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Expression and CpG Methylation of the Insulin-like Growth Factor II Gene in Human Smooth Muscle Tumors¹

Laboratory for Physiological Chemistry; University of Utrecht, Vondellaan 24A, NL-3521 GG Utrecht, the Netherlands [T. G., I. P., J. S. S.]; Departments of Pathology [L. T. M. V. D. V., W. D. O.] and Internal Medicine [C. J. M. L.], Academic Hospital Utrecht, NL-3508 GA Utrecht, the Netherlands; and INSERM Unité 142, Hopital Saint , Antoine, 75571 Paris Cedex 12, France [H. S.]

² To whom requests for reprints should be addressed.

Previously we have shown that expression of the insulin-like growth factor II (IGF-II) gene in 36 normal smooth muscle tissues (myometria) and 26 benign smooth muscle tumors (leiomyomas) was detectable by Northern blot analysis but that the RNA levels were low. In 9 of 20 malignant smooth muscle tumors (leiomyosarcomas) IGF-II gene expression was also low or absent, while in 11 of 20 the IGF-II gene was abundantly expressed.

In 32 of these tissues we have now studied the DNA methylation state of the IGF-II gene. For the analysis of overall methylation of the gene the restriction endonucleases HpaII and MspI were used. In normal smooth muscle and in leiomyomas the IGF-II gene appeared to be methylated. In leiomyosarcomas with low IGF-II gene expression the DNA was partly demethylated. In leiomyosarcomas with abundant IGF-II gene expression overall methylation of the DNA tended to be low. In addition, we have studied the methylation state of one particular CpG site in the IGF-II gene with the restriction endonuclease AvaII. The results of the latter analysis confirm the analysis with HpaII and MspI.

In conclusion, in malignant smooth muscle tumors the data indicate an inverse correlation between CpG methylation and expression of the IGF-II gene.

¹ Supported by Grant UUKC 88-04 from the Netherlands Cancer Foundation (Nederlandse Kankerbestrijding).

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Received 6/22/92. Accepted 9/23/92.