m o s o m e rearrangements related to the short arms of c h r o m o s o m e s 1 (cases 5, 7), 9 (cases 3, 7), and 11 (cases 6, 7), and the long arms of chrom o s o m e s 6 (cases 4, 7) and 7 (cases 6, 7) m ay be the primary and n o n - r a n d o m c h r o m o s o m e defects that are closely associated with h u m a n lung squamous cell carcinoma. These primary and n o n - r a n d o m c h r o m o s o m e defects are b e l i e v e d to confer a proliferative advantage to cells carrying them, and to be i n v o l v e d in the pathogenesis of the hum an lung squamous cell carcinoma.

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S.-B. F u a n d P. Li

1

: :i:i i il . . . . .

2 ¸i

:3

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13

15

20

19

16

12

17

21

18

22

XX

MA Figure 1

Figure 2

Representative karyotype from case 1. Karyotype is described in Table 2.

Representative karyotype from the metastases of fase 6. Karyotype is described in Table 2.

!

2

6

13

15

9

16

10

1!

17

12!

181

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19

MARKERS

8

14

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~

3

20

Ii I

21

22

XI

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Y

Study of Primary Squamous Cell Carcinomas

57

6 20

Figure 3

Representative karyotype from case 19. Karyotype is described in Table 2.

Figure 4

Representative karyotype from case 53. Karyotype is described in Table 2.

X

58

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12.

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