Log in

Biotechnology of Cultivation of Rhaponticum Carthamoides (willd.) Suspension Cells: A Prospective Source of Antitumor Substances

  • CROP PRODUCTION, PLANT PROTECTION, AND BIOTECHNOLOGY
  • Published:
Russian Agricultural Sciences Aims and scope

Abstract

To preserve the species diversity of Rhaponticum carthamoides, a Red Book plant that is a promising source of antitumor metabolites, it is important to grow cell cultures in vitro. The aim of the work is to select the optimal nutrient medium for growing suspension cells of Rhaponticum carthamoides, varying the mineral composition of the nutrient medium and growth regulators. To obtain suspension cells in vitro, nutrient media of various mineral compositions (Murashige–Skoog, Gamborg, and Schenk–Hildebrandt) and growth regulators (auxin: 3-indoleacetic acid (IAA); cytokinin: kinetin) were used. Cultivation parameters were as follows: 26°C, 60–70% humidity, 16-h daylight period and 21-day subculture cycle. For the obtained cell cultures, the viability, growth index, and specific growth rate were assessed. The results showed that the highest values of the studied parameters were observed when growing suspension cells on a nutrient medium with a Schenck–Hildebrandt mineral composition (SH) (77–82% viability of suspension cells, 6.11‒7.14 growth index, 0.17–0.21 day–1 specific growth rate). Methanol extracts were prepared from selected and prelyophilized cell samples and analyzed by high performance liquid chromatography. The results showed that suspension cultures (the best indicators were observed in sample no. 12 grown on SH medium supplemented with IAA and kinetin), as well as the control sample (methanol extract of crushed plant root), contained sitosterol (0.26 ± 0.01 mg/g in control; 1.54 ± 0.02 mg/g in no. 12), isofraxidine (1.15 ± 0.02 mg/g in control; 3.10 ± 0.04 mg/g in no. 12), chlorogenic (44.54 ± 0.14 mg/g in control; 66.87 ± 0.02 mg/g in no. 12), and coffee (50.15 ± 0.03 mg/g in control; 54.73 ± 0.06 mg/g in no. 12) acid. The extracts of suspension cultures (100 mg/mL) exhibited antitumor properties: a cytotoxic effect on tumor cells (PANC-1, LBR2, MDAMB-231, NT-29). Cell cultures grown on SH medium supplemented with IAA and kinetin exhibited the most pronounced antitumor properties since they contained a greater amount of biologically active substances in their composition. When the extract of sample no. 12 was added, the decrease in the viability of tumor cells was achieved by 40.57–53.23%. In the course of the work, the composition of the nutrient medium was selected, which allows one to grow the biomass of suspension cultures of Rhaponticum carthamoides cells, which exhibits antitumor properties.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1.

Similar content being viewed by others

REFERENCES

  1. Skała, E., Synowiec, E., Kowalczyk, T., et al., Rhaponticum carthamoides transformed root extract has potent anticancer activity in human leukemia and lung adenocarcinoma cell lines, Oxid. Med. Cell. Longevity, 2018, vol. 2018, art. ID 8198652. https://www.hindawi.com/journals/omcl/2018/8198652/#copyright. Cited October 16, 2021. https://doi.org/10.1155/2018/8198652

  2. Kokoska, L. and Janovska, D., Chemistry and pharmacology of Rhaponticum carthamoides: A review, Phytochemistry, 2009, vol. 70, no. 7, pp. 842–855. https://doi.org/10.1016/j.phytochem.2009.04.008

  3. Solov’eva, M.A., Factors affecting the content of 20-hydroxyecdisone in Leuzea carthamoides (review), Vestn. Permsk. Fed. Issled. Tsentra, 2020, no. 4, pp. 62–75. https://doi.org/10.7242/2658-705X/2020.4.6

  4. Havlik, J., Budesinsky, M., Kloucek, P., et al., Norsesquiterpene hydrocarbon, chemical composition and antimicrobial activity of Rhaponticum carthamoides root essential oil, Phytochemistry, 2009, vol. 70, no. 3, pp. 414–418. https://doi.org/10.1016/j.phytochem.2008.12.018

    Article  CAS  PubMed  Google Scholar 

  5. Skała, E., Toma, M., Kowalczyk, T., et al., Rhaponticum carthamoides transformed root extract inhibits human glioma cells viability, induces double strand DNA damage, H2A.X phosphorylation, and PARP1 cleavage, Cytotechnology, 2018, vol. 70, no. 6, pp. 1585–1594. https://doi.org/10.1007/s10616-018-0251-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Skała, E., Makowczyńska, J., Wieczfinska, J., et al., Caffeoylquinic acids with potential biological activity from plant in vitro cultures as alternative sources of valuable natural products, Curr. Pharm. Des., 2020, vol. 26, no. 24, pp. 2817–2842. doi 2826666200212115826https://doi.org/10.2174/138161

  7. Andreev, B.G., Volobaev, P.A., and Gorbunova, I.A., et al., Krasnaya kniga Kuzbassa (Red Book of Kuzbass), Kemerovo: VEKTOR-PRINT, 2021, vol. 1.

