RRC ID 39192
Author Murakami Y, Nobukuni T, Tamura K, Maruyama T, Sekiya T, Arai Y, Gomyou H, Tanigami A, Ohki M, Cabin D, Frischmeyer P, Hunt P, Reeves RH.
Title Localization of tumor suppressor activity important in nonsmall cell lung carcinoma on chromosome 11q.
Journal Proc Natl Acad Sci U S A
Abstract Loss of heterozygosity on chromosome 11q23 is observed at high frequency in human nonsmall cell lung carcinomas (NSCLCs), suggesting the presence of a tumor suppressor gene. Previous analysis of DNA from 79 patients identified a commonly deleted segment of 5 centimorgans. Complementation analysis was used to further localize a putative tumor suppressor gene. Three yeast artificial chromosome (YAC) clones spanning the minimal loss of heterozygosity region were modified, and spheroplast fusion was used to transfer them into human A549 NSCLC or murine Lewis lung carcinoma (LLC) cell lines. The resulting yeast x human hybrid cell lines containing an intact copy of a 1.6-Mb YAC, 939b12, showed reduced growth in vitro. Injection of parental A549 cells into athymic (nu/nu) mice resulted in tumor formation at 27 of 28 injection sites. In contrast, two independent 939b12-containing cell lines formed tumors at only 3 of 20 injection sites. 939b12 also suppressed tumor formation by LLC NSCLC cells in nude mice, but YACs 785e12 and 911f2, which flank 939b12, had no suppressor activity. Further localization of tumor suppression activity on 939b12 was accomplished by introduction of defined fragmentation derivatives into A549 cells and by analysis of YACs that were broken on transfer into LLC cells. This complementation approach localized tumor suppression activity to the central 700 kb of 939b12 and provides a functional assay for positional cloning of this tumor suppressor gene.
Volume 95(14)
Pages 8153-8
Published 1998-7-7
DOI 10.1073/pnas.95.14.8153
PMID 9653156
PMC PMC20945
MeSH Animals Carcinoma, Non-Small-Cell Lung / genetics* Chromosome Mapping Chromosomes, Artificial, Yeast Chromosomes, Human, Pair 11* Genes, Tumor Suppressor* Humans Lung Neoplasms / genetics* Mice Mice, Nude Neoplasms, Experimental / genetics Neoplasms, Experimental / pathology
IF 9.58
Times Cited 74
Human and Animal Cells