Expression of Human Placental-Type 6-Phosphofructo-2-kinase/Fructose 2,6-Bisphosphatase in Various Cells and Cell Lines

Abstract

The expression of the human placental-type 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (HP2K) in various human cells and cell lines was investigated at the levels of transcription and translation. Analyses by both Northern blotting and a reverse transcription-polymerase chain reaction (RT-PCR) showed that BeWo, U-937, SupT1, H9, HeLa, HepG2, and human mononuclear cells, as well as human placental chorionic cells, expressed HP2K mRNA. All the nucleotide sequences of RT-PCR products from these cell lines were identical to that of HP2K. The expression of HP2K protein was determined by Western blot analysis of fractions from POROS-HQ column chromatography of the cell extracts from U-937 cells, which was used as an example of HP2K-mRNA positive cell lines. As with the 6-phosphofructo-2-kinase activity of HP2K, the activity of 6-phosphofructo-2-kinase in extracts of U-937 cells was not inhibited by glycerol 3-phosphate, a known 6-phosphofructo-2-kinase inhibitor of liver- and testis-type isozymes. These results strongly suggested that various cell lines, in particular U-937 cells, express functional HP2K enzyme. Furthermore, 6-phosphofructo-2-kinase in U-937 cells was found to be activated by treatments with isoproterenol and phorbol 12-myristate 13-acetate, indicating regulation of 6-phosphofructo-2-kinase activity in U-937 cells by protein kinases A and C.