PATHOMORPHOSIS AND MECHANISMS OF DEATH OF TUMOR CELLS IN CELL CULTURES AND INOCULATED TUMORS UNDER THE INFLUENCE OF DRUG HEDGE HYSSOP EXTRACT

• Published in: Cardiometry no. 24; pp. 41 - 42
• Main Authors: Navolokin, N A, Mudrak, D A, Bucharskaya, A B, Polukonova, N V, Maslyakova, G N
• Format: Journal Article
• Language: English
• Published: Moscow Russian New University 30.11.2022
• Subjects: Apoptosis; Autophagy; Cancer therapies; Cell culture; Cell cycle; Cervical cancer; Drugs; Flavonoids; Kidney cancer; Ligands; Morphology; Prostate cancer; Sarcoma; Tumors
• ISSN: 2304-7232
• DOI: 10.18137/cardiometry.2022.24.conf.22

At present, the incidence of cancer tends to dramatically increase, and the existing advanced methods of cancer treatment are not always effective [1]. In addition, the disadvantages of the antitumor drugs used are their toxic effect made on the intact organs and tissues of the body, as well as the development of tumor resistance to them [2]. All this makes it necessary to search for new, safer and more effective drugs. In this case, plant-derived medicines have minimal side effects, so they are given special attention today. Plant-based drugs can be used both for the treatment of tumors as monotherapy and also in combination treatment to protect normal cells (including bone marrow stem cells) in a standard course of chemotherapy and radiation therapy [1,2, 3]. According to some researchers, the most promising group of this sort of medical drugs are flavonoids, since they produce the greatest number of biological effects, which favorably influence outcomes of treatment of neoplasms [2,3]. The aim of our research was to reveal some features of the pathomorphosis of tumors of different histogenesis, identify possible mechanisms of the tumor cell death and the resistance of tumor cells in experiments in cell cultures of human tumors (in vitro) and inoculated tumors in animals (in vivo) under the influence made by the flavonoid-containing hedge hyssop extract (Gratiola officinalis). Materials and methods. The methodology of our research work based on a combination of experimental and analytical methods, as well as on a systematic and complex analysis of the already available data and obtained fresh evidence data. The methods and techniques used in our work were as follows: the cultural method, fluorescent staining techniques, flow cytometry methods, electron microscopy, morphological methods, morphometric methods, histochemical (Pas-reaction, WGA, MHS staining, DNA and RNA staining according to Brache) and immunohistochemical methods (markers of Ki67, EGFR, VEGF, p53, Bax, Bcl-2, CD95 (Fas/APO-1), Fas-ligand and LC3B; statistical processing of results. In order to study the death of tumor cells, different groups of bio-flavonoids were used: flavonones (extract of Helichrysum arenarium), flavonols (extract of Gratiola officinalis), anthocyanins (extracts of the anthocyanin form of Zea mays). The objects of our study were cell cultures as given below: SPEV - culture of epithelial cells of the kidney of the pig embryo; HeLa - cervical cancer; SK-BR-3, MCF-7 - breast adenocarcinoma; Jurkat - T-cell lymphoblastic leukemia; A549 - lung carcinoma; PC-3 - prostate carcinoma; HCT-116 - colorectal carcinoma; A498, Caki and Sn12c - kidney carcinomas; tissues of inoculated tumors (sarcoma 45, kidney cancer PA and liver cancer PC-1); cells of the internal organs and the brain, fixed in 10% buffered neutral formalin solution and embedded in paraffin. For the statistical data processing, the normality of the distribution of the indicators in the groups was tested using the Shapiro-Wilk test. To compare the parameters in the groups, the Cramer-Welch test (T) was applied. Results. Regardless of the histological structure of the inoculated tumor in the animals, with the introduction of hedge hyssop extract, grade 2-3 pathomorphosis of 2-3 developed in the tumor: the proliferative activity decreased (Ki67; EGFR), the cell cycle stopped and the cells entered the G0 phase (Ki67), angiogenesis (VEGF) was blocked, and apoptosis was activated (p53, Bax, CD95 (Fas/APO-1), Fas-ligand), and protective autophagy was blocked (LC3B). The features of the pathomorphosis in the inoculated sarcoma 45 involved the replacement of the tumor tissue by the fibrous tissue, and in case of kidney cancer PA the development of giant cell pathomorphosis. Conclusion. Thus, in our experiments in vitro and in vivo, as a result of a comprehensive morphological explorations of structural changes and alterations in the tumor cells under the action made by the hedge hyssop extract, we found that apoptosis caused the typical cell death, realized through the caspase-3 activation mechanism, and that the expression of the LC3B protein, an autophagy marker, was a morphological criterion and predictor of the tumor resistance to antitumor drugs.