Abstract
In this study, we compared the effects of terpinolene against the anti-cancer agent cisplatin on proliferation, apoptotic activity and intracellular ROS (reactive oxygen species) production in cancerous (MCF-7) and non-cancerous (HEK-293) cell lines. Cisplatin presented a strong growth-inhibitory effect on both MCF-7 (IC25: 18.77 µM) and HEK-293 (IC25: 15.03 µM) cell lines. The growth-inhibitory effect of terpinolene was found to be weaker than cisplatin (IC25: 291.18 µM for MCF-7 and 345.35 µM for HEK 293 cells). The expression levels of BAX (by 1.86-fold), cleaved-PARP (by 2.23-fold) and pro-caspase-8 (by 1.74-fold) proteins increased in response to terpinolene treatment in MCF-7 cells. Curiously, the increase in the expression levels of these apoptotic markers was less pronounced in terpinolene-treated non-cancerous HEK 293 cells. Terpinolene, promoted a higher rate of apoptotic and necrotic cell death on MCF-7 cells (apoptotic cells: 25.28 %; necrotic cells: 17.70 %) when compared with HEK-293 cell line (apoptotic cells: 15.47 %; necrotic cells: 9.42 %). Interestingly, terpinolene caused a marked increase (2.05-fold) in intracellular ROS production in MCF-7 cells while HEK-293 cells appeared to be resistant to terpinolene or cisplatin treatments. In conclusion, although terpinolene showed a weaker growth-inhibitory effect on MCF-7 cancer cells, it exhibited a better selectivity than cisplatin in inducing cell death and intracellular oxidative stress in MCF7 breast cancer cells.
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Data Availability
All the cell lines, raw experimental data, original images, data on statistical analysis and/or more detailed explanations on the material/methods section are available from the corresponding author on reasonable request.
Abbreviations
- ANOVA:
-
Analysis of variance
- BAX:
-
BCL2-associated X protein
- DCF-DA:
-
2’,7’-dichlorofluorescein-diacetate
- DMEM:
-
Dulbecco’s Modified Eagle Medium
- DMSO:
-
Dimethyl sulfoxid
- ECL:
-
Enhanced chemiluminescence
- EDTA:
-
Ethylenediamine tetraacetic acid FBS:Fetal Bovine Serum
- FITC:
-
Fluorescein isothiocyanate
- GAPDH:
-
Glyceraldehyde 3-phosphate dehydrogenase
- IC25 :
-
The concentration of the drug that is required for 25 % inhibition in vitro
- mTOR:
-
Mammalian Target of Rapamycin
- MTT:
-
[3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide]
- PARP:
-
Poly ADP (Adenosine Diphosphate)-Ribose Polymerase
- PBS:
-
Phosphate-buffered saline
- PBS-T:
-
Phosphate-buffered saline + Tween 20
- PI:
-
Propidium iodide
- P-S:
-
Penicillin and Streptomycin
- PVDF:
-
polyvinylidene difluoride
- ROS:
-
Reactive oxygen species
- SDS:
-
Sodium Dodecyl Sulfate
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Funding
This study was funded by the Turkish Scientific and Technical Research Council (TUBITAK) under contract no 118S206.
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Alper Calişkan and Elif Mertoglu carried out the experiments. Cenk Kig., Hizlan H. Agus and Evren O. Ucar supervised the project and contributed with critical reviews. Cenk Kig and Hizlan H. Agus designed the model of the study and provided literature search. Alper Caliskan, Elif Mertoglu and Varol Guler contributed in processing and analysis of data. Varol Guler contributed in the data collection and experimental studies during the project. Manuscript was written by Cenk Kig with contributions from Hizlan H. Agus and Evren O. Ucar.
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Kig, C., Mertoglu, E., Caliskan, A. et al. Selective and oxidative stress-mediated cell death of MCF-7 cell line induced by terpinolene. Biologia 76, 2757–2766 (2021). https://doi.org/10.1007/s11756-021-00803-z
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DOI: https://doi.org/10.1007/s11756-021-00803-z
Keywords
- Terpinolene
- MCF7
- HEK293
- apoptosis
- necrosis
- oxidative stress