Modulation of iron on mitochondrial aconitase expression in human prostatic carcinoma cells.
The mitochondrial aconitase (mACON) containing a [4Fe-4S] cluster is regarded as the key enzyme for citrate oxidation in the epithelial cells of human prostate. In vitro studies using the human prostatic carcinoma cells, PC-3 cells, found that both hemin and ferric ammonium citrate (FAC) significantly increased mACON enzymatic activity and gene expression. The effect of FAC on mACON was enhanced 2-fold by co-treating with ascorbic acid but blocked by co-treating with iron chelator, deferoxamine mesylate. Hemin treatments blocked 30% of citrate secretion from PC-3 cells but upregualted 2-fold of intracellular ATP biosynthesis. Results from reporter assay by using a cytomegalovirus enhance/promoter driven luciferase mRNA ligated to the iron response element (IRE) of mACON as a reporter construct demonstrated that modulation of FAC on gene translation of mACON gene is dependent on the IRE. Transient gene expression assays indicated that upregulation of mACON gene transcription by FAC may through the putative antioxidant response element (ARE) signal pathway. This study provides the first evidence of the biologic mechanism of human mACON gene translation/transcription and suggests a regulatory link between the energy utilization and the iron metabolism in human prostatic carcinoma cells.
Juang
Horng-Heng
HH
Department of Anatomy, Chang Gung University, Tao-Yuan 333, Taiwan, ROC. hhj143@mail.cgu.edu.tw
D000154:Aconitate Hydratase / Q000096:biosynthesis*; D000255:Adenosine Triphosphate / Q000378:metabolism; D001205:Ascorbic Acid / Q000494:pharmacology; D001483:Base Sequence; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002614:Chelating Agents / Q000494:pharmacology; D002951:Citrates / Q000378:metabolism; D003676:Deferoxamine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005290:Ferric Compounds / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D005822:Genetic Vectors; D006427:Hemin / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D007501:Iron / Q000378:metabolism*; D008297:Male; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011471:Prostatic Neoplasms / Q000201:enzymology*; D014176:Protein Biosynthesis; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D015398:Signal Transduction; D014158:Transcription, Genetic; D015854:Up-Regulation
D002614:Chelating Agents; D002951:Citrates; D005290:Ferric Compounds; D000644:Quaternary Ammonium Compounds; D006427:Hemin; D000255:Adenosine Triphosphate; D007501:Iron; D000154:Aconitate Hydratase; D003676:Deferoxamine; D001205:Ascorbic Acid; C013531:ferric ammonium citrate