TRANSCRIPTIONAL REGULATION OF THE HUMAN PLACENTAL-LIKE ALKALINE-PHOSPHATASE GENE AND MECHANISMS INVOLVED IN ITS INDUCTION BY SODIUM-BUTYRATE

The human alkaline phosphatases constitute a multigene family with at least four members. Placental-like alkaline phosphatase (PLAP) is of particular interest because it is frequently present in tumors, where it serves as a marker of malignant transformation. Moreover, its expression is highly inducible by differentiating agents such as sodium butyrate. In the present study we have examined the PLAP gene promoter in order to better understand the mechanisms involved in its expression and induction. The PLAP promoters from four colon cancer cell lines with widely varied butyrate-inducible alkaline phosphatase activity were thermally amplified and sequenced. The overall sequence similarity of this region was found to be 99% between cell lines; thus, sequence variation of the promoter does not appear to account for the differential expression of this marker. We therefore analyzed the activity of the LS174T cell PLAP promoter using transient transfection experiments. Here, the 5'-flanking region of the gene was found to have positive regulatory elements in nucleotides -1 to -170 and -363 to -512 (relative to the start of transcription). A negative control element was also found to be present in the region between nucleotides -170 and -363. Mobility shift electrophoresis indicated that a nuclear factor bound to the promoter between bases -182 and -341. Furthermore, the activity of the PLAP promoter was found to be inducible by sodium butyrate. In contrast, the closely related placental alkaline phosphatase gene promoter exhibited almost no response to this agent. These results confirm that the activity of the PLAP promoter is stimulated by sodium butyrate and delineate regions that control this induction process.