Tag Archives: gene expression

Insulin resistance in obese adolescents and adult men modifies the expression of proliferation related genes

O. H. Minchenko1, Y. M. Viletska1, D. O. Minchenko1,2, V. V. Davydov3

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: ominchenko@yahoo.com;
2Bohomolets National Medical University, Kyiv, Ukraine
3SI “Institute of Children and Adolescent Health Care,
National Academy of Medical Sciences of Ukraine”, Kharkiv

Received: 11 December 2018; Accepted: 14 March 2019

Numerous data demonstrate that key regulatory factors, enzymes and receptors including HSPA5, MEST, SLC1A3, PDGFC, and ADM represent poly-functional, endoplasmic reticulum stress-dependent proteins, which control variable metabolic pathways. The expression level of genes of these proteins in the blood and subcutaneous adipose tissue of obese adolescents and adult men with and without insulin resistance was studied. It was shown that in blood of obese adolescents without insulin resistance the expression level of SLC1A3, HSPA5, MEST, and PDGFC genes was significantly increased, but development of insulin resis­tance led to down-regulation of these genes expression except HSPA5 gene as compared to the control group as well as to the group of obese adolescents without insulin resistance. At the same time, the expression level of ADM gene did not change significantly in obese adolescents without insulin resistance, but the development of insulin resistance led to down-regulation of this gene expression. In subcutaneous adipose tissue of obese adult men without insulin resistance the level of SLC1A3 gene expression was decreased, although ADM, MEST, and HSPA5 genes – increased. It was also shown that the development of insulin resistance in obese men affected the expression level of ADM and SLC1A3 genes only. Results of this investigation provide evidence that insulin resistance in obese adolescents and adult men is associated with specific changes in the expression of genes, which related to proliferation and development of obesity and insulin resistance as well as to endoplasmic reticulum stress and contribute to the development of obesity complications.

Folate-related processes in human placenta: gene expression, aminothiols, proliferation and apoptosis

M. Yu. Obolenskaya, R. R. Rodriges, O. P. Martsenyuk

Institute of Molecular Biology and Genetic, National Academy of Sciences of Ukraine, Kyiv;
e-mail: m.obolenska@gmail.com

The paper contains short information concerning the role of folate-related processes in cell metabolism and multiple diseases which are characterized by hyperhomocysteinemia. The authors represent more detailed information about the folate-related processes in human placenta, namely about the content of aminothiols at different allelic variants of placental methylenetetrahydrofolate reductase during the course of physiological pregnancy and preeclampsia. The existing data concerning the expression and catalytic activi­ty of corresponding enzymes are corroborated by the authors’ own results that proved for the first time the functional activity of transsulfuration pathway in human placenta. This pathway is activated in placental explants in parallel with down-regulation of proliferation and up-regulation of apoptosis when hyperhomocystei­nemia is imitated by high concentration of homocysteine in culture medium. On the whole the presented data point to the importance of placental folate-related processes for its normal function.

Gene expression of H(+)-pumps in plasma and vacuolar membranes of corn root cells under the effect of sodium ions and bioactive preparations

N. O. Kovalenko, T. A. Palladina

Kholodny Institute of Botany, National Academy of Sciences of Ukraine, Kyiv;
e-mail: tatiana_palladina@ukr.net

Four isoforms of H+-ATPase of plasma membrane: MHA1, MHA2, MHA3, MHA4 are expressed in the corn seedling roots with prevalence of genes MHA3 і MHA4. The exposure of seedlings in the presence of 0.1 M NaCl activated the expression of MHA4 gene isoform, that demonstrates its important role in the processes of adaptation to salinization conditions. In vacuolar membrane, where potential is created by two Н+-pumps, sodium ions activated gene expression of only Н+-АТРase of V-type, taking no effect on the expression of Н+-pyrophosphatase. The seeds pretreatment by synthetic preparations Methyure and Ivine did not affect gene expression of Н+-pumps. Thus we can suppose that the ability of the above preparations to activate functioning of Н+-pumps in the presence of sodium ions is realized at the post-tranlation level.

