Tag Archives: gene expression
Inhibitory action of methylene bisphosphonic acid on metabolic activity and viability of J774A.1 cells
D. O. Labudzynskyi*, E. P. Pasichna, O. I. Krynina, М. M. Veliky
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: labudzinskidmytro@gmail.com
Received: 01 February 2024; Revised: 22 March 2024;
Accepted: 24 March 2024; Available on-line: 30 April 2024
Bisphosphonates (BPs) are primary agents in the current pharmacological arsenal against osteoclast-related bone loss due to osteoporosis, Paget’s disease and bone tumors. Due to the lack of complete understanding of the molecular mechanism of their action in bone tissue and the overlap of key properties between BPs of different generations, integral studies of BPs inhibitory and antiresorptive properties are relevant today. The present work was carried out to establish a comprehensive study of the inhibitory effects of methylene bisphosphonic acid (MBPA) on the mevalonate pathway, metabolic activity and cell death in vitro compared to zoledronic acid (Zol). Farnesyl pyrophosphate synthase activity of MBPA-treated J774A.1 cells was inhibited by 80%, compared with a 79% reduction in Zol-treated samples. The ability of MBPA to decrease the percentage of viable cells in culture is slightly lower compared with Zol. After 24 h of incubation with lowest concentration, the percentage of inhibition of metabolic activity was 10.6 and 25%, respectively. After 48 h, these values were 34.8 and 55.6%, respectively. The inhibitory effects of MBPA and Zol on the intensity of incorporation of radioactively labeled precursor [14C]-acetate to the cholesterol fraction were 76.2 and 59.1%, respectively. In the case of isoprenoid fraction, the inhibitory effects were 40.9% and 51.2%, respectively. MBPA and Zol differently induced apoptosis in the J774A.1 cells culture, increased count of apoptotic cells in 2.4 and 6.3 times, and also increased the number of propidium iodide-positive cells in 7.4 and 19 times, respectively. MBPA and Zol also increased the number of TUNEL-positive cells in macrophage culture in 2.6 and 5 times, respectively. Zoledronate significantly reduced carbonic anhydrase 2 and nuclear factor of activated T-cells 1 gene expression levels compared to the MBPA action. Thus, the use of MBPA in future research and therapy of both cancer and osteoporosis looks promising due to lower cytotoxicity, high efficiency of mevalonate pathway inhibition and the possibility of dosage variation.
The role of microRNA-613 and its related genes in ovarian cancer
M. M. Mohammed, M. M. Ramzy*, S. S. Gaber,
H. A. Mohamed, M. R. Mohamed, A. M. Abdalla
Department of Biochemistry, Faculty of Medicine, Minia University, Egypt;
*e-mail: maggiemaher24@gmail.com
Received: 19 October 2022; Revised: 25 November 2022;
Accepted: 17 February 2023; Available on-line: 27 February 2023
Ovarian cancer (OC) is the most lethal gynecological cancer. Multiple genetic and epigenetic abnormalities have been detected in ovarian cancers. As microRNAs (miRNAs) play important roles in carcinogenesis, numerous researchers aim to determine the molecular mechanism that regulates the cancer cells proliferation and metastasis. In the current study, the expression of microRNA-613 and related KRAS and Ezrin genes was assessed by Real-time PCR in ovarian cancer tissue and the adjacent apparently normal tissues. Our results revealed that the expression of miRNA-613 was downregulated in ovarian cancer while the expression of KRAS and Ezrin was higher in cancer tissues compared to apparently normal ovarian tissues. There was a negative correlation between the expression of miRNA-613 and both KRAS and Ezrin genes expression and a positive correlation between KRAS and Ezrin gene expression. The results obtained confirm that miRNA-613 acts as a tumor-suppressive gene in ovarian cancer and can realize such impact through the expression of KRAS and Ezrin genes. These data contribute to the identification of potential biomarkers and novel targets for OC early detection and treatment.
TLR4 gene expression in patients with chronic suppurative otitis media
J. T. Venkataravanappa1, K. C. Prasad2, S. Balakrishna1*
1Department of Cell Biology and Molecular Genetics, Sri Devraj Urs Academy of Higher Education and Research, Karnataka, India;
2Department of Otorhinolaryngology, Sri Devaraj Urs Medical College, Sri Devraj Urs Academy of Higher Education and Research, Kolar-563103, Karnataka, India;
*e-mail: sharath@sduu.ac.in
Received: 23 July 2021; Accepted: 12 November 2021
Chronic suppurative otitis media (CSOM) is an infectious disease of the middle ear that involves inflammation and accumulation of fluid behind the eardrum. The pathogenesis of CSOM involves reduced bacterial clearance due to impairment of Toll-Like Receptor (TLR) 4 pathway. TLR4 receptor serves as the molecular sensor for bacterial endotoxin (lipopolysaccharide) and activates inflammatory cell signaling for clearing the bacteria. Previous studies have shown that the expression of TLR4 gene is reduced in middle ear epithelia of CSOM patients. Whether the expression of TLR4 gene is reduced in leukocytes is not known. In our present study we aim to compare the expression of the TLR4 gene in the blood samples of CSOM patients and healthy controls. A case-control study was carried out by involving 16 participants in each group. The levels of the TLR4 gene expression were measured by using the qRT-PCR method. The median (interquartile range) ΔCt values of the TLR4 gene expression was 4.85 (2.61- 8.55) in the patient group and 2.29 (-1.63-4.85) in healthy controls. Expression of the TLR4 gene in the leukocytes of CSOM patient group was reduced by ~5.9 fold compared to the control group and the difference was found to be significant (P = 0.01).
Changes in gene expression of lactate carriers (MCT1 and CD147) in cardiac muscle of diabetic male rats: the effect of dichloroacetate and endurance training
H. Rezaeinasab1*, A. Habibi1, M. Nikbakht1, M. Rashno2,3, S. Shakerian1
1Department of Exercise Physiology, Faculty of Sport Sciences, Shahid Chamran University of Ahvaz, Ahvaz, Iran;
2Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran;
3Department of Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
e-mail: hamed.rezaei2020@gmail.com
Received: 23 March 2020; Accepted: 25 June 2020
Lactate accumulation can activate the pathways of mitochondrial biogenesis in the heart muscle. The purpose of this study was to investigate the effects of Pyruvate Dehydrogenase Kinase 4 (PDK4) inhibition and endurance training on the gene expression of lactate carriers (MCT1 and CD147) in the cardiac muscle of STZ-diabetic rats. In this experimental study, 64 male Wistar rats were selected and randomly divided into eight groups after induction of diabetes with streptozotocin (STZ). The endurance training protocol was performed on a treadmill for 6 weeks. Intraperitoneal injection of DCA of 50 mg/ kg body weight was used for the inhibition of PDK4 in the myocardium. Gene expression were measured using real-time PCR. The two-way ANOVA test was used to analyze the data. The results of the study showed that after endurance training, the expression of MCT1, PDK4, and CD147 genes increased significantly in line with each other (P < 0.05), and by inhibition of PDK4 in the heart muscle, the expression of MCT1 and CD147 genes in the endurance training group + diabetes + DCA and in the diabetes group + DCA decreased significantly (P < 0.05). According to the results of this study, it can be concluded that the repeated accumulation of lactate caused by exercise training in diabetic patients decrease through mitochondrial adaptation by DCA injection and subsequently oxidative stress can be reduced in cardiac tissue of diabetic patients and heart efficacy can be increased.
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.
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.
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.