br a Research Team of
a Research Team of Cellular Genomics and Molecular Techniques of Investigations, Department of Biology, Faculty of Sciences, University of Moulay Ismail, B.P. 11201 Zitoune Meknès, Morocco
b UCIBIO.REQUIMTE, Laboratory of Biochemistry, Department of Biological Sciences, Faculty of Pharmacy, University of Porto, Rua Jorge Viterbo Ferreira, n° 228, 4050-313 Porto, Portugal c LAQV.REQUIMTE, Laboratory of Bromatology and Hydrology, Faculty of Pharmacy, University of Porto, Rua Jorge Viterbo Ferreira, n° 228, 4050-313 Porto, Portugal
Juniperus oxycedrus L.
Essential oils α-pinene β-myrcene
Fruits from Juniperus oxycedrus L. (Cupressaceae) are generally used in gastronomy and in folk medicine to treat cancer. In this study, the anti-cancer eﬃcacy of essential oils (EOs) of fruits and leaves from Moroccan J. oxy-cedrus L., was investigated in ER+ breast cancer cells. Both EOs presented anti-proliferative eﬀects but, cur-iously, only fruits EOs impaired cancer cell growth and induced apoptosis. To characterize the chemical com-position of these EOs, a suitable HS-SPME/GC–MS method was developed. Among the SPME fibers used, the PDMS and PA were the most eﬀective for characterization of volatile compounds. Moreover, these EOs are rich in α-pinene and β-myrcene, which also presented anti-cancer properties. For the first time, the anti-cancer properties of EOs and of new potential bioactive molecules of J. oxycedrus L. in ER+ breast cancer NPS-2143 were highlighted, suggesting their potential adjuvant therapeutic role in breast cancer.
Currently, Breast cancer is the second leading cancer that aﬀects women worldwide. Around 2.08 million new cancer cases were diag-nosed in 2018. This represents about 24.2% of all cancers in women and 11.6% of all new cancer cases (Bray, Ferlay, Soerjomataram, Siegel, Torre, & Jemal, 2018). From all breast cancer patients, around 70% of them have estrogen receptor-positive (ER+) breast cancer, which is a type of cancer that requires estrogens for growth. These hormones are
synthesized from androgens by aromatase, a product of the CYP19A1 gene and an enzyme that belongs to the superfamily of the cytochrome P450 enzymes (Chan, Petrossian, & Chen, 2016; Sobral, Amaral, Correia-da-Silva, & Teixeira, 2016). Aromatase inhibitors (AIs) and Selective estrogen receptor modulators (SERMs) are the therapeutic approaches used in clinic to treat this type of cancer. Unfortunately, despite their clinical eﬃcacy, these therapies may cause some adverse eﬀects like thromboembolism, bone fractures and lead to acquired re-sistance after prolonged treatment, which causes tumor re-growth
Abbreviations: A549 cells, Adenocarcinomic human alveolar basal epithelial cells; AIs, Aromatase inhibitors; ANOVA, One-Way Analyzes of Variance; CCCP, Carbonyl cyanide m-chlorophenylhydrazone; CFBS, Charcoal heat-inactivated fetal bovine serum; CYP19A1 gene, Cytochrome P450 family 19 subfamily A member 1 gene; DCFH2-DA, 2,7-dichlorodihydrofluorescein diacetate; DiOC6(3), 3,3′-dihexyloxacarbocyanine iodide probe; DMEM, Dulbecco’s Modified Eagle’s Medium; DMSO, Dimethyl sulfoxide; DPX, Slide mounting medium (polystyrene – plasticizer – xylene); DVB/CAR/PDMS, Divinylbenzene/carbonex/polydimethylsiloxane; Eos, Essential oils; ER+, Estrogen receptor-positive; FBS, Fetal bovine serum; GC–MS, Gas chromatography-mass spectrometry; HCA, Hierarchical cluster analyzes; HeLa, Human cervical carcinoma cell line; HepG2, Human liver cancer cell line; HFF-1, Human foreskin fibroblast cell line; HO-8910, Ovarian serous cystadeno-carcinoma cell line; HS-SPME, Headspace solid-phase microextraction; LDH, Lactate dehydrogenase; MCF-7, Breast cancer cell line (Michigan Cancer Foundation-7); MCF-7aro, ER+ aromatase-overexpressing human breast cancer cell line; MDA-MB-231, Breast adenocarcinoma cell line