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Defining regulatory elements in the human KAI1 (CD 82) metastasis suppressor gene

The human KAI1 metastasis suppressor gene encodes for a 267 amino acid plasma membrane glycoprotein, which has four transmembrane domains and one large and one small extracellular domain. Plasma membrane expression of KAI1 is downregulated during the progression of several cancers to a metastatic state, including prostate, lung, and pancreatic cancers. To elucidate the mechanisms for this downregulation, an understanding of its transcriptional regulation is critical. Therefore, a set of luciferase reporter gene plasmid constructs were generated containing various 5′ flanking regions to the transcription initiation site, with or without the first exon and a portion of the first intron of the KAI1 gene. These constructs were transfected into a series of normal and malignant prostate cell lines as well as fibroblasts and the resultant luciferase activities used to define the regulatory elements for expression of the KAI1 gene. Using this approach, the minimal KAI1 promoter was found to be 0.5 kb in size and contains two regions. The first region includes the 197 bp 5′ to the transcription initiation site and the second region includes the first exon and a portion of first intron (i.e., +1 to +351 bp). This combined region result in positively regulated strong, but not epithelial specific, luciferase expression. In addition to this minimal promoter element from −197 to +351 bp, there is a second regulatory element further upstream (i.e., −735 to −197 bp), which negatively regulates luciferase expression. Additional studies of Marreiros et al. (Gene 302:155, 2003) have documented that a third regulatory element even further 5′ (i.e., −922 to −846 bp) encodes an enhancer for the KAI1 gene. Prostate 57: 256–260, 2003. © 2003 Wiley‐Liss, Inc.