Tag Archives: apoptosis

Discovery of cell apoptosis regulation genes: Sydney Brenner, John E. Sulston and H. Robert Horvitz (The Nobel Prize in Physiology or Medicine 2002)

M. V. Grigorieva*, V. M. Danilova, S. V. Komisarenko

Palladin Biochemistry Institute, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: mvgrigorieva@biochem.kiev.ua

Received: 24 Nobember 2020; Accepted: 23 April 2021

The Nobel Prize in Physiology or Medicine 2002 was awarded to Sydney Brenner, Howard Robert Horvitz and John Edward Sulston for their discoveries concerning “genetic regulation of organ development and programmed cell death”. The scientists­ studied cell division and differentiation in the nematode Caenorhabditis elegans from the fertilized egg to the adult organism. As a result of their studies, key genes regulating organ development and programmed cell death (apoptosis) were identified, and corresponding genes were shown to exist in higher species, including humans. These discoveries shed light on the pathogenesis of many diseases and were important for further medical research.

In silico identification and biochemical validation of plausible molecular targets of 4-thiazolidinone derivative Les-3833 as a potential anticancer agent

L. Kоbylinska1*, D. Khylyuk2, I. Subtelna2,
M. Kitsera3, R. Lesyk2

1Department of Biochemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
2Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
3Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, Ukraine;
*e-mail: Kobylinska_Lesya@meduniv.lviv.ua; lesya8@gmail.com

Received: 16 January 2021; Accepted: 23 April 2021

Synthetic 4-thiazolidinone derivatives have a broad range of pharmacologic activities. Thus, 4-thiazolidinones are being investigated to create new molecules and develop active pharmaceutical substances for anticancer treatment. In our previous study, we investigated the pyrazoline-thiazolidinone-isatin conjugates, and determined that Les-3833 was the most active compound and might act through inhibition of PARP-, MAPK-, JNK-, Bcl-2-, CDK1/cyclin B, and/or the caspase family. The aim of this research was to perform molecular docking studies to enable the construction of a pharmacophore model for the Les-3833 compound and investigate probable biological targets. Pharmacophore modeling software packages performed molecular docking studies of probable biological targets and enabled the construction of a pharmacophore model. Docking models of Les-3833 with 11 enzymes involved in apoptotic mechanisms were studied. Based on the pharmacophore modeling results for all 11 enzymes, Les-3833 is predicted to be most active in Chk‑1, caspase-6, and caspase-8. Immunoblot analysis proved that the application of Les-3833 led to inhibition of Ser345 phosphorylation, which is induced by etoposide, the most important modification responsible for Chk‑1 activity. Taken together with the results of the docking studies, several mechanisms for the expression of antitumor activity by 4-thiazolidinones are suggested, and such multi-affinity is a characteristic feature of all these derivatives. The docking analysis confirmed the affinity of test compound Les-3833 for a topoisomerase II inhibitor and a high possibility of inhibitory interaction with Chk-1, caspase-6, and caspase-8.

Male and female rats differ in homeostatic shifts during pre-slaughter fear stress

S. S. Grabovskyi1, R. R. Panchuk2, N. R. Skorokhyd2, R. S. Stoika2*

1Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies, Lviv, Ukraine;
2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
*e-mail: stoika.rostyslav@gmail.com

Received: 19 June 2020; Accepted: 17 December 2020

In this investigation, the level of сortisol in blood plasma and  splenocytes apoptosis in male and female rats in response to fear stress created by animals’ waiting for different terms (20, 40, 60 min) before the slaughter were evaluated. A significant and dependent on the stress state duration increase in the level of cortisol in the blood plasma of  both male and female rats compared with the rats of control groups was found, which was more pronounced in females compared to males. The number of Annexin V-positive (apoptotic) cells was determined by FACS analysis. It was shown that the content of apoptotic splenocytes during the stress before slaughter was higher in females compared to males. The prolongation of  the stress period was accompanied by an increase in the content of apoptotic splenocytes in males and its decrease in females. The potential hormone-dependent attenuation of the mechanisms of adaptation to  stress before slaughter in female rats is discussed.

