Individual expression features of GPX2, NQO1 and SQSTM1 transcript variants induced by hydrogen peroxide treatment in HeLa cells.

Pathway activity assessment-based approaches are becoming highly influential in various fields of biology and medicine. However, these approaches mostly rely on analysis of mRNA expression, and total mRNA from a given locus is measured in the majority of cases. Notably, a significant portion of protein-coding genes produces more than one transcript. This biological fact is responsible for significant noise when changes in total mRNA transcription of a single gene are analyzed. The NFE2L2/AP-1 pathway is an attractive target for biomedical applications. To date, there is a lack of data regarding the agreement in expression of even classical target genes of this pathway. In the present paper we analyzed whether transcript variants of GPX2, NQO1 and SQSTM1 were characterized by individual features of expression when HeLa cells were exposed to pro-oxidative stimulation with hydrogen peroxide. We found that all the transcripts (10 in total) appeared to be significantly individually regulated under the conditions tested. We conclude that individual transcripts, rather than total mRNA, are best markers of pathway activation. We also discuss here some biological roles of individual transcript regulation.

The Association of Adipokines and Myokines in the Blood of Obese Children and Adolescents with Lipoprotein Lipase rs328 Gene Variants.

Obesity develops largely due to genetic factors, with the genetic polymorphism of lipid metabolism enzymes being of particular importance. However, it is still unclear how the genetic variants of one of the key enzymes in lipid transport, lipoprotein lipase (LPL), are associated with the endocrine function of mesenchymal tissues in obesity. The current study was aimed at the investigation of the LPL rs328 gene variant association with adipokines and myokines levels, as well as lipid metabolism indices in the blood of children and adolescents of both genders with obesity. We found that LPL polymorphism rs328 is not characterized by the differences in the levels of hormones, adipokines, and myokines and in the blood of healthy children and adolescents; however, it significantly affects these indices during obesity in gender-dependent manner. The shifts in hormones, adipokines, and myokines manifest mostly in the obese individuals with Ser447Ser genotype rather than with 447Ter genotype. Obese boys homozygous for Ser447Ser have more elevated leptin levels than girls. They also demonstrate lower adiponectin, apelin, prolactin, and osteocrine levels than those in obese girls with the same genotype. The gender-based differences are less pronounced in individuals with 447Ter genotype than in the homozygotes for 447Ser. Thus, we conclude that the polymorphism rs328 of the lipoprotein lipase gene is accompanied by the changes in hormones, adipokines, and myokines levels in the blood of children and adolescents with obesity in gender-dependent manner.

Association of SNPs in Lipid Metabolism Gene Single Nucleotide Polymorphism with the Risk of Obesity in Children.

Background: Obesity is one of the most common metabolic disorders in the world, which develops due to an imbalance in energy consumption and expenditure, and both genetic and environmental factors are of great importance. We investigated the potential interactions of single nucleotide polymorphisms that might contribute to the development of polygenic obesity in children. Objective: The study involved 367 children and adolescents of both sexes aged from 4 to 18 years. The control group (normal weight) and the overweight groups included 65 and 302 children respectively. Methods: DNA for analysis was isolated from peripheral blood lymphocytes, then allelic variants rs99305069 of the FTO gene (chr16:53786615), Gln192Arg of the PON1 gene (chr7: 95308134), -250G>A of the LIPC gene (chr15: 58431740), and Ser447Ter of the LPL gene (chr8:19957678) were studied using the SNP-Express reagent kit. The results of allelic interactions were analyzed using the multifactor dimensionality reduction method. Results and Discussion: Among overweight children, the distribution of genotype and allele frequencies for the studied single nucleotide polymorphisms of the four genes corresponded to those of the control group (p > 0.05). It was found that in obese children SerSer homozygotes at the Ser447Ter polymorphism of the LPL gene, had serum triglyceride (TG) levels 2.3 times higher than in children with the same genotype from the control group. In overweight Ser447Ter heterozygotes (p < 0.0001), the TG level exceeded the control values by only 13% (p = 0.044). A two-locus genotype FTO AT/LPL SerTer, was associated with a reduced risk of childhood obesity.

Testing the concept of the interatomic status of the NFE2L2/AP1 pathway as a systemic biomarker for examination stress.

BACKGROUND AND OBJECTIVES: Oxidative status-based interactomic profiling is a promising approach for fundamental integrative cell biology, diagnostics, and therapy. However, this approach has been neither utilized as a method nor tested as a tool. Thus, we aimed (1) to develop an oxidative status pathway state assessment-based analytical procedure relying on NFE2L2/AP1 pathway evaluation, and (2) to preliminarily assess its responsiveness, performance and diagnostic properties when applied to deciphering stress conditions of the academic examination period and academic term. These conditions were chosen as those representing a common model of mild, everyday-life stressors causing shifts in oxidative status. METHODS: To meet the aim of the study, we performed a repetitive-measurements study collating gene expression of NFE2L2/AP1 pathway targets and controllers under the two stress conditions using semi-quantitative reverse transcription-polymerase chain reaction. RESULTS: Surprisingly, even with some sensitivity limitations of the methods employed, a pathway state analysis approach based on a multiple target-to-controller ratio calculation was highly responsive and yielded very high receiver operating characteristics in deciphering the model stress conditions. CONCLUSION: Although further testing of the approach is required, the interactomic pathway activation assaying concept was preliminarily experimentally proven to be a highly promising clinical diagnostic tool that may easily be adapted for current tasks.

Individual expression features of GPX2, NQO1 and SQSTM1 transcript variants induced by hydrogen peroxide treatment in HeLa cells

Abstract Pathway activity assessment-based approaches are becoming highly influential in various fields of biology and medicine. However, these approaches mostly rely on analysis of mRNA expression, and total mRNA from a given locus is measured in the majority of cases. Notably, a significant portion of protein-coding genes produces more than one transcript. This biological fact is responsible for significant noise when changes in total mRNA transcription of a single gene are analyzed. The NFE2L2/AP-1 pathway is an attractive target for biomedical applications. To date, there is a lack of data regarding the agreement in expression of even classical target genes of this pathway. In the present paper we analyzed whether transcript variants of GPX2, NQO1 and SQSTM1 were characterized by individual features of expression when HeLa cells were exposed to pro-oxidative stimulation with hydrogen peroxide. We found that all the transcripts (10 in total) appeared to be significantly individually regulated under the conditions tested. We conclude that individual transcripts, rather than total mRNA, are best markers of pathway activation. We also discuss here some biological roles of individual transcript regulation.