In vitro anticancer, antimicrobial and antifungal activity analysis of natural flavonoid hesperidin

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Berfu Çerçi Öngün
Ergül Mutlu Altundağ
Gülcem Altınoğlu
Mümtaz Güran
Gizem Şanlıtürk
Masoud Afshani
Deniz Balcı


Hesperidin, Anticancer, Anti-apoptosis, Antimicrobial, Antifungal


Hesperidin, a plant flavonoid mainly found in citrus fruits, has been shown to have important biological activities, while its effects on GBM cells and against pathogen microorganisms are largely unknown.  In this study, the cell viability effect of hesperidin on human GBM cells and the antibacterial and antifungal properties against a comprehensive set of microorganisms in vitro were explored. To assess the anti-proliferative effect of hesperidin on human GBM cells at different time points, GBM cells were treated with hesperidin at different concentrations. Cell viability was evaluated by MTT assay and cell nuclear morphology of hesperidin treated GBM cells was studied using DAPI staining. Hesperidin’s antimicrobial activity was screened with standard broth microdilution assays for antibacterial and antifungal activity screening. Antibacterial activity further tested by Antibacterial well diffusion assay. Hesperidin treatment significantly inhibited the cell viability in a concentration and time dependent manner, where the point of decrease in cell viability has been found at 150 and 200 μM of hesperidin treatment for 48 and 72 h. DAPI staining showed the induction of apoptosis in GBM cells following hesperidin treatment for 72 h at increased concentrations. The antimicrobial activity tests for hesperidin revealed no significant antimicrobial activity including antibacterial and antifungal assays. It can be concluded that hesperidin possesses anti-cancer properties in GBM cells in vitro resulting in reduced cell viability, proliferation and increased apoptosis, and may be effective against brain gliomas, with no significant antimicrobial activity.