Expression of genes, encoding the enzymes of cysteine metabolism in human placenta in the first and third trimesters of uncomplicated pregnancy

K. L. Korneeva1, R. R. Rodriguez1, S. V. Ralchenko2, O. V. Martunovska2,
A. О. Frolova1, O. P. Martsenyuk1, L. V. Manzhula3, V. T. Melnyk4,
O. Y. Shkoropad4, M. Yu. Obolenska1

1Institute of Molecular Biology and Genetics, National Academy
of Sciences of Ukraine, Kyiv;
2Taras Shevchenko National University of Kyiv, Ukraine;
3Maternity Hospital of Kyiv N 3, Ukraine;
4Maternity Hospital of Irpin, Ukraine;
e-mail: m.obolenska@gmail.com

The cellular cysteine is highly regulated in a narrow range of concentrations due to its cyto- and neurtoxicity when it is overwhelmed or its deficiency for protein synthesis and other vital metabolic reactions when its amount is restricted. The regulation of cysteine content and its metabolic products, glutathione, taurine and inorganic sulfur compounds, is scarcely explored in human placenta though cysteine metabolism is closely related to the maintenance of redox status and protection from free radical oxidation, elimination of homocysteine and detoxification. These processes are particularly important for placenta which meets substantial changes of oxygen supply during its development, and is the last metabolically active organ between mother and fetus. The abundance of CDO, CSAD, ADO, SUOX, GCLC and GCLM mRNAs was estimated by RT-qPCR and compared with the computationally analyzed microarray gene expression data from GEO, while the level of individual protein – by western-blot analysis, both in placental samples from first and third trimesters of uncomplicated pregnancies. The abundance of CDO mRNA is significantly up-regulated at term compared to the first trimester, the level of GCLM and GCLC mRNAs remains almost unchanged while the abundance of other mRNAs reduces to varying degrees. Overall, the changes of gene expression in third trimester in comparison to the first one estimated by RT-qPCR and microarray coincide while the former data are more informative for the limited group of genes. The data provide the basis for further research of these genes expression and phenotype of human placenta in health and disease.

Influence of oxidative stress on the level of genes expression TGFB1 and HGF in rat liver upon long-term gastric hypochlorhydria and administration of multiprobiotic Symbiter

K. O. Dvorshchenko, O. O. Bernyk, A. S. Dranitsina, S. A. Senin, L. I. Ostapchenko

Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: k21037@gmail.com

Free-radical processes upon long-term omeprazole-induced gastric hypochlorhydria in the rat liver were researched. Intensification of oxidative processes in the liver tissue upon gastric hypoacid state was established: overproduction of superoxide anion, hydrogen peroxide, the quantitative changes of lipid functional groups, increased level of lipid peroxidation products, and augmentation of xanthine oxidase activity. The expression of Tgfb1 gene increased, while the expression of Hgf gene was not detected upon long-term suppression of gastric acid secretion of hydrochloric acid by omeprazole that indicated possible development of liver fibrosis. Abovementioned parameters were only partially restored to control values in the case of simultaneous administration of multiprobiotic “Symbiter® acidophilic” concentrated with omeprazole, thus indicating the ability of this preparation to counteract the development of oxidative damages in liver tissues upon long-term gastric hypoacidic conditions.

