Biopolym. Cell. 2023; 39(3):220-230.
Molecular Biomedicine
Effect of complexes of natural oligoribonucleotides with D-mannitol on the viability of cell cultures of different origin
1Kraievska I. M., 1Tkachuk Z. Yu.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143

Abstract

Aim. Investigation of the effect of acidic and salt forms of natural yeast RNA oligoribonucleotides and their complexes with D-mannitol (ORN-D-M) on the viability of cell cultures of different origins. Methods. MTT assay, differential trypan blue staining. Results. A dose-dependent inhibition of the viability of mouse melanoma B16 cells and human glioblastoma U251 cells under the influence of ORN-D-M was established, as well as a dependence on the treatment time. U251 cells were more sensitive to the effect of the drug: at 48-hour treatment, the IC50 for this cell line was 1.0±0.3 mg/ml, whereas for B16 it was 2.5±0.4 mg/ml. At the same time, the inhibition of non-malignant canine kidney MDCK cells was observed only at high concentrations of the drug (IC50 – 6.1±1.1 mg/ml). The acidic form of ORN-D-M inhibited the viability of B16 and U251 cell cultures whereas the acidic form of ORN, salt forms of Na-ORN and Na-ORN-D-M showed a significantly lower inhibition activity. Conclusions. The acidic form of ORN-D-M has significant inhibitory activity against the malignant cells of mouse melanoma B16 and human glioblastoma U251. The acidic form of ORN-D-M is a promising drug for further antitumor studies in vitro and in vivo.
Keywords: natural oligoribonucleotides, ORN-D-M, anticancer activity, MTT assay