Effect of N-acetyl cysteine on oxidative stress and Bax and Bcl2 expression in the kidney tissue of rats exposed to lead

M. Gholami1, A. B. Harchegani2, S. Saeedian3,
M. Owrang4, M. R. Parvizi1*

1Department of Physiology, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran;
2Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
3Medical Genomic Research Center, Tehran Medicine Sciences Islamic Azad University, Tehran, Iran;
4Faculty of Medicine, Sari Branch, Islamic Azad University, Sari, Iran;
*e-mail: mparvizi@alumnus.tums.ac.ir

Received: 30 May 2020; Accepted: 17 December 2021

This study aimed to consider the lead-induced oxidative damage of the kidney of male rats and the role of antioxidant N-acetylcysteine (NAC) in preserving cells against Pb toxicity. Rats were randomly divided into five groups including G1 (control), G2 (single 70 mg/kg dose of Pb), G3 (continuous daily 2 mg/kg dosing of Pb for 4 weeks), G4 (single dose of Pb + 50 mg/kg NAC), and G5 (continuous daily dosing of Pb + 50 mg/kg NAC). The level of malonic dialdehyde (MDA) and total antioxidant capacity were measured spectrophotometrically.The level of Pb in  serum and kidney tissue was measured by atomic absorption spectroscopy. Expression of Bax and Bcl2 genes was estimated using RT-PCR.  It was shown that single and continuous exposure to Pb caused a considerable increase of Pb content in serum and kidney tissue of rats in G2 and G3 groups compared to other groups. NAC treatment significantly improved TAC values and decreased MDA values in the serum of rats exposed to Pb. Single and continuous Pb dosing caused a 3.9- and 13.1-fold increase in Bax expression and 1.5-fold and 2.1-fold decrease in Bcl2 expression in a kidney tissue respectively. The current study revealed that single and  especially continuous Pb exposure  was strongly associated with Pb accumulation, antioxidant depletion, oxidative stress and kidney cells apoptosis. NAC can help protect kidney tissue against Pb by elevating antioxidant capacity, mitigating oxidative stress and normalizing Bax and Bcl2 genes expression.

Vitamin D(3) regulates hepatic VEGF-A and apelin expression in experimental type 1 diabetes

D. O. Labudzynskyi1*, I. O. Shymanskyi1, O. O. Lisakovska1,
A. O. Mazanova1, L. V. Natrus2, M. M. Veliky1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Bogomolets National Medical University, Kyiv, Ukraine;
*e-mail: konsument3@gmail.com

Received: 09 July 2019; Accepted: 15 May 2020

The deficiency of vitamin D is associated with the risk of various chronic diseases, including diabetes mellitus and its complications. Given the strong genomic action of vitamin D hormone-active form, its deficiency can lead to dysfunction of cytokine signaling pathways, including those dependent on vascular endothelial growth factors (VEGFs) and apelin. The present study was carried out to define the link between VEGF-A and apelin expression in liver, hepatocytes viability and vitamin D status at experimental type 1 diabetes in mice. We established that chronic hyperglycemia at streptozotocin-induced diabetes was accompanied by a 2.2-fold decrease in 25OHD content in the serum and increased hepatocytes apoptosis and necrosis. Vitamin D deficiency correlated with increased apelin and VEGF-A (8- and 1.6-fold respectively) expression. Almost complete restoration of circulatory 25OHD content in serum was achieved at vitamin D3 treatment (800 IU/kg, per os, for 2 months) followed by reduced apelin and VEGF-A expression in liver and the decline of hepatocytes apoptosis. We conclude that vitamin D3 can be involved in cell survival, angiogenesis and fibrogenesis by modulating  VEGF-A and apelin dependent regulatory systems in diabetic liver.