  8. Reshetnikov, V., Spiridovich, E., Fomenko, T., et al., Plant biotechnology as a way of biosynthetic potential rational use, Nauka Innovatsii, 2014, vol. 5, no. 135, pp. 21–25.

    Google Scholar 

  9. Skała, E., Rijo, P., Garcia, C., et al., The essential oils of Rhaponticum carthamoides hairy roots and roots of soil-grown plants: chemical composition and antimicrobial, anti-inflammatory, and antioxidant activities, Oxid. Med. Cell. Longevity, 2016, vol. 2016, art. ID 8505384. https://www.hindawi. com/journals/omcl/2016/8505384/. Cited October 16, 2021. https://doi.org/10.1155/2016/8505384

  10. Skała, E., Kicel, A., Olszewska, M.A., et al., Establishment of hairy root cultures of Rhaponticum carthamoides (Willd.) Il** for the production of biomass and caffeic acid derivatives, BioMed Res. Int., 2015, vol. 2015, art. ID 181098. https://www.hindawi.com/journals/bmri/2015/181098/ Cited October 15, 2021. https://doi.org/10.1155/2015/181098

  11. Velichko, N.A. and Smol’nikova, Ya.V., The influence of technological parameters of extraction on the yield of extractable substances from leuzea safflower (Rhaponticum carthamoides), Vestn. Krasnoyarsk. Gos. Agrar. Univ., 2017, vol. 133, no. 10, pp. 73–78.

  12. Sib. Gos. Tekhnol. Univ., Patent № 2130709, 1999.

  13. Murashige, T. and Scoog, F., A Revised medium for rapid growth and bio assays with tobacco tissue culture, Physiol. Plant., 1962, vol. 15, no. 3, pp. 473–497.

    Article  CAS  Google Scholar 

  14. Gamborg, O.L., Miller, R.A., and Ojima, O., Nutrient requirements of suspension cultures of soybean root cells, Exp. Cell Res., 1968, vol. 50, no. 1, pp. 151–158.

    Article  CAS  Google Scholar 

  15. Asyakina, L.K., Dyshlyuk, L.S., Pozdnyakova, A.V., et al., Selection of a system for apparatus cultivation of suspension cultures of cells and root cultures in vitro in  bioreactors of various designs and volumes, Vestn. Yuzhno‑Ural. Gos. Univ., Ser.: Pishch. Tekhnol., 2021, vol. 9, no. 3, pp. 41–52. https://doi.org/10.14529/food210305

    Article  Google Scholar 

  16. Skała, E., Sitarek, P., Toma, M., et al., Inhibition of human glioma cell proliferation by altered Bax/Bcl-2-p53 expression and apoptosis induction by Rhaponticum carthamoides extracts from transformed and normal roots, J. Pharm. Pharmacol., 2016, vol. 68, no. 11, pp. 1454–1464. https://doi.org/10.1111/jphp.12619

    Article  CAS  PubMed  Google Scholar 

  17. Yang, I, Asyakina, L.K., Babich, O.O., et al., Physicochemical properties and biological activity of extracts of dried biomass of callus and suspension cells and in vitro root cultures, Tekh. Tekhnol. Pishch. Proizvod., 2020, vol. 50, no. 3, pp. 480–492. https://doi.org/10.21603/2074-9414-2020-3-480-492

    Article  Google Scholar 

  18. Kweon, B., Han, Y.H., Kee, J.Y., et al., Effect of Angelica gigas Nakai ethanol extract and decursin on human pancreatic cancer cells, Molecules (Switzerland), 2020, vol. 25, no. 9, art. ID 2028. https://www. mdpi.com/1420-3049/25/9/2028#cite. Cited October 16, 2021. https://doi.org/10.3390/molecules25092028

  19. Zaushintsena, A.V., Milentyeva, I.S., Babich, O.O., et al., Quantitative and qualitative profile of biologically active substances extracted from purple echinacea (Echinacea purpurea L.) growing in the Kemerovo region: functional foods application, Foods and Raw Materials, 2019, vol. 7, no. 1, pp. 84–92. https://doi.org/10.21603/2308-4057-2019-1-84-92

    Article  CAS  Google Scholar 

  20. Sadeghi Ekbatan, S., Li, X.Q., Ghorbani, M., et al., Chlorogenic acid and its microbial metabolites exert anti-proliferative effects, S-phase cell-cycle arrest and apoptosis in human colon cancer Caco-2 cells, Int. J. Mol. Sci., 2018, vol. 19, no. 3, art. ID 723. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5877584/#. Cited October 16, 2021. https://doi.org/10.3390/ijms19030723

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to A. Yu. Prosekov.

Ethics declarations

CONFLICT OF INTEREST

The authors declare that they have no conflicts of interest.

COMPLIANCE WITH ETHICAL STANDARDS

The study was performed without the use of animals or people as subjects.

Additional information

Translated by P. Kuchina

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Prosekov, A.Y., Kozlova, O.V. & Vesnina, A.D. Biotechnology of Cultivation of Rhaponticum Carthamoides (willd.) Suspension Cells: A Prospective Source of Antitumor Substances. Russ. Agricult. Sci. 48, 197–202 (2022). https://doi.org/10.3103/S1068367422030119

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S1068367422030119

Keywords:

Navigation