Cytogenetic Analysis of an Adenoid Cystic Carcinoma of the Bartholin's Gland A Rare, Semimalignant Tumor of the Female Genitourinary Tract Marion Kiechle-Schwarz, Friedrich Kommoss, Johanne Schmidt, Lilian Lukovic, Lioba Walz, Thomas Bauknecht, and Albrecht Pfleiderer

ABSTRACT: Cytogenetic analysis has been performed on short-term cultures from a 56-year-old woman suffering from an adenoid cystic carcinoma of Bartholin' s gland. Beside a normal female karyotype, the t u m o r revealed an abnormal cell line with complex chromosome changes involving the chromosomes 1, 4, 6, 11, 22, and 14. The mainly structural and nonbalanced rearrangements led to the loss of the chromosome segments 1p31-*qter, 4q22--~q28, 6p12-~qter, 11p11.2-~pter, 14q24-~qter, and 22q13--~qter. Clonal numerical aberrations were not observed. To our knowledge, such a t u m o r has todate not been cytogenetically investigated.

INTRODUCTION

CASE PRESENTATION

Primary a d e n o i d cystic carcinoma of Bartholin's gland is a rare neoplasm, and only 37 cases have been reported in the literature [1]. The t u m o r comprises about 2 0 - 2 5 % of all p r i m a r y carcinomas of Bartholin's gland according to the largest series p u b l i s h e d to date [2]. A l t h o u g h a d e n o i d cystic carcinoma has been suspected to have its origin in glandular or duct e p i t h e l i u m [3], or in m y o e p i t h e l i a l cells [4], histogenesis of this unusual neoplasm, w h i c h more c o m m o n l y involves the salivary glands, oral cavity, nasopharynx, breast, and skin, remains unsettled [5]. The tumor u s u a l l y shows a protracted clinical course, and it tends to r e p e a t e d l y recur locally. Metastases to regional l y m p h nodes, lungs, and bones can occur, and patients m a y live for e x t e n d e d intervals even with metastatic disease. W h i l e only 39% of patients are progression free 10 years after diagnosis, the 1O-year survival rate of all patients with sufficient follow up in the literature is 59%

A 56-year-old white female with a n o n c o n t r i b u t o r y gynecologic history first presented with a painful p a l p a b l e vulvar mass at the age of 34. In 1969 an a d e n o i d cystic c a r c i n o m a of the Bartholin's gland had been r e m o v e d from the left vaginal wall. Subsequently, the patient u n d e r w e n t surgical therapy almost a n n u a l l y due to local recurrence of the tumor. In the years 1976, 1977, 1981, 1983, 1987 and 1989, local recurrent tumors had been r e m o v e d from the vulva, the perineum, the periurethral region, and the fossa ischiorectalis. In January 1991, the 11th local recurrence occurred affecting the vagina, anus, and s e p t u m rectovaginale with infiltration of the rectum. Radical surgical treatment consisted of a partial vaginectomy and a b d o m i n o p e r i n e a l rectum resection with a terminal colostomy. F o l l o w - u p examinations 6 and 12 months later s h o w e d no evidence of disease. Grossly, the t u m o r consisted of an ill-defined, fibrous infiltrating mass measuring 8 x 5 x 5 cm with a hard white cut surface. Histologically, the t u m o r was c o m p o s e d of nests of fairly uniform small basaloid cells w i t h hyperchromatic nuclei and scant slightly b a s o p h i l i c cytoplasm, arranged in a cribriform pattern. The p s e u d o c y s t i c spaces were filled with acellular material in a typical fashion [6] (Fig. 1).

[1]. S i n c e cytogenetic studies on a d e n o i d cystic carcinomas of the Bartholin's gland are virtually nonexistent, we w o u l d like to c o m m u n i c a t e the c h r o m o s o m a l findings in such a tumor from a 56-year-old w o m a n and to discuss the results in the light of previous reports on karyotypic abnormalities in benign and malignant tumors.

From the Department of Gynecology and Obstetrics, AlbertLudwigs University, Freiburg, Germany. Address reprint requests to: Dr. Marion Kiechle-Schwarz, Universit~ts-Frauenklinik, Hugstetterstrasse 55, 7800 Freiburg i. Br., Germany. Received December 19, 1991; accepted January 22, 1992. 26 Cancer Genet Cytogenet 61:26-30 (1992) 0165-4608/92/$05.00

CYTOGENETIC METHODS AND RESULTS Cytogenetic analysis was carried out on short-term cultures from fresh tumor samples of the a d e n o i d cystic c a r c i n o m a obtained at surgery. Recurrent t u m o r n o d u l i were r e m o v e d from the vagina, perianal region, and rectum. Tissue pieces © 1992 Elsevier Science Publishing Co., Inc. 655 Avenue of the Americas, New York, NY 10010

Cytogenetics of an Adenoid Cystic Carcinoma of the Bartholin's Gland

27

Figure I Adenoid cystic carcinoma of the Bartholin's gland showing the cribriform growth pattern. Pseudocystic spaces filled with acellular material. Hematoxylin & eosin, x 400.

