ABSTRACT
OBJECTIVE: To determine the polymorphisms in the 3' untranslated region (3'UTR) of the G6PD gene and analyze the specific SNPs or haplotypes that may affect messenger RNA (mRNA) secondary structure by in silico analysis. METHODS: We studied the 3'UTR polymorphisms in 107 healthy subjects living in Thailand (ethnicities unknown). The haplotype was analyzed using Haploview software. mRNA secondary structure and microRNA binding of the G6PD gene were predicted by use of the CLC Main Workbench 6.9 and RegRNA 2.0 programs. RESULTS: The results revealed 1 new variant in the 3'UTR of the G6PD gene (c.*99A > G). Haplotype ATCG showed significance by association-test results that yielded a high odds ratio (OR, 7.90; 95% confidence interval [CI], 3.15-19.81). Moreover, this haplotype affected G6PD mRNA secondary structure changes and microRNA binding via in silico analysis. CONCLUSIONS: These results suggest that the haplotype ATCG is associated with reduction of G6PD enzyme expression, G6PD mRNA secondary structure changes, and microRNA binding via in silico analysis.
Subject(s)
3' Untranslated Regions/genetics , Glucosephosphate Dehydrogenase Deficiency/genetics , Glucosephosphate Dehydrogenase , Haplotypes/genetics , Polymorphism, Single Nucleotide/genetics , Adult , Female , Glucosephosphate Dehydrogenase/chemistry , Glucosephosphate Dehydrogenase/genetics , Glucosephosphate Dehydrogenase/ultrastructure , Humans , Linkage Disequilibrium , Male , Middle Aged , Nucleic Acid Conformation , RNA, Messenger/genetics , Young AdultABSTRACT
In response to oxidative stress and during apoptosis, cells often shed microparticles (MPs), submicron elements carrying phosphatidylserine and protein antigens. Glucose-6-phosphate dehydrogenase (G6PD)-deficient cells are extremely sensitive to oxidative damage that may lead to the formation of MPs. To determine whether G6PD deficiency alters membrane phospholipid asymmetry and increases MPs production, we determined the concentrations and cellular origins of MPs in G6PD-deficient individuals using flow cytometry. G6PD-deficient individuals showed an increase in circulating MPs concentrations as compared with G6PD-normal individuals [1051/µL (865-2532/µL) vs. 258/µL (235-575/µL), P < 0.01]. MPs concentrations were significantly increased with the severity of G6PD deficiency. Median MPs concentrations from individuals with severe G6PD deficiency, and individuals with moderate G6PD deficiency were 2567/µL (1216-2532/µL) and 984/µL (685-2107/µL), respectively (P < 0.01). Importantly, G6PD enzymatic activity was significantly correlated with MPs concentrations with r(2) = 0.731. MPs found in G6PD deficiency individuals were largely derived from red blood cells (RBCs) (45%) and platelets (30%). Additionally, Atomic Force Microscopy was used to study the morphology and measures the diameter of MPs found in G6PD-deficient individuals. The mean (SD) width and height of RMPs were 0. 41 (0.18) and 2.04 (0.14) µm, respectively. Together, these results indicate that MP concentration is significantly correlated with G6PD enzymatic activity and is increased in G6PD-deficient as compared with G6PD-normal individuals. Our data also provide an evidence for an alteration in cell membrane associated with a decreased in G6PD activity. However, the significance of MPs in G6PD deficiency needs further clarification.