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1. Wang TY, Li Q, Bi KS. Bioactive flavonoids in medicinal plants: Structure, activity and
biological fate. Asian J Pharm Sci 2018; 13(1): 12-23.
2. Krych J, Gebicka L. Catalase is inhibited by flavonoids. Int J Biol 2013; 58: 148-53.
3. Ragab FA, Yahya TAA, El-Naa MM, Arafa RK. Design, synthesis and structure–activity
relationship of novel semi-synthetic flavonoids as antiproliferative agents. Eur J Med Chem
2014; 82: 506-20.
4. Tian SS, Jiand FS, Zhang K, et al. Flavonoids from the leaves of Carya cathayensis Sarg.
inhibit vascular endothelial growth factor-induced angiogenesis. Fitoterapia 2014; 92: 34-
5. Willet WC. Balancing life-style and genomics research for disease prevention.
Science 2002; 296: 695-8.
6. Mahmoud AM, Ashour MB, Abdel-Moneim A, Ahmed OM. Hesperidin and naringin
attenuate hyperglycemia-mediated oxidative stress and proin ammatory cytokine
production in high fat fed/streptozotocin-induced type 2 diabetic rats. J Diabetes
Complications 2012; 26(6): 483-90.
7. SRKM Hewage, Piao MJ, Kang KA, et al. Hesperidin attenuates ultraviolet B-induced
apoptosis by mitigating oxidative stress in human keratinocytes. BiomolTher (Seoul) 2016;
24(3): 312-9.
8. Yvonne O, Driss F, Dang PMC, et al. Antioxidant effect of hydroxytyrosol, a polyphenol
from olive oil: scavenging of hydrogen peroxide but not superoxide anion produced by
human neutrophils. Biochem Pharmacol 2004; 68(10): 2003-8.
9. Afaq F, Saleem M, Krueger CG, Reed JD, Mukhtar H. Anthocyanin-and hydrolyzable
tannin-rich pomegranate fruit extract modulates MAPK and NF-B pathways and inhibits
skin tumorigenesis in CD-1 mice. Int J Cancer 2005; 113(3): 423-33.
10. Yang CS, Landau JM, Newmark HL. Inhibition of carcinogenesis by dietary polyphenolic
compounds. Annu Rev Nutr 2001; 21(1): 381-406.
11. Youdim KA, Dobbie MS, Kuhnle G, Proteggente AR, Abbott NJ, Rice-Evans C.
Interaction between flavonoids and the blood-brain barrier: in vitro studies. J Neurochem
2003; 85(1): 180-92.
12. Clinical and Laboratory Standards Institute. Reference method for broth dilution antifungal
susceptibility testing of yeasts, approved standard M27-A2. vol. 22, no. 15, 2nd ed., CLSI
Document. Clinical and Laboratory Standards Institute, Villanova, PA, 2002.
13. Haggag YA, El-Ashmawy NE, Okasha KM. Is hesperidin essential for prophylaxis and
treatment of COVID-19 Infection? Med Hypotheses 2020; 144.
14. Davis, ME. Glioblastoma: overview of disease and treatment. Clin J Oncol Nurs 2016;
20(5): 2-8.
15. Shu L, Cheung KL, Khor TO, Chen C, Kong AN. Phytochemicals: cancer chemoprevention
and suppression of tumor onset and metastasis. Cancer Metastasis Rev 2010; 29(3):
16. Nobili S, Lippi D, Witort E, et al. Natural compounds for cancer treatment and prevention.
Pharmacol Res 2009; 59(6): 365-78.
17. Brown JM, Attardi LD. The role of apoptosis in cancer development and treatment
response. Nat Rev Cancer 2005; 5(3): 231-7.
18. Hacker G. The morphology of apoptosis. Cell Tissue Res 2000; 301: 5-17.
19. Zhang XH, Zhang N, Meng X, Zhang Y, Qian Y, Xie YJ. Hesperetin inhibits the
proliferation of cerebrally implanted C6 glioma and involves suppression of HIF
1α/VEGF pathway in rats. Biomed Res 2017; 28(3).
20. Qiang L, Miao Z, Wang R, Yang J, Zhang D. Hesperetin Induces Apoptosis in Human
Glioblastoma Cells via p38 MAPK Activation. Nutr Cancer 2019; 72(3): 1-8.
21. Aggarwal V, Tuli HS, Thakral F, et al. Molecular mechanisms of action of hesperidin in
cancer: Recent trends and advancements. Exp Biol Med 2020; 245(5): 486-97.
22. Choi EJ. Hesperetin induced G1-phase cell cycle arrest in human breast cancer MCF-7
cells: involvement of CDK4 and p21. Nutr Cancer 2007; 59: 115–9.
23. Alshatwi AA, Ramesh E, Periasamy VS, Subash- Babu P. The apoptotic effect of
hesperetin on human cervical cancer cells is mediated through cell cycle arrest, death
receptor, and mitochondrial pathways. Fundam Clin Pharmacol 2013; 27: 581–92.
24. Palit S, Kar S, Sharma G, Das PK. Hesperetin induces apoptosis in breast carcinoma by
triggering accumulation of ROS and activation of ASK1/JNK pathway. J Cell Physiol
2015; 230:1729–39
25. Zhang J, Song J, Wu D, Wang J, Dong W. Hesperetin Induces the Apoptosis of
Hepatocellular Carcinoma Cells via Mitochondrial Pathway Mediated by the Increased
Intracellular Reactive Oxygen Species, ATP AND Calcium. Med Oncol 2015; 32: 101
26. Xia R, Sheng X, Xu X, Yu C, Lu H. Hesperidin induces apoptosis and
G0/G1 arrest in human non-small cell lung cancer A549 cells. Int J Mol
Med 2018; 41: 464–72
27. Birsu Cincin Z, Unlu M, Kiran B, Sinem Bireller E, Baran Y, Cakmakoglu B. Anti-
proliferative, apoptotic and signal transduction effects of hesperidin in non-small cell
lung cancer cells. Cell Oncol 2015; 38: 195–204.
28. Xie , Chen J, Xiao A, Liu, L. Antibacterial Activity of Polyphenols: Structure-Activity
Relationship and Influence of Hyperglycemic Condition. Molecules (Basel, Switzerland)
2017; 22(11): 1913.
29.Yang D, Wang T, Long M, Li P. Quercetin: Its Main Pharmacological Activity and Potential
Application in Clinical Medicine. Oxid Med Cell Longev 2020; 3: 1-13.
30. Bino A, Vicentini CB, Vertuani S, et al. Novel lipidized derivatives of the bioflavonoid
hesperidin: Dermatological, cosmetic and chemopreventive applications. Cosmetics 2018;
5(4): 72.
31. Güran M, Şanlıtürk G, Kerküklü NR, Altundağ EM, Süha Yalçın A. Combined effects of
quercetin and curcumin on anti-inflammatory and antimicrobial parameters in vitro. Eur J
Pharmacol 2019; 859.
32. Altundağ EM, Toprak K, Şanlıtürk G, et al. Synergistic Combination of Histone
Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid and Natural Flavonoid Curcumin
Exhibits Anticancer and Antibacterial Activity. Anticancer Agents Med Chem.

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