Synergistic effect of Zataria Multiflora essential oil on doxorubicin-induced growth inhibition of PC3 cancer cells and apoptosis
Introduction
Prostate cancer has a high prevalence among men worldwide, and it is the second cause of cancer-related death in the United States [1]. Prostate-specific antigen (PSA) screening, hormone therapy, and radiotherapy are used to treat prostate cancer. Although systemic chemotherapy is the alternative treatment when another treatment is not sufficient, unfortunately, these agents are relatively nonselective and highly toxic to normal tissues [2,3]. Doxorubicin belongs to anthracycline antibiotics family and is a useful type of chemotherapeutic agent with strong antineoplastic activity and is now widely used for many carcinomas and malignancies [4]. This antibiotic can causes DNA damage through insertion into the double helical structure of cancer cells DNA and free radical production by a redox reaction, and induce apoptotic cell death [5]. Although, doxorubicin is frequently used for prostate cancer treatment, due to its side effects such as immunosuppression and cardiomyopathy, and impact on normal cells its application faces dose limitations [3,6,7]. One strategy for decreasing the doxorubicin related systemic toxicity is combination treatment. Combination therapy is used to improve therapeutic efficacy, safety and reducing the side effects [3,8]. Natural compounds are potentials with high anticancer efficacy, and less toxicity to normal tissues and have been suggested as possible candidates to be examined for their synergistic effects in combination with anticancer drugs. Previous studies have shown that the use of natural and herbal compounds as combinatory therapy with doxorubicin for treating cancer has been successful [[9], [10], [11]]. Zataria Multiflora Boiss belongs to the Lamiaceae family and is widely distributed in Iran, Afghanistan, and Pakistan. Moreover, this plant with the local name of Avishan-e-Shirazi in Iran is an aromatic traditional medicinal plant. Zataria Multiflora essential oil (ZEO) is a hydrophobic concentrate that its hydrophobic nature allows it to readily cross the membrane to reach inside the cell [12]. As previous studies have indicated the major components of ZEO are Carvacrol, g-Terpinene, Carvacrol methyl ether, p-cymene and thymol [13,14]. ZEO due to its phenolic compounds has many biological properties including antioxidant, antibacterial and anti-inflammatory activities [15,16]. Studies have also shown that ZEO induces antiproliferative effect against MDA-MB-231 breast cancer cells without considerable cytotoxicity towards normal fibroblast cells [13]. For these reasons we have chosen ZEO to evaluate its synergistic effects in combination with doxorubicin to reduce the doxorubicin effective dosage and its nonspecific toxicity in a try to increase the efficacy of the therapy.
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Cell culture
PC3 cell line (a highly invasive human prostate cancer cell line) was purchased from National Cell Bank of Iran (Pasteur Institute, Iran) and was cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (Gibco) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin) and was incubated at 37 °C in a 5% CO2 atmosphere under 90–95% humidity.
Viability and proliferation assays
PC3 cells growth and viability were measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Sigma-Aldrich)
Antiproliferative effect of ZEO and doxorubicin on PC3 cells
In our previous study, we showed that ZEO has an antitumor effect on breast cancer cells while does not induce significant cytotoxic effect on L929 normal cells [13]. Here, ZEO with a dose range of 7.5–30 μg/ml inhibited the viability of PC3 cells with an IC50 value of 26.3 μg/ml after 48 h. Moreover, the IC50 value of doxorubicin was 1.89 μg/ml after 48 h (Fig. 1A). As shown in Fig. 1A, the cell viability decreased in a dose-dependent manner both in ZEO and doxorubicin treated PC3 cells. The
Discussion
One of the biggest challenges in the treatment of prostate cancer is lowering the toxicity of doxorubicin, a common anti-cancer drug. Pieces of evidence show that the natural compounds have been successful as combinatory therapy for cancer. In the present study, a drug co-administration therapeutic strategy was carried out with two compounds: one was the well-known doxorubicin with anticancer activity, and the other was ZEO, a polyphenolic Essential Oil isolated from Zataria Multiflora
CRediT authorship contribution statement
Elaheh Zare: Methodology, Validation, Formal Analysis, Investigation, Data Curation, Writing – Original Draft. Tahereh Jamali: Formal Analysis, Investigation, Writing – Review & Editing. Sussan K. Ardestani: Supervision, Project Administration, Funding Acquisition. Gholamreza Kavoosi: extracted and supplied the essential oil, Funding Acquisition.
Declaration of competing interest
The authors declare that they have no competing interest.
Acknowledgments
This study was jointly funded by the University of Tehran.
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