Cytotoxic activity of the cluster rhenium compound with β-alanine ligands

K. V. Polokhina1, D. E. Kytova1, A. V. Shtemenko1, N. I. Shtemenko1,2

1Ukrainian State University of Chemical Technology, Dnipro, Ukraine;
2Dnipro University of Technology, Ukraine;
e-mail: n.shtemenko@i.ua

Received: 25 February 2019; Accepted: 29 November 2019

Earlier we have shown that cluster rhenium compounds not only inhibited tumor growth in vivo but also supported the antioxidant state of experimental animals. Further investigation of new dirhenium(III) and cluster rhenium compounds in human leukemic cells is of great importance. The aim of the recent work was to investigate the cytotoxic activity of the new cluster rhenium compound with β-alanine ligands [Re2Cl6(C3H7NO2)2]·1.5H2O (I) in the solutions and nanoliposomes alone and together with cisplatin in Jurkat cells. It was shown that I in solution had cytotoxicity close to cisplatin (LC50 = 2.06·10-6 M). The administration of the rhenium-platinum system with І showed  increased cytotoxic activity, especially high when both components of the system were in the mixed liposomes together (LC50 = 4.93·10-10 M). The new dirhenium dicarboxylate complex with zwitterionic amino acid ligands possesses an appreciable cytotoxic and proapoptotic activity against leukemic cells, especially in combination with cisplatin, guiding the search for novel active rhenium compounds and development of improved regimens for combined chemotherapy based on combination of rhenium-platinum compounds.

Apoptosis induction in human leukemia cells by novel 2-amino-5-benzylthiazole derivatives

N. S. Finiuk1,2, I. I. Ivasechko1, O. Yu. Klyuchivska1,
Yu. V. Ostapiuk3, V. P. Hreniukh2, Ya. R. Shalai2,
V. S. Matiychuk3, M. D. Obushak3,
A. M. Babsky2, R. S. Stoika1

1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Ivan Franko National University of Lviv, Biology Faculty, Lviv, Ukraine;
3Ivan Franko National University of Lviv, Chemistry Faculty, Lviv, Ukraine;
e-mail: stoika@cellbiol.lviv.ua

Received: 21 December 2018; Accepted: 20 March 2019

Derivatives of 2-amino-5-benzylthiazole are heterocyclic pharmacophores that exhibit different pharmacological activities including anticancer action. The mechanisms of such action of these compounds are not clear. The aim of the present study was to investigate apoptosis induction, particularly DNA damage in human leukemia cells, by the novel synthesized thiazole derivatives ‒ 2,8-dimethyl-7-(3-trifluoromethyl-benzyl)pyrazolo[4,3-e]thiazolo[3,2-a]pyrimidin-4(2H)-one (compound 1) and 7-benzyl-8-methyl-2-propylpyrazolo[4,3-e]thiazolo[3,2-a]pyrimidin-4(2H)-one (compound 2). Western-blot analysis, DNA comet assay in alkaline conditions, diphenylamine DNA fragmentation assay, agarose gel retardation, and methyl green DNA intercalation assays were used to study the effects of the studied compounds in human leukemia cells. These compounds induced PARP1 and caspase 3 cleavage in the leukemia cells, also increased the level of pro-apoptotic Bim protein and the mitochondrion-specific EndoG nuclease, and decreased the level of the anti-apoptotic Bcl-2 protein. They caused DNA single-strand breaks and DNA fragmentation in the leukemia cells without direct DNA binding or DNA intercalation. Thus, novel 2-amino-5-benzylthiazole derivatives may be promising agents for apoptosis induction in the targeted human leukemia cells.