References

[1] Guallar-Garrido S, Julián E. Bacillus Calmette-Guérin (BCG) Therapy for Bladder Cancer: An Update. Immunotargets Ther. 2020; 9:1-11.
[2] Babjuk M, Böhle A, Burger M, Capoun O, Cohen D, Compérat EM, Hernández V, Kaasinen E, Palou J, Rouprêt M, van Rhijn BWG, Shariat SF, Soukup V, Sylvester RJ, Zigeuner R. EAU Guidelines on Non-Muscle-invasive Urothelial Carcinoma of the Bladder: Update 2016. Eur Urol. 2017; 71(3):447-61.
[3] Kremenovic M, Schenk M, Lee DJ. Clinical and molecular insights into BCG immunotherapy for melanoma. J Intern Med. 2020; 288(6):625-40.
[4] Tokunaga T, Yamamoto H, Shimada S, Abe H, Fukuda T, Fujisawa Y, Furutani Y, Yano O, Kataoka T, Sudo T. Antitumor activity of deoxyribonucleic acid fraction from Mycobacterium bovis BCG. I. Isolation, physicochemical characterization, and antitumor activity. J Natl Cancer Inst. 1984; 72(4):955-62.
[5] Yamamoto S, Yamamoto T, Kataoka T, Kuramoto E, Yano O, Tokunaga T. Unique palindromic sequences in synthetic oligonucleotides are required to induce IFN [correction of INF] and augment IFN-mediated [correction of INF] natural killer activity. J Immunol. 1992; 148(12):4072-6.
[6] Wang H, Rayburn E, Zhang R. Synthetic oligodeoxynucleotides containing deoxycytidyl-deoxyguanosine dinucleotides (CpG ODNs) and modified analogs as novel anticancer therapeutics. Curr Pharm Des. 2005; 11(22):2889-907.
[7] Komura F, Okuzumi K, Takahashi Y, Takakura Y, Nishikawa M. Development of RNA/DNA Hydrogel Targeting Toll-Like Receptor 7/8 for Sustained RNA Release and Potent Immune Activation. Molecules. 2020; 25(3):728.
[8] Pavlova IN, Konopatskova OM, Ekaterynushkin DA. Immunocorrection in patients with melanoma. Rus Biotherap J. 2016; 15(1):82-3.
[9] Tkachuk Z. Multiantivirus Compound, Composition and Method for Treatment of Virus Diseases. U.S. Patent 20,120,232,129, 16 April 2013.
[10] Melnichuk N, Kashuba V, Rybalko S, Tkachuk Z. Complexes of Oligoribonucleotides with d-Mannitol Modulate the Innate Immune Response to Influenza A Virus H1N1 (A/FM/1/47) In Vivo. Pharmaceuticals (Basel). 2018; 11(3):73.
[11] Marchyshak T, Yakovenko T, Shmarakov I, Tkachuk Z. The Potential Protective Effect of Oligoribonucleotides-d-Mannitol Complexes against Thioacetamide-Induced Hepatotoxicity in Mice. Pharmaceuticals (Basel). 2018; 11(3):77.
[12] Landskron G, De la Fuente M, Thuwajit P, Thuwajit C, Hermoso MA. Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res. 2014; 2014:149185.
[13] Rayburn ER, Ezell SJ, Zhang R. Anti-Inflammatory Agents for Cancer Therapy. Mol Cell Pharmacol. 2009; 1(1):29-43.
[14] Jana NR. NSAIDs and apoptosis. Cell Mol Life Sci. 2008; 65(9):1295-301.
[15] Tkachuk ZY, Levchenko SM. Mass spectrometric analysis of oligoribonucleotides of total yeast RNA. Dopov Nac akad nauk Ukr. 2021; 2:115-22.
[16] Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2):55-63.
[17] Motulsky HJ. Equation: log(inhibitor) vs. normalized response. GraphPad Curve Fitting Guide. Accessed 11 March 2023. Retrieved from https://www.graphpad.com/guides/prism/8/curve-fitting/reg_dr_inhibit_normalized.htm
[18] Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, Minor L. Cell Viability Assays. 2013 May 1 [updated 2016 Jul 1]. In: Markossian S, Grossman A, Brimacombe K, Arkin M, Auld D, Austin C, Baell J, Chung TDY, Coussens NP, Dahlin JL, Devanarayan V, Foley TL, Glicksman M, Gorshkov K, Haas JV, Hall MD, Hoare S, Inglese J, Iversen PW, Kales SC, Lal-Nag M, Li Z, McGee J, McManus O, Riss T, Saradjian P, Sittampalam GS, Tarselli M, Trask OJ Jr, Wang Y, Weidner JR, Wildey MJ, Wilson K, Xia M, Xu X, editors. Assay Guidance Manual . Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-.
[19] Zhernov YV. Analysis of cytotoxicity of humic substances of peloids. Bull Samara Sci Cen RAS. 2011; 13(1-8):1996-8.
[20] Stepanenko AA, Dmitrenko VV. Pitfalls of the MTT assay: Direct and off-target effects of inhibitors can result in over/underestimation of cell viability. Gene. 2015; 574(2):193-203.
[21] Shoemaker M, Cohen I, Campbell M. Reduction of MTT by aqueous herbal extracts in the absence of cells. J Ethnopharmacol. 2004; 93(2-3):381-4.
[22] Shalimov SA, Keĭsevich LV, Volchenskova II, Trokhimenko EP, Datsenko IuS, Evtushenko GV. Use of polyplatillen in the treatment of stage IV malignant neoplasms. Klin Khir (1962). 1992; 1:40-1.
[23] Tkachuk ZYu, Rybalko SL, Zharkova LD, Starosila DB, Semernikova LI. Anti-influenza effect of Nuclex. Rep NASU. 2010; 9:191-6.
[24] Tkachuk ZYu, Rybalko SL, Dyadyun DB, Starosila DB. Antiherpetic activity of Nuclex. Rep NASU. 2011; 4:162-8.
[25] Fedyanina LN, Besednova NN, Aminin DL, Epstein LM, Kalenyk TK. Experimental study of the antitumor activity of DNA from salmon milt and some of its mechanisms. Far Eastern Med J. 2006; 3:59-62.
[26] Dick RS, Hubbell HR. Sensitivities of human glioma cell lines to interferons and double-stranded RNAs individually and in synergistic combinations. J Neurooncol. 1987; 5(4):331-8.
[27] Taghavi Pourianazar N, Gunduz U. CpG oligodeoxynucleotide-loaded PAMAM dendrimer-coated magnetic nanoparticles promote apoptosis in breast cancer cells. Biomed Pharmacother. 2016; 78:81-91.
[28] Xun Y, Yang H, Kaminska B, You H. Toll-like receptors and toll-like receptor-targeted immunotherapy against glioma. J Hematol Oncol. 2021; 14(1):176.