Влияние ДНК-тропных антиканцерогенных соединений на механизмы регуляции экспрессии генов

The presented review is devoted to the analysis of molecular mechanisms of action for different natural DNA-tropic compounds with established tumor preventive activity. Here we present their cancer preventive effects observed in vivo, mechanisms of DNA binding, influence on epigenetic regulation and “housekeeping” protein function. Additionally, the influence of these compounds on DNA helix parameters is discussed that should impact on epigenetic regulation of gene expression and formation of topologically associated domains.Обзор посвящен анализу молекулярных механизмов действия ряда природных ДНК-тропных соединений с установленной антиканцерогенной активностью. Приведены данные исследований антиканцерогенного действия этих соединений в экспериментах in vivo, рассмотрены механизмы их связывания с ДНК, влияния на метилирование ДНК и модификацию гистонов, способность к ингибированию функций ферментов «домашнего хозяйства». Кроме того, проанализированы возможные эффекты этих соединений на характеристики дуплекса ДНК, что должно иметь значение для эпигенетической регуляции экспрессии генов и формирования топологически ассоциированных доменов

Influence of natural DNA-binding compounds with cancer preventive activity on chromatin structure and interferon signaling

Blokhin NMRCO, Department of Chemical Carcinogenesis, Moscow, Russia1 Roswell Park Cancer Institute, Department of Cell Stress Biology, Buffalo, USA2 Ryazan State Medical University, Ryazan, Russia3 RUDN University, Moscow, Russia4 Chromatin-destabilizing effects of the natural DNA-binding small molecules Curcumin, Quercetin, Resveratrol, Genistein, Fisetin, Coumarin, Apigenin, EGCG, Thymoquinon and Naringenin on chromatin structure were analyzed by histone H1 and chaperone of histone SSRPtranslocation in cell nuclear. Using live cell fluorescent imaging we demonstrated that all the natural DNA-binding compounds studied were able to cause the appearance of cell fractions with high histone H1 amount localized in nucleolus. Visible changes in location of SSRP in cells treated with all the phytonutrients was also observed by live cell fluorescent imaging and for the number of compounds its translocation from nucleoles was confirmed by Western Blotting. It should be noted that dynamics of the phytonutrients influence on H1 and SSRP translocation varied greatly. We revealed activation of INF signaling by Genistein, Curcumin, Kaempferol, Sanguinarin, Naringenin, Resveratrol, Fisetin and Quercetin using flow cytometry of the HeLa-TI cells with ISRE-mCherry reporter

Activation of interferon-α signaling by resveratrol, genistein and quercetin

Resveratrol, genistein and quercetin from the group of polyphenols from secondary plant metabolites reveal cancer preventive and antivirus effects realized via their pleiotropic influence on the different macromolecules in cells. These compounds can interact with DNA without the formation of covalent bonds. This process is usually followed by changes in spatial, physical-chemical and structural DNA characteristics that can result in disfunction of DNA metabolism enzymes and chromatin destabilization. Similar effects were described for anticancer drug Curaxine CBL0137 in association with activation of interferon-α signaling. We demonstrated dose-dependent stimulating effects of resveratrol, genistein and quercetin on interferon-α signaling using HeLa cells expressed mCherry protein with interferon-stimulated response elements (ISRE) in promoter. Furthermore, it was shown by live-cell fluorescent microscopy in HT1080 cells with mCherry-labeled histone H1.5 that described polyphenols induced the redistribution of this linker histone in cell nuclei. The data obtained suggest an existence of DNA-dependent mechanism of anticancer effects of plant polyphenols and a need for further study of crosslinks between the polyphenols’ influence on chromatin structure and the changes in genome function, in particular, induction of interferon-interferon-α signaling. © 2019, Tomsk National Research Medical Center of the Russian Academy of Sciences. All rights reserved

Buccal Micronucleus Cytome Assay for the Evaluation of Cytogenetic Status of Healthcare Professionals Contacting with Anti-Cancer Drugs