Disturbance of the transmembrane phosphatidylserine asymmetry in hepatocytes as an apoptosis marker under the action of xenobiotics on rats

O. A. Nakonechna, L. A. Babijchuk, A. I. Bezrodna

Kharkiv National Medical University, Kharkiv, Ukraine;
e-mail: bezrodnaya.ai@gmail.com

It has been reported that unfavorable chemical environmental factors affect the functional state of liver, activate free radical processes against the background of the reduced antioxidant activity, change physico-chemical properties and membrane phospholipid composition of  hepatocytes. The aim of our research was to estimate phosphatidylserine distribution in the phospholipid bilayer of hepatocyte membranes and apoptosis stages in hepatocytes of rats under the influence of surfactants: ethyleneglycol (EG), polyethyleneglycol 400, (PEG-400) and polypropyleneglycol (PPG) at a dose of 1/10 DL50. It was found in the  subacute toxicological experiment on rats that the investigated xenobiotics EG, PEG-400 and PPG at a dose of 1/10 DL50 caused phosphatidylserine translocation to the outer membrane in the phospholipid bilayer of hepatocytes. This is a specific signal for macrophages aiming at recognition and elimination of apoptotic cells. Analysis of cell death modes under the influence of the investigated xenobiotics at a dose of 1/10 DL50 revealed that the intake of xenobiotics was associated with  an increase in the amount of apoptotic/necrotic hepatocytes.

Biochemical aspects of the combined use of taxanes, irradiation and other antineoplastic agents for the treatment of anaplastic thyroid carcinoma

V. M. Pushkarev, O. I. Kovzun, V. V. Pushkarev, B. B. Guda, M. D. Tronko

SI V. P. Komisarenko Institute of Endocrinology and Metabolism, NAMS of Ukraine, Kyiv;
e-mail: pushkarev.vm@gmail.com

The review summarizes the results of the cycle of own research and literature data on biochemical mechanisms of combined action of taxanes with γ-irradiation and other antineoplastic agents on one of the most aggressive types of human cancer – anaplastic thyroid carcinoma. Antagonistic interplay between taxanes and irradiation at the level of apoptotic mechanisms and regulators of the cell cycle are discussed. The effectiveness and prospects of using low concentrations of taxanes and low doses of fractional γ-irradiation are substantiated. Attention is paid to the role of inflammation and its key factor – NF-κB in the genesis of thyroid carcinomas and their treatment. Directions for further research are outlined.

Biochemical mechanism of the o,p’-DDD effect on the adrenal cortex

A. S. Mikosha, O. I. Kovzun

V. P. Komisarenko Institute of Endocrinology and Metabolism, National Academy of Medical Sciences of Ukraine, Kyiv;
e-mail: asmikosha@gmail.com

o,p’-Dichlorodiphenyldichloroethane (o,p’-DDD, mitotane) is used in the treatment of adrenocortical cancer and Cushing’s disease. This medicine induces numerous biochemical changes in the adrenal cortex, as well as disorder in the mitochondrial structure. Therewith, the level of produced corticosteroid hormones is significantly reduced. One of the possible causes can be a decrease in the NADPH level due to inhibition of the activity of its reduction system and increased NADPH consumption during the glutathione reduction catalyzed by glutathione reductase. o,p’-DDD is partially metabolized in the adrenal glands, and   the main metabolite (in terms of quantity) is o,p’-dichlorodiphenylacetic acid. However, attempts to find a physiologically active component among metabolites were unsuccessful. The most pronounced changes caused by o,p’-DDD were found in the mitochondria of the adrenal cortex. The respiration at the level of IV and I complexes is suppressed, the protein content of these complexes decreases. The phospholipid composition of the tissue altered and the concentration of diphosphatidylglycerol, the most important component of mitochondrial membranes, decreased. In our opinion, o,p’-DDD, owing to its high lipophilicity, accumulates in the mitochondria membranes and causes conformational disorder followed by disorder in mitochondrial functions. It was shown that o,p’-DDD acts as an inhibitor of acyl-CoA-cholesterol acyltransferase (ACAT, SOAT1). Therefore, adenocorticocytes accumulate free cholesterol, causing endoplasmic reticulum stress, mitochondrial swelling and caspases activation. Increased apoptosis leads to a decline in adrenal function and to a decrease in weight of adrenal glands.