PMID: 11309345Apr 20, 2001Paper

Folylpoly-gamma-glutamate synthetase gene mRNA splice variants and protein expression in primary human leukemia cells, cell lines, and normal human tissues

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research
G J Leclerc, J C Barredo

Abstract

Folypoly-gamma-glutamate synthetase (FPGS) is essential for the cytotoxicity of "classical" antifolates and their efficacy in cancer chemotherapy. The expression of the FPGS gene appears controlled by both tissue/lineage-specific and proliferation-dependent mechanisms. Four alternative exon 1 splice variants of the FPGS gene have been described previously, but their significance in gene regulation has not been determined. Furthermore, alternative splicing of the human FPGS gene in normal or transformed cells in vivo has not been reported. We have examined the mRNA expression of these FPGS splice variants in primary human leukemia cells, cell lines, and normal human hematopoietic progenitors using reverse transcription-PCR. Specific primers were designed to amplify splice variants 1, 1A, 1B, and 1C; and full-length FPGS mRNA was amplified using primers for exons 14 and 15 at the 3' end of the gene. In this study, we demonstrate that all four alternative exon 1 variants are expressed in all primary leukemia cells and cell lines (ALL and AML), as well as in normal human hematopoietic progenitors. No significant differences in mRNA expression were detected in primary cells or cell lines for the four exon 1 splice variants. Normal c...Continue Reading

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