Evaluating the Cytotoxic and Apoptotic Potential of Terpinen-4-OLAgainst SF767 Glioblastoma Cells Via P53 Modulation: An In-Vitro Study

Authors

  • Neelam Bilal Author
  • Shazia Ramzan Author
  • Kanwar Sajjad Ali Author
  • Hina Sadaf Author
  • Zoja Muzaffar Author
  • Sehar Shamshad Ali Author
  • Zoraiz Chaudhry Author
  • Tahir Maqbool Author

DOI:

https://doi.org/10.59058/4vq08w26

Keywords:

Terpinen-4-ol (T-4-ol), Glioblastoma (GBM), apoptosis, MTTand Crystal Voilet assay

Abstract

Background and Objectives: Terpinen-4-ol (T4-ol) is a monoterpene derivative of essential oil which has
many anti- inflammatory, bactericidal and antifungal activities and is safe as compared to other treatment
options available. Our study's objectives were to analyze the effects of different concentrations of Terpinen4-ol on apoptosis of cell lines.
Methods: This in vitro experimental study was conducted at the IMBB Department, University of Lahore,
Lahore Pakistan, from October 2022 to March 2023. Study treatment was divided into four groups: negative
control (normal cell line), positive control (cisplatin) Terpinen-4-ol (experimental group) and vehicle control
(DMSO). In order to culture cells cryovials were thawed and then cells were cultured in T75 flasks. To assess
the cell viability MTT assay was used. Bioassay technology laboratory ELISA kit was used to evaluate
inflammation and apoptosis. Graph Pad prism was used to perform statistical analysis. The data was
analyzed using one- way analysis of variance (ANOVA) with bonferroni comparison within groups. P ≤ 0.05
was considered statistically significant.
Results: When compared to the control group, the treated lines showed more apoptotic changes. P-value was
≤ 0.05.
Conclusion: In this study we have observed anticancer effects of T4-ol against GBM cell line by observing
p53 induced apoptosis.
Key Words: Terpinen-4-ol (T-4-ol), Glioblastoma (GBM), apoptosis, MTTand Crystal Voilet assay.

 

References

1. Afify SM, Seno M. On the origin of cancer. Methods

in Cancer Stem Cell Biology: Springer; 2023. p. 1-

21

2. Fekete B. Survival of patients with glioblastomaadvancements in the past two decades2024.

3. Nguyen H, Guz-Montgomery K, Lowe D, Saha D.

Pathogenetic Features and Current Management of

Glioblastoma. Cancers 2021, 13, 856. 2021

4. Rajaratnam V, Islam MM, Yang M, Slaby R, Ramirez

HM, Mirza SP. Glioblastoma: Pathogenesis and

Current Status of Chemotherapy and Other Novel

Treatments. Cancers (Basel). 2020;12(4):937.10.-

3390/cancers12040937

5. Nguyen H-M, Guz-Montgomery K, Lowe DB, Saha

D. Pathogenetic features and current management of

glioblastoma. Cancers. 2021;13(4):856.

6. Brown NF, Ottaviani D, Tazare J, Gregson J, Kitchen

N, Brandner S, et al. Survival Outcomes and

Prognostic Factors in Glioblastoma. Cancers

(Basel). 2022;14(13):3161.10.3390/cancers1413-

3161

7. Morana O, Wood W, Gregory CD. The Apoptosis

Pa r a d o x i n Ca n c e r. I n t J M o l Sc i.

2022;23(3):1328.10.3390/ijms23031328

8. Nguyen TT, Wei S, Nguyen TH, Jo Y, Zhang Y, Park

W, et al. Mitochondria-associated programmed cell

death as a therapeutic target for age-related disease.

Ex p e rime n t a l & mo l e c u l a r me d i c i n e .

2023;55(8):1595-619.

9. Escobar E, Gomez-Valenzuela F, Penafiel C,

Chimenos-Kustner E, Perez-Tomas R. Aberrant

immunoexpression of p53 tumour-suppressor and

Bcl-2 family proteins (Bcl-2 and Bax) in

ameloblastomas and odontogenic keratocysts. J Clin

Exp Dent. 2023;15(2):e125e34.10.4317/jced.59769

10. Rizzotto D, Englmaier L, Villunger A. At a crossroads

to cancer: how p53-induced cell fate decisions

secure genome integrity. International journal of

molecular sciences. 2021;22(19):10883.

11. Shliapina VL, Yurtaeva SV, Rubtsova MP, Dontsova

OA. At the Crossroads: Mechanisms of Apoptosis

and Autophagy in Cell Life and Death. Acta

Na tur a e . 2021;13(2):106-15.10.32607/-

actanaturae.11208

12. Zhai K, Siddiqui M, Abdellatif B, Liskova A,

Kubatka P, Busselberg D. Natural Compounds in

Glioblastoma Therapy: Preclinical Insights,

Mechanistic Pathways, and Outlook. Cancers

(Basel). 2021;13(10):2317.10.3390/cancers131-

02317

13. Rong L, Li N, Zhang Z. Emerging therapies for

glioblastoma: current state and future directions. J

Exp Clin Cancer Res. 2022;41(1):142.10.1186/-

s13046-022-02349-7

14. Zimmermann RC, Poitevin CG, da Luz TS,

Mazarotto EJ, Furuie JL, Martins CEN, et al.

