Skip to main content

Advertisement

SpringerLink
Log in

FNC (4′-azido-2′-deoxy-2′-fluoro(arbino)cytidine) as an Effective Therapeutic Agent for NHL: ROS Generation, Cell Cycle Arrest, and Mitochondrial-Mediated Apoptosis

  • Original Paper
  • Published:
Cell Biochemistry and Biophysics Aims and scope Submit manuscript

Abstract

Cytotoxic nucleoside analogs (NAs) hold great promise in cancer therapeutics by mimicking endogenous nucleosides and interfering with crucial cellular processes. Here, we investigate the potential of the novel cytidine analog, 4′-azido-2′-deoxy-2′-fluoro(arbino)cytidine (FNC), as a therapeutic agent for Non-Hodgkin lymphoma (NHL) using Dalton’s lymphoma (DL) as a T-cell lymphoma model. FNC demonstrated dose- and time-dependent inhibition of DL cell growth and proliferation. IC-50 values of FNC were measured at 1 µM, 0.5 µM, and 0.1 µM after 24, 48, and 72 h, respectively. Further elucidation of FNC’s mechanism of action uncovers its role in inducing apoptosis in DL cells. Notable DNA fragmentation and nuclear condensation point to activated apoptotic pathways. FNC-induced apoptosis was concomitant with changes in cellular membranes, characterized by membrane rupture and altered morphology. The robust anticancer effects of FNC are linked to its capacity to induce reactive oxygen species (ROS) production, prompting oxidative stress-mediated apoptosis. Additionally, FNC disrupted mitochondrial membrane potential (MMP), leading to mitochondrial dysfunction, further promoting apoptosis. Dysregulation of apoptotic genes, with upregulation of Bax and downregulation of Bcl-2 and Bcl-xl, implicates the mitochondrial-mediated apoptosis pathway. Furthermore, FNC-induced G2/M phase cell cycle arrest was mediated through modulation of the cell cycle inhibitor p21. Overall, this study highlights the potential of FNC as a promising therapeutic agent for NHL.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13

Similar content being viewed by others

Data Availability

This published article includes all of the data collected during this experimental study.

Abbreviations

AO:

Acridine Orange

DAPI:

4′,6-diamidino-2-phenylindole

DCFDA:

2’-7’-Dichlorodihydrofluorescein diacetate

DCT:

Dilution-cum-trypan assay

DL:

Dalton’s Lymphoma

EtBr:

Ethidium bromide

FNC:

4′-azido-2′-deoxy-2′-fluoro(arbino)cytidine

LA:

Late-apoptotic

MTT:

3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide

N:

Necrotic

NA:

Nucleoside analogue

NHL:

Non-Hodgkin lymphoma

PA:

Pre-apoptotic

PI:

Propidium iodide

Rh-123:

Rhodamine

ROS:

Reactive oxygen species

References

  1. Thandra, K. C., et al. (2021). Epidemiology of non-Hodgkin’s lymphoma. Medical Sciences, 9(1), 5.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Hjalgrim, H., & Engels, E. (2008). Infectious aetiology of Hodgkin and non‐Hodgkin lymphomas: a review of the epidemiological evidence. Journal of Internal Medicine, 264(6), 537–548.

    Article  CAS  PubMed  Google Scholar 

  3. De Jonghe, S., & Herdewijn, P. (2022). An Overview of Marketed Nucleoside and Nucleotide Analogs. Current Protocols, 2(3), e376.

    Article  PubMed  Google Scholar 

  4. Zhang, Y., et al. (2014). FNC, a novel nucleoside analogue, blocks invasion of aggressive non-Hodgkin lymphoma cell lines via inhibition of the Wnt/β-catenin signaling pathway. Asian Pacific Journal of Cancer Prevention, 15(16), 6829–6835.

    Article  PubMed  Google Scholar 

  5. Lee, S.-H., & Griffiths, J. R. (2020). How and why are cancers acidic? Carbonic anhydrase IX and the homeostatic control of tumour extracellular pH. Cancers, 12(6), 1616.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Yang, Q., et al. (2015). Synthesis and biological evaluation of 4-substituted fluoronucleoside analogs for the treatment of hepatitis B virus infection. Journal of Medicinal Chemistry, 58(9), 3693–3703.

    Article  CAS  PubMed  Google Scholar 

  7. Zhang, Y., et al. (2013). Effect of new nucleoside analogue FNC on proliferation, apoptosis and expressions of Bcl-6, PRDM1, C-myc in cell line Raji. J Zhengzhou University, 4, 450–454.

    Google Scholar 

  8. Zhang, Y., et al. (2017). FNC efficiently inhibits mantle cell lymphoma growth. PloS one, 12(3), e0174112.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Wang, Q., et al. (2011). FNC, a novel nucleoside analogue inhibits cell proliferation and tumor growth in a variety of human cancer cells. Biochemical pharmacology, 81(7), 848–855.

    Article  CAS  PubMed  Google Scholar 

  10. Faizullina, D., et al. (2022) FNC: an advanced anticancer therapeutic or just an underdog? Frontiers in Oncology, 198, 1–21.

  11. Singh, R. K., et al. (2021). Achyranthes aspera L. leaf extract induced anticancer effects on Dalton’s Lymphoma via regulation of PKCα signaling pathway and mitochondrial apoptosis. Journal of Ethnopharmacology, 274, 114060.

