Loss of normal allele of the APC gene in an adrenocortical carcinoma from a patient with familial adenomatous polyposis
Madoka Seki1, Kiyoko Tanaka1, Rei Kikuchi-Yanoshita1, Motoko Konishi1, Hiroyuki Fukunari2, Takeo Iwama2, and Michiko Miyaki1
1 Department of Biochemsitry, The Tokyo Metropolitan Institute of Medical Science, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo, 113 Japan
2 Department of Surgery, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, Tokyo, 113 Japan
Received September 1, 1991 / Revised December 5, 1991
Summary. Endocrine neoplasms have been reported oc- casionally in patients with familial adenomatous poly- posis (FAP). An adrenocorotical carcinoma was studied in a patient with a family history of FAP. Loss of hetero- zygosity (LOH) in the region close to the adenomatous polyposis coli (APC) gene was detected in this carcinoma, and evidence was obtained that there was a loss of the normal allele of the APC gene. This is the first demon- stration of LOH at the APC locus in adrenocortical tu- mors. The present results and our previous data on LOH in a recurring desmoid tumor suggest that the heterozyg- ous mutant/wild-type condition of the APC gene may give rise to benign tumors, and that functional loss of this gene leads to development of tumors not only in the colon but also in other various parts of the body in FAP patients.
Introduction
Familial adenomatous polyposis (FAP) is an autosomal dominant trait that is determined by the heterozygous mutant APC gene on chromosome 5q21-22 (Herrera et al. 1986; Bodmer et al. 1987; Leppert et al. 1987) and characterized by numerous benign colorectal adenomas and a high incidence of colorectal carcinomas. FAP is not solely colonic disease, and there have been reports of other benign and malignant tumors occurring in pa- tients, including gastric and duodenal polyps and car- cinomas, osteomas, desmoid tumors and endocrine ade- nomas and carcinomas (Jagelman 1987). It has also been reported that endocrine adenoma is significantly more frequent in FAP than in other cancer patients (Schneider et al. 1983; Painter et al. 1985). These data suggest that the APC gene may play an important role in growth dis- order and tumroigenesis in various parts of the body in FAP patients.
We have previously observed the loss of the normal allele of the APC gene not only in many colonic tumors from FAP patients (Miyaki et al. 1990) but also in a de- smoid tumor from a patient (Okamoto et al. 1990). To examine the pleiotropic effect of inactivation of the APC gene, we performed restriction fragment length poly- morphism (RFLP) analysis on an adrenocortical carcino- ma formed in an FAP patient and detected loss of het- erozygosity (LOH) in the APC region.
Materials and methods
Tumor and normal tissues were obtained from FAP patients treated at Tokyo Medical and Dental University. A tumor sample from PLK91 (63-year-old woman) was diagnosed histopathologically as adrenocortical carcinoma. DNA extraction, Southern blotting and hybridization were performed as described previously (Miyaki et al. 1990). The following RFLP probes were used: L-myc (1p32), YNZ2 (1p), D3S2 (3p14-21), D5S6 (5q13), D5S37 (5q21-22), D5S71 (5q21-22), D5S81 (5q21-22), H-ras-I (11p15.5), INS (11p15), D11S12 (11p15.5), INT2 (11q13), D17S30 (17p13.3), D18S5 (18q21.3-ter) and IGLC (22q11.1-11.2).
Results and discussion
An adrenocortical carcinoma was found in an FAP pa- tient (PLK91). To elucidate the contribution of the APC gene in this tumor we analyzed LOH in the region close to the APC gene. Two probes, D5S81 (YN5.48) and D5S37 (Pi227) which have been reported to be closely linked to APC (Nakamura et al. 1988; Stewart et al. 1987; Meera Khan et al. 1988) detected the LOH in this tumor as indicated in Fig. 1. These probes detected fre- quent LOH in colorectal tumors from FAP patients (Mi- yaki et al. 1990) and also loss in a desmoid tumor (Oka- moto et al. 1990). To examine which allele of the pair at the APC locus was lost in the tumor, we performed RFLP analysis on normal tissues in the patient’s brother, daugh-
D5S81 / Mspl
D5S37 /Pst |
N Ca
N Ca
a1
a1
a3
a2
PLK91
PLK91
ter and grandchild with a history of FAP (Fig. 2). Nor- mal tissue from PLK91 was heterozygous in D5S81 and D5S37, and allele 1 of D5S81 and allele 1 of D5S37 were absent in her adrenocortical carcinoma (Ca). Allele 3 of D5S81 and allele 2 of D5S37 were retained. These alleles were inherited by her affected brother (D16), affected daughter (PLK61) and affected grandchild (D45), and the mutant allele of the APC gene was assumed to be re- tained with these alleles in this tumor. This result pro- vided evidence for loss of the normal allele of the APC gene, which leads to loss of the suppressing function of the APC gene in the adrenocortical carcinoma.
LOH on chromosomes 1, 3, 11, 17, 18 and 22 was also examined. However, no LOH was detected in H-ras-1, D11S12 and INS on chromosome 11p which have been reported to be lacking in adrenocortical carcinomas from patients with Beckwith-Wiedemann (BW) syndrome and familial adrenocortical carcinoma (ADCC) (Henry et al. 1989) and in malignant adrenocortical carcinomas from sporadic cases (Yano et al. 1989). The genes for the he- reditary diseases, BW and ADCC, are located in this re- gion. The reason for the absence of LOH on chromosome
11p in the same tumor in this FAP patient may be inter- preted as follows: (1) the probes used in the present study could not detect LOH because they are located far from the BW or ADCC gene; (2) both alleles of BW or ADCC were inactivated by point mutations; or (3) in the case of FAP patients, inactivation of the APC gene contributes to development of adrenocortical tumors instead of inac- tivation of the gene on chromosome 11p.
It has been suggested from our previous results that the heterozygous mutant/wild-type condition of the APC gene may bring about the formation of benign colorectal adenomas with mild or moderate dysplasia, and loss of the normal allele of the APC gene develops adenomas to a condition of severe dysplasia. The severe adenomas may then convert and progress into malignant carcinomas accompanied by LOH in tumor suppressor genes on chro- mosomes 17p, 18q, 22q, etc. Similar inactivation of the APC gene has been suggested to contribute to the de- velopment of extracolonic tumors in FAP patients by the fact that LOH of the APC region was detected in a large recurring desmoid tumor, but has not been detected in small desmoid tumors. The LOH in a malignant adreno- cortical tumor described in this report may support the idea that half loss of the tumor suppressor activity of the APC gene is the cause of extracolonic benign tumors in FAP patients, and that full loss causes development of the tumors to an advanced stage. Further study with a large number of tumors is needed to confirm this hypo- thesis, and more detailed analysis of the genetic changes of the APC gene in tumors is also required, since the structure of the APC gene has recently been clarified (Groden et al. 1991; Kinzler et al. 1991).
Acknowledgements. This work was supported in part by Grants-in- Aid for Cancer Research from the Ministry of Education, Science and Culture of Japan. We are grateful to Dr. Y. Nakamura, Dr. G. D. Stewart and other scientists for providing YN5.48, Pi227 and other probes. Several probes were provided through the Japanese Cancer Research Resources Bank.
D16
N
D5S37 2
2
D5S71 2
2
PLK91
N
Ca
APC
+
*
D5S37
1
2
2
D5S81 1
3
D5S71
2
2
2
APC
+
*
*
D5S81
1
3
3
PLK61
N
D5S37
1
2
D5S71
2
2
APC
+ *
D5S81
3
3
D45
N
D5S37
2
2
D5S71 2
2
APC + *
D5S81 2 3
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