Abstract: Healthcare professionals of chemotherapy departments are in a regular contact with anticancer drugs that have genotoxic properties. The analysis of the cytogenetic status of healthcare professionals (54 individuals) was for the first time carried out using a new promising noninvasive method, buccal micronucleus cytome assay with the calculation of integral indices. The effect was estimated with different duration of exposure to chemotherapy drugs (after an employee’s vacation, one month after vacation, and before vacation). Statistically significant increases in almost all studied indices of cytogenetic damages, proliferation, and apoptosis as compared with the control group of office staff (47 individuals) were detected. Among healthcare professionals, an increase in the portion of individuals with an increased level of cytogenetic abnormalities with the maximum exposure duration from 24 to 38% (11% in the control) was established. Among cancer patients (9 individuals), the portion of individuals with a high level of cytogenetic abnormalities was 56% before the next course of chemotherapy and 87% after it. © 2022, Pleiades Publishing, Inc

Molecular mechanisms of anticancer activity of n-glycosides of indolocarbazoles lcs-1208 and lcs-1269

Novel indolocarbazole derivatives named LCS were synthesized by our research group. Two of them were selected as the most active anticancer agents in vivo. We studied the mechanisms of anticancer activity in accordance with the previously described effects of indolocarbazoles. Cytotoxicity was estimated by MTT assay. We analyzed LCS-DNA interactions by circular dichroism in cholesteric liquid crystals and fluorescent indicator displacement assay. The effect on the activity of topoisomerases I and II was studied by DNA relaxation assay. Expression of interferon signaling target genes was estimated by RT-PCR. Chromatin remodeling was analyzed–the effect on histone H1 localization and reactivation of epigenetically silenced genes. LCS-induced change in the expression of a wide gene set was counted by means of PCR array. Our study revealed the cytotoxic activity of the compounds against 11 cancer cell lines and it was higher than in immortalized cells. Both compounds bind DNA; binding constants were estimated—LCS-1208 demonstrated higher affinity than LCS-1269; it was shown that LCS-1208 intercalates into DNA that is typical for rebeccamycin derivatives. LCS-1208 also inhibits topoisomerases I and IIα. Being a strong intercalator and topoisomerase inhibitor, LCS-1208 upregulates the expression of interferon-induced genes. In view of LCSs binding to DNA we analyzed their influence on chromatin stability and revealed that LCS-1269 displaces histone H1. Our analysis of chromatin remodeling also included a wide set of epigenetic experiments in which LCS-1269 demonstrated complex epigenetic activity. Finally, we revealed that the antitumor effect of the compounds is based not only on binding to DNA and chromatin remodeling but also on alternative mechanisms. Both compounds induce expression changes in genes involved in neoplastic transformation and target genes of the signaling pathways in cancer cells. Despite of being structurally similar, each compound has unique biological activities. The effects of LCS-1208 are associated with intercalation. The mechanisms of LCS-1269 include influence on higher levels such as chromatin remodeling and epigenetic effects. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Palivizumab: Four seasons in Russia

In 2010, the Russian Federation (RF) registered palivizumab - innovative drug, based on monoclonal antibodies for passive immunization of seasonal respiratory syncytial virus (RSV) infection in children of disease severe progress risk group, which include primarily premature infants, children with bronchopulmonary dysplasia and hemodynamically significant congenital heart disease. Currently, palivizumab is included in the list of recommended medicines and medical care standards of different countries, including Russia. In the review the results of Russian research on the progress of RSV infection, its epidemiology and immunization experience gained over the 2010-2014 period are summarized in relation to the foreign data. During the four epidemic seasons palivizumab immunization covered more than 3,200 children of severe RSV infection risk group with a progressive annual increase in the number of patients who received the drug. Geography of palivizumab immunization is also greatly expanded in our country during this time. If during the first two seasons measures of immunization were taken mainly in Moscow and St. Petersburg, at the present time, thirty one territorial entities of the Russian Federation have the experience in the drug application. Analysis of the results of RSV infection immunization (made in several regions) confirms the high clinical efficacy and palivizumab safety already demonstrated in international studies. In addition, the analysis presents the potential to improve the efficiency of the integrated RSV infection immunization programs, realizing in the establishment of high-risk child group register, adequate counseling for parents, as well as the development of the routing of patients and coordination of interaction between different health institutions during the immunization. © 2014, Izdatel'stvo Meditsina. All rights reserved