    Article  CAS  PubMed  Google Scholar 

  12. Kumar, N., et al. (2023). Safety Assessment of a Nucleoside Analogue FNC (2’-deoxy-2’-β-fluoro-4’-azidocytidine) in Balb/c Mice: Acute Toxicity Study. Asian Pacific Journal of Cancer Prevention: APJCP, 24(6), 2157–2170.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ribble, D., et al. (2005). A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnology, 5(1), 1–7.

    Article  Google Scholar 

  14. Chalut, K. J., et al. (2009). Light scattering measurements of subcellular structure provide noninvasive early detection of chemotherapy-induced apoptosis. Cancer Research, 69(3), 1199–1204.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Elkington, L., Adhikari, P., & Pradhan, P. (2022). Fractal Dimension Analysis to Detect the Progress of Cancer Using Transmission Optical Microscopy. Biophysica, 2(1), 59–69.

    Article  Google Scholar 

  16. Zaidieh, T., et al. (2019). ROS as a novel indicator to predict anticancer drug efficacy. BMC cancer, 19(1), 1–14.

    Article  Google Scholar 

  17. Gardiner, B., et al. (2020). Measurement of oxidative stress markers in vitro using commercially available kits. Measuring Oxidants and Oxidative Stress in Biological Systems, 39–60.

  18. Jeena, M., et al. (2019). Recent progress in mitochondria-targeted drug and drug-free agents for cancer therapy. Cancers, 12(1), 4.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Zorova, L. D., et al. (2018). Mitochondrial membrane potential. Analytical biochemistry, 552, 50–59.

    Article  CAS  PubMed  Google Scholar 

  20. Lapponi, M. J., et al. (2016). New developments in nucleoside analogues biosynthesis: A review. Journal of Molecular Catalysis B: Enzymatic, 133, 218–233.

    Article  CAS  Google Scholar 

  21. Asano, T. (2020). Drug resistance in cancer therapy and the possible role of epigenetics. Journal of Nippon Medical School, JNMS. 2020_87–508.

  22. Tsesmetzis, N., et al. (2018). Nucleobase and nucleoside analogues: resistance and re-sensitisation at the level of pharmacokinetics, pharmacodynamics and metabolism. Cancers, 10(7), 240.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Sun, L., et al. (2020). Mechanistic insight into antiretroviral potency of 2′-deoxy-2′-β-fluoro-4′-azidocytidine (FNC) with a long-lasting effect on HIV-1 prevention. Journal of Medicinal Chemistry, 63(15), 8554–8566.

    Article  CAS  PubMed  Google Scholar 

  24. Singh, R. K., et al. (2022). Evidence that PKCα inhibition in Dalton’s Lymphoma cells augments cell cycle arrest and mitochondrial-dependent apoptosis. Leukemia Research, 113, 106772.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

AA acknowledges SERB, IoE, and DHR for their grant. We are highly thankful to the ISLS, and CDC for providing the Fluorescence and Flow cytometry facility.

Author contributions

N.K.—Conceptualization, Methodology, Investigation, Writing - Original Draft, Writing-Review & Editing, Data analysis; A.S.—Methodology, Writing-Review & Editing; S.K.—Writing-Review & Editing; I.U.—Writing-Review & Editing; R.S.—Writing-Review & Editing; S.K.—Writing-Review & Editing; A.P.—Writing-Review & Editing; L.Y.—Writing- Review & Editing; R.T.—Writing- Review & Editing; R.P.—Writing-Review & Editing; S.P.M.—Writing-Review & Editing; K.—Writing-Review & Editing; J.A.—Writing-Review & Editing and Data analysis; A.A.—Writing-Original Draft, Writing-Review & Editing; Conceptualization, Supervision.

Funding

Preparation of this manuscript is funded by the University grant commission (UGC), New Delhi to NK in form of a Senior research fellowship (UGC Ref no.- 455/(CSIR-UGC NET DEC. 2017)). Prof Acharya is highly thankful to SERB, IoE, and DHR for their grant.

Author information

Authors and Affiliations

  1. Banaras Hindu University, Varanasi, India

    Naveen Kumar, Alok Shukla, Sanjay Kumar, Sandeep Kumar, Anand Patel, Lokesh Yadav, Ruchi Tiwari, Rachana Paswan, Shivashish Priyadarshi Mohanta & Arbind Acharya

  2. Sechenov First Moscow State Medical University, Moscow, Russia

    Ilya Ulasov

  3. Indian Institute of Technology, New Delhi, India

    Rishi Kant Singh

  4. Pachhunga University College Campus, Mizoram University, Aizawl, India

    Kaushalendra

  5. Lucknow University, Lucknow, India

    Jyeoti Antil

Authors
  1. Naveen Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alok Shukla
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sanjay Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ilya Ulasov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rishi Kant Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sandeep Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Anand Patel
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lokesh Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ruchi Tiwari
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Rachana Paswan
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Shivashish Priyadarshi Mohanta
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Kaushalendra
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Jyeoti Antil
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Arbind Acharya
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Arbind Acharya.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Ethical approval

The approval for the experiment conduction was taken from the Institutional Animal Ethics Committee of the Department of Zoology, Banaras Hindu University (BHU), Varanasi, India.

Inform consent

All authors have approved, agreed, and given consent to the submission of the manuscript.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kumar, N., Shukla, A., Kumar, S. et al. FNC (4′-azido-2′-deoxy-2′-fluoro(arbino)cytidine) as an Effective Therapeutic Agent for NHL: ROS Generation, Cell Cycle Arrest, and Mitochondrial-Mediated Apoptosis. Cell Biochem Biophys (2024). https://doi.org/10.1007/s12013-023-01193-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s12013-023-01193-6

Keywords

Search

Navigation