Stress-responsive systems in rat pancreas upon long-term gastric hypochlorhydria and administration of multiprobiotic “Symbiter®”

K. O. Dvorshchenko, S. Ye. Vakal, A. S. Dranitsina, S. A. Senin, L. I. Ostapchenko

Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: k21037@gmail.com

The intensity of free-radical processes upon long-term omeprazole-induced hypoacidity in the rat pancreas was investigated. Significant violation of oxidative-antioxidative balance in pancreatic tissue upon gastric hypochlorhydria was established: overproduction of superoxide anion, quantitative changes of lipid functional groups, increased level of lipid peroxidation products, augmentation of xanthine oxidase, superoxide dismutase and glutathione transferase activity, as well as depletion of catalase, glutathione peroxidase activity and reduced glutathione content. The inflected expression of Cckbr gene in the rat pancreas upon these conditions was also observed, thus suggesting an increased risk of pathological changes development in the gland. Abovementioned parameters were only partially restored to control values in the case of simultaneous administration of multiprobiotic “Symbiter®” with omeprazole, thus indicating the ability of this preparation to efficiently counteract the development of oxidative damages in pancrea­tic tissues upon long-term hypoacidic conditions.

Expression of phosphoribosyl pyrophosphate synthetase genes in U87 glioma cells with ERN1 knockdown: effect of hypoxia and endoplasmic reticulum stress

O. H. Minchenko, I. A. Garmash, O. V. Kovalevska,
D. O. Tsymbal, D. O. Minchenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: ominchenko@yahoo.com

Activation of pentose phosphate pathway is an important factor of enhanced cell proliferation and tumor growth. Phosphoribosyl pyrophosphate synthetase (PRPS) is a key enzyme of this pathway and plays a central role in the synthesis of purines and pyrimidines. Hypoxia as well as ERN1 (from endoplasmic reticulum to nuclei-1) mediated endoplasmic reticulum stress response-signalling pathway is linked to the proliferation because the blockade of ERN1 suppresses tumor growth, including glioma. We studied the expression of different PRPS genes in glioma cells with ERN1 knockdown under hypoxic condition. It was shown that hypoxia decreases the expression of PRPS1 and PRPS2 genes in both types of glioma cells, being more pronounced in cells without ERN1 function, but PRPSAP1 and PRPSAP2 gene expressions are suppressed by hypoxia only in glioma cells with blockade of ERN1. Moreover, the blockade of endoribonuclease activity of ERN1 does not affect the expression of PRPS1 and PRPS2 as well as PPRS-associated protein genes in U87 glioma cells. At the same time, the induction of endoplasmic reticulum stress by tunicamycin in glioma cells with suppressed activity of ERN1 endoribonuclease decreases the expression level of PRPS1 and PRPS2 genes only. Results of this investigation clearly demonstrated that the expression of different genes encoding subunits of PRPS enzyme is affected by hypoxia in U87 glioma cells, but the effect of hypoxia is modified by suppression of endoplasmic reticulum stress signaling enzyme ERN1.

Comparative analysis of gene expression in normal and cancer human prostate cell lines

E. E. Rosenberg, G. V. Gerashchenko, V. I. Kashuba

State Key Laboratory of Molecular and Cellular Biology,
Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
e-mail: y.e.rozenberg@imbg.org.ua

Prostate cancer is one of the main causes of mortality in men with malignant tumors. The urgent problem was a search for biomarkers of prostate cancer, which would allow distinguishing between aggressive metastatic and latent tumors. The aim of this work was to search for differentially expressed genes in normal epithelial cells PNT2 and prostate cancer cell lines LNCaP, DU145 and PC3, produced from tumors with different aggressiveness and metas­tatic ability. Such genes might be used to create a panel of prognostic markers for aggressiveness and metastasis. Relative gene expression of 65 cancer-related genes was determined by the quantitative polymerase chain reaction (Q-PCR). Expression of 29 genes was changed in LNCaP cells, 20 genes in DU145 and 16 genes in PC3 cell lines, compared with normal line PNT2. The obtained data make it possible to conclude that the epithelial-mesenchymal cell transition took place, which involved the loss of epithelial markers, reduced cell adhesion and increased migration. We have also found few differentially expressed genes among 3 prostate cancer cell lines. We have found that genes, involved in cell adhesion (CDH1), invasiveness and metastasis (IL8, CXCL2) and cell cycle control (P16, CCNE1) underwent most changes. These genes might be used for diagnosis and prognosis of invasive metastatic prostate tumors.