were minced with scalpels and disaggregated for 2 hours with collagenase II (GIBCO) at a final concentration of 200 U/ml. Single-cell suspensions were initiated in flasks and on glass chamber slides using RPMI 1640 medium supplemented with fetal calf serum (17%), L-glutamine (200 mM), penicillin (5000 U/ml), streptomycin (5000 mg/ml), insulin (0.01 mg/ml), EGF (2 ng/ml), and cholera toxin (100 ng/ml) at 37°C and 5% CO2. Cells were harvested after 5-10 days of growth. Air-dried chromosome preparations were obtained according to standard procedures, including overnight Colcemid treatment (0.01/xg/ml) and hypotonic shock (0.4% KC1) for 30 min and subsequent fixation in methanol/acetic acid (3 : 1 v/v) for 3 x 20 min. For the chromosome preparation from the cells grown on glass chamber slides, 0.3% NaC1 was used as a hypotonic solution. Metaphases were G-banded using trypsin Giemsa [7]. Karyotypes were constructed according to ISCN (1985) [8]. A total of 30 metaphases obtained from short-term cultures of the adenoid cystic carcinoma of the vagina, perianal region, and rectum were analyzed and displayed the following karyotypes: 46,XX 45,XX,- 1,del(4)(q22q28),- 6,der(11)t(6;11) (p12;p 11.2),der(14)t(14;?)(q24;?), der(22)t(1;22)(p31;q13), -4-marker

[17 cells].

[13 cells].

In five cells additional nonclonal numerical changes were found. A complete G-banded karyotype is shown in Figure 2. Partial karyotypes showing the aberrant chromosomes 4, 6, 11, 14, 22, and the marker in detail are illustrated in Figure 3. DISCUSSION

A review of the cytogenetic literature showed that this is the first report on chromosomal findings in adenoid cystic carcinoma of the Bartholin's gland. Beside a normal female

cell line, an aberrant clone with a pseudodiploid modal chromosome number was observed. The structural rearrangements affected chromosomes 1, 4, 6, 11, 14, and 22 and led to the loss of the segments 1p31--~qter, 4q22--~q28, 6p12--~qter, 11pll.2--~pter, 14q24-~qter, and 22q13-~qter. One metacentric marker chromosome which is most probably composed of same of these segments additionally appeared in the karyotypically abnormal tumor cells. However, there is still a net loss of chromosomal material in this tumor, since the marker chromosome has the size of an E-group chromosome. Most of the rearrangements affected bands or regions which are recurrently involved in chromosome aberrations observed in other benign or malignant solid tumors. Chromosome I alterations are the most common cytogenetic findings in human malignancies [9]. They have been found in many solid tumors of various origin and in hematological disorders, and are regarded to represent late, secondary, and rather nonspecific changes. Especially, trisomy of the long arm of chromosome I appeared to be associated with tumor progression. Monosomy for the long arm of chromosome 1, however, has not yet been consistently found in a special type of tumor. In this case, it could be speculated that the low potential to metastasize is due to the underrepresentation of genes located on the long arm of chromosome 1. The rearrangements of chromosome 4 (interstitial deletion of 4q) and the loss of chromosome 6 or changes affecting band 6p12 have not been recurrently found in solid tumors [10]. Deletions of the short arm of chromosome 11, however, have been consistently found in certain kinds of malignancies, e.g., Wilms' tumor and bladder cancer [11]. In ovarian cancer there is cytogenetic and molecular evidence that a putative tumor suppressor gene which is localized in this chromosome region plays an important role in ovarian cancer development [12, 13].

F

__

19

~3

6

Figure 2

20

- ~t

-I

MARKERS

9

21

16

10

!---

22

17

11

4

Representative G-banded karyotype. Abnormal chromosomes are indicated by arrowheads.

~-

~ ~--!

~~ 15

8

7

14

3

2

-I

X

18

12

5

Y

-I

Cytogenetics of an Adenoid Cystic Carcinoma of the Bartholin's Gland

29

4/del(4q)

6/t(6;ii)/ii

14/der (14)

22/der(22)

--m--,-t. M Figure 3 Representative G-banded partial karyotypes illustrating two examples of each rearrangement.

Interestingly, the band 14q24, which is consistently rearranged in leiomyomas of the uterus [14], the most common benign gynecological tumor, is also affected in this adenoid cystic carcinoma. This finding might point to the possible myoepithelial origin of this tumor. Regarding the alterations of chromosome 22 in this tumor, it is well known that deletions of 22q12-qter or monosomy 22 are specific findings in meningiomas [15], but were recurrently observed in leiomyomas, too [16]. The consistent abnormalities described showed the clonal origin of this tumor. It is obvious that we need more cytogenetic investigations of adenoid cystic carcinomas to identify recurrent, specific changes in these tumors. They

might help to clarify the origin of adenoid cystic carcinomas. It has been shown for a variety of tumors that cytogenetic aberrations may define certain pathological and histiogenetic entities [16, 17]. Moreover, the cytogenetic study of semimalignant tumors with a less aggressive clinical behavior might help to identify chromosome regions which are responsible for tumor progression and/ or metastasis.

This work has been supported by the German Research Council, Deutsche Forschungsgemeinschaft, AZ Ki 352/2-1.

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