INDUCTION OF APOPTOSIS OF PROSTATE CANCER CELLS BY THE EXTRACT OF THE FRESHWATER SPONGE

DOI: https://doi.org/10.29296/24999490-2022-04-08

A.A. Nikolaev, N.I. Gudinskaya, M.V. Ushakova
FGBOU VO «Astrakhan State Medical University» of the Ministry of Health of the Russian Federation,
121 Bakinskaya str., Astrakhan, 414000, Russian Federation

Introduction. Prostate cancer (PC) is by far the most commonly diagnosed malignant neoplasm of the urinary system. The pathogenesis and molecular mechanisms of prostate cancer remain incompletely understood. There are reports of the presence in the extracts of several species of marine and freshwater sponges of bioactive metabolites that reduce the proliferative activity of cancer cells and, in particular, prostate cancer cells. Purpose. The purpose of our study was the effect of freshwater badyagi extract on the viability and apoptosis of prostate cancer cells LNCAP clone FSK (ECACC 89110211) and the associated mechanism. Material and methods. In the work, we used an extract of the Commonweed (EPHYDATIA FLUVIATILIS), LNCAP cells of the FSK clone (ECAC 89110211), Analysis of cell proliferation was performed by a standard MTT test, Analysis of apoptosis was performed by flow cytometry using an Attune® NxT flow cytometer ANNEXIN Apoptosis Detection Kit V-FITC APOPTOSIS DETECTION KIT I (BD PharmingemTM, USA) was used for further confirmation of apoptosis. To determine the effect of bodyaga extract on the signaling pathway involved in this study, pro-apoptotic and anti-apoptotic proteins in LNCAP cells were assessed by Western blot analysis. Results and discussion. A preliminary study of the effect of EBR on LNCAP cell proliferation showed that EBR exhibited the maximum growth inhibition effect on LNCAP cells starting from the second day of incubation. The further experiment pursued the goal of elucidating the dependence of the inhibitory effect on the concentration of EBR in the cultivation medium. A strong dose-response relationship between EBR and LNCAP cell viability was shown. However, it was shown that EBR caused a slight significant increase in cell viability between 0 µg/ml (control) and 6.25 µg/ml. This reaction can be explained by the well-known hormesis effect. An assessment of the safety profile of EBR showed a selectivity index of 7.24. It was shown that it was apoptosis that contributed to the decrease in the viability of LNCAP prostate cancer cells after exposure to EBR. Moreover, there is a direct dependence of the level of apoptosis on the concentration of EBR in the culture liquid. Western blot analysis of LNCAP cells treated with various concentrations (0, 50, 100 and 200 mg/ml) of EBR to assess the expression of Bcl-2, Bax proteins showed that the level of Bax was rising and Bcl-2 was falling. Conclusion. EBR has an inhibitory effect on the growth and viability of prostate cancer cells. causing apoptosis of prostate cancer cells. EBR treatment also increased the Bax/Bcl2 expression ratio, indicating involvement of a mitochondria-mediated intrinsic apoptotic pathway.
Keywords: 
prostate cancer, apoptosis induction, bodyaga extract

Список литературы: 
  1. Винник Ю.Ю., Андрейчиков А.В., Климов Н.Ю. Современное представление о диагностике рака простаты. Урология. 2017; 2: 110–52. [Vinnik Yu.Yu., Andrejchikov A.V., KlimovN.Yu. Sovremennoe predstavlenie o diagnostike raka prostaty`. Urologiya. 2017; 2: 110–52 (In Russian)].
  2. Ottinger S., Klöppel A., Rausch V., Liu L., Kallifatidis G., Gross W., Gebhard M.M., Brümmer F., Herr I. Targeting of pancreatic and prostate cancer stem cell characteristics by Crambe crambe marine sponge extract. Int. J. Cancer. 2012; 130 (7): 1671–81. https://doi.org/ 10.1002/ijc.26168.
  3. Николаев А.А., Выборнов С.В. Торможение роста клеток культуры рака простаты LNCAP аналогами полиаминов. Современные проблемы науки и образования. 2016; 6: 1–8. [Nikolaev A.A., Vy`bornov S.V. Tormozhenie rosta kletok kul`tury` raka prostaty` LNCAP analogami poliaminov. Sovremenny`e problemy` nauki i obrazovaniya. 2016; 6: 1–8. URL: http://science-education.ru/ru/article/view?id=25573 (In Russian)].
  4. Niks M., Otto M. Towards an optimized MTT assay. J. Immunol Meth 1990; 130 (1): 149–51 https://doi.org/10.1016/0022-1759(90)90309-j.
  5. Shi J.M., Huang H.J., Qiu S.X., Feng S.X., Li X.E. Griffipavixanthone from Garcinia oblongifolia champ induces cell apoptosis in human non-small-cell lung cancer H520 cells in vitro. Molecules. 2014; 19 (2): 1422–31. https://doi.org/ 10.3390/molecules19021422
  6. Pillai-Kastoori L., Schutz-Geschwender A.R., Harford J.A. A systematic approach to quantitative Western blot analysis. Anal Biochem. 2020; 593: 113608. https://doi.org/10.1016/j.ab.2020.113608.
  7. Jodynis-Liebert J., Kujawska M.. Biphasic Dose-Response Induced by Phytochemicals: Experimental Evidence. J. Clin. Med. 2020; 9 (3): 2–28. https://doi.org/10.3390/jcm9030718.
  8. Razumov A., Zav’yalov E.L., Troitskii S.Yu., Romashchenko A.V., Petrovskii D.V., Kuper K.E., Moshkin M.P. Selective Cytotoxicity of Manganese Nanoparticles against Human Glioblastoma Cells. Bulletin of Experimental Biology and Medicine. 2017; 163 (3): 561–5. https://doi.org/10.1007/s10517-017-3849-0