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[Cancer Research 55, 2266-2269, June 1, 1995]
© 1995 American Association for Cancer Research
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Molecular Analysis of the Cyclin-dependent Kinase Inhibitor Gene p27/Kip1 in Human Malignancies1

Norihiko Kawamata2, Roberta Morosetti, Carl W. Miller, Dorothy Park, Konstantin S. Spirin, Tsuyoshi Nakamaki, Seisho Takeuchi, Yoshihiro Hatta, Jean Simpson, Sharon Wilczynski, Young Yiul Lee, Claus R. Bartram and H. Phillip Koeffler

Division of Hematology/Oncology, Cedars-Sinai Research Institute, UCLA School of Medicine, Los Angeles, California 90048 [N. K., R. M., C. W. M., D. P., K. S. S., T. N., S. T., Y. H., H. P. K.]; Department of Pathology, City of Hope National Medical Center, Duarte, California 91010 [J. S., S. W.]; Division of Hematology/Oncology, Department of Internal Medicine, Han Yang University Hospital, Sung Dong-ku, Seoul 133-792, Korea [Y. Y. L.]; and Section of Molecular Biology, Department of Pediatrics II, University of Ulm, Ulm, Germany [C. R. B.]

Cyclin and cyclin-dependent kinase (CDK) complexes play important roles in controlling the cell cycle. The CDK inhibitors (CDKIs) inhibit the kinase activities of the complexes and block transitions of the cell cycle. Recently several CDKI genes have been cloned, and evidence suggests that at least a couple of these may be tumor suppressor genes. In this study, the partial structure of a CDKI gene, p27/Kip1, was determined. In addition, a large number of human cancers (432 cases) and cancer cell lines (20 lines) were analyzed for alterations of the p27/Kip1 gene by Southern blot analysis and PCR/single-strand conformation polymorphism. The coding region of the p27/Kip1 gene consists of at least two exons and an intron of about 600 bp. In 140 tumors of various tissues and 18 transformed cell lines, no deletions or rearrangements of the gene were detected by Southern blot analysis using a part of the coding sequence as a probe. One polymorphism and one silent mutation were detected by PCR/single-strand conformation polymoprhism. The polymorphism was a nucleotide substitution of guanine for thymine (GTC->GGC) at codon 109, resulting in an amino acid substitution of glycine for valine (Val->Gly). In summary, no abnormalities of the p27/Kip1 gene were detected in human malignancies. Now, two groups of CDKIs are classified based on the structure of the proteins. One group includes the p15, p16, and p18 CDKIs, which have ankyrin repeat motifs. The p15 and p16 CDKI genes are very frequently mutated in a variety of cancers. The p27/Kip1 and p21 CDKIs belong to the other group. We reported previously that abnormalities of the p21 gene were very rare. The latter group of the CDKIs, including p27/Kip1 and p21, are rarely mutated in human malignancies.

1 This work was supported in part by a grant from The Ryoichi Naito Foundation for Medical Research (to N. K.) as well as NIH Grants DK42792, CA42710, and CA26038, the Concern Foundation, and the Parker Hughes Trust.

2 To whom requests for reprints should be addressed, at Cedars-Sinai Medical Institute, UCLA School of Medicine, 8700 Beverly Blvd., D-5066, Los Angeles, CA 90048.

Received 3/ 8/95. Accepted 3/31/95.




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