Manganese antagonizes iron blocking mitochondrial aconitase expression in human prostate carcinoma cells.
OBJECTIVE
To investigate the possible role of manganese in the regulation of mitochondrial aconitase (mACON) activity human prostate carcinoma cell line PC-3 cells.


METHODS
The mACON enzymatic activities of human prostate carcinoma cell line PC-3 cells were determined using a reduced nicotinamide adenine dinucleotide-coupled assay. Immunoblot and transient gene expression assays were used to study gene expression of the mACON. The putative response element for gene expression was identified using reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays.


RESULTS
In vitro study revealed that manganese chloride (MnCl2) treatment for 16 h inhibited the enzymatic activity of mACON, which induced the inhibition of citrate utility and cell proliferation of PC-3 cells. Although results from transient gene expression assays showed that MnCl2 treatment upregulated gene translation by approximately 5-fold through the iron response element pathway, immunoblot and reporter assays showed that MnCl2 treatments inhibited protein and gene expression of mACON. This effect was reversed by co-treatment with ferric ammonium citrate. Additional reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays suggested that a putative metal response element in the promoter of the mACON gene was involved in the regulation of MnCl2 on the gene expression of mACON.


CONCLUSIONS
These findings suggest that manganese acts as an antagonist of iron, disrupting the enzymatic activity and gene expression of mACON and citrate metabolism in the prostate.
Tsui
Ke-Hung
KH
Department of Anatomy, Chang Gung University, 259 Wen-Hua 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, China.
Chang
Phei-Lang
PL
Juang
Horng-Heng
HH
D000154:Aconitate Hydratase / Q000037:antagonists & inhibitors / Q000235:genetics*; D000199:Actins / Q000235:genetics; D000255:Adenosine Triphosphate / Q000378:metabolism; D045744:Cell Line, Tumor; D002712:Chlorides / Q000494:pharmacology*; D002951:Citrates / Q000378:metabolism; D017931:DNA Primers; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D017930:Genes, Reporter; D006801:Humans; D007501:Iron / Q000378:metabolism*; D008297:Male; D017895:Manganese Compounds / Q000494:pharmacology*; D008928:Mitochondria / Q000201:enzymology*; D011471:Prostatic Neoplasms / Q000201:enzymology
D000199:Actins; D002712:Chlorides; D002951:Citrates; D017931:DNA Primers; D017895:Manganese Compounds; D000255:Adenosine Triphosphate; D007501:Iron; D000154:Aconitate Hydratase; C025340:manganese chloride