Antifungal activity of essential oils and their

combinations against storage fungi. Environmental

Science and Pollution Research. 2023;30(16)-

:48559-70.

15. Derbalah A, Shebl AM, Elgobashy SF, Ahmad AA,

Ramadan NE, Behiry SI, et al. Resistance induction

and direct antifungal activity of some monoterpenes

against Rhizoctonia solani, the causal of root rot in

common bean. Life. 2022;12(7):1040.

16. Deen JI, Zawad AS, Uddin M, Chowdhury MAH, Al

Araby SQ, Rahman MA. Terpinen-4-ol, A volatile

terpene molecule, extensively electrifies the

biological systems against the oxidative stresslinked pathogenesis. Advances in Redox Research.

2023;9:100082.

17. Chadha J, Khullar L, Mudgil U, Harjai K. A

comprehensive review on the pharmacological

prospects of Terpinen-4-ol: From nature to medicine

and beyond. Fitoterapia. 2024:106051.

18. Di Martile M, Garzoli S, Sabatino M, Valentini E,

D'Aguanno S, Ragno R, et al. Antitumor effect of

Melaleuca alternifolia essential oil and its main

component terpinen-4-ol in combination with target

therapy in melanoma models. Cell Death Discov.

2021;7(1):127.10.1038/s41420-021-00510-3

19. Maqbool T, Awan SJ, Malik S, Hadi F, Shehzadi S,

Tariq K. In-vitro anti-proliferative, apoptotic and

antioxidative activities of medicinal herb Kalonji

(Nigella sativa). Current pharmaceutical

biotechnology. 2019;20(15):1288-308.

20. Mitchell K, Troike K, Silver DJ, Lathia JD. The

evolution of the cancer stem cell state in

glioblastoma: emerging insights into the next

generation of functional interactions. Neurooncology. 2021;23(2):199-213.

21. Malik S, Awan SJ, Farzand A, Ali Q. Inflammation

reduction potential of nanostructured lipid carriers

encapsulated with rat's bone marrow cells' lysate.

Braz J Biol. 2023;82:e269553.10.1590/1519-

6984.269553

22. Upadhyay RK. Therapeutic and insecticidal potential

of plant terpenes: A review. International Journal of

Green Pharmacy (IJGP). 2022;16(1)

23. Cruz N, Herculano-Carvalho M, Roque D, Faria CC,

Cascão R, Ferreira HA, et al. Highlighted advances

in therapies for difficult-to-treat brain tumours such

as glioblastoma. Pharmaceutics. 2023;15(3):928.

24. Wu J, Li R, Wang J, Zhu H, Ma Y, You C, et al.

Reactive Astrocytes in Glioma: Emerging

Opportunities and Challenges. Int J Mol Sci.

2025;26(7):2907.10.3390/ijms26072907

25. Cao W, Tian R, Pan R, Sun B, Xiao C, Chen Y, et al.

Terpinen-4-ol inhibits the proliferation and mobility

of pancreatic cancer cells by downregulating Rhoassociated coiled-coil containing protein kinase 2.

Bioengineered. 2022;13(4):8643-56.10.1080/21-

655979.2022.2054205

26. Bozzuto G, Mariano F, Costa I, Calcabrini A,

Molinari A. Tea Tree Oil and Terpinen-4-Ol Induce

Cytoskeletal Reorganization of Human Melanoma

Cells. Planta Medica International Open.

2022;9(01):e34-e53.

27. Zhang Y, He L, Tu M, Huang M, Chen Y, Pan D, et al.

The ameliorative effect of terpinen-4-ol on ER

stress-induced vascular calcification depends on

SIRT1-mediated regulation of PERK acetylation.

Ph a rma c o l Re s. 2 0 2 1 ; 1 7 0 : 1 0 5 6 2 9 . 1 0 .-

1016/j.phrs.2021.105629

28. Casalle N, de Andrade CR. Cytotoxic and mutagenic

capacity of TTO and terpinen-4-ol in oral squamous

cell carcinoma. BioRxiv. 2020:2020.01. 03.893735.

29. Abid F, Saleem M, Muller CD, Asim MH, Arshad S,

Maqbool T, et al. Anti-proliferative and apoptosisinducing activity of Acacia Modesta and Opuntia

Monocantha extracts on HeLa cells. Asian Pacific

Journal of Cancer Prevention: APJCP.

2020;21(10):3125.

30. Rong D, Su Y, Zeng Z, Yang Y, Lu H, Cao YU.

Terpinen-4-ol suppresses proliferation and motility

of cutaneous squamous cell carcinoma cells by

enhancing calpain-2 expression. Oncol Res.

2025;33(3):605-16.10.32604/or.2024.050661

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Published

15-09-2025

How to Cite

Evaluating the Cytotoxic and Apoptotic Potential of Terpinen-4-OLAgainst SF767 Glioblastoma Cells Via P53 Modulation: An In-Vitro Study. (2025). JAIMC: Journal of Allama Iqbal Medical College, 23(2). https://doi.org/10.59058/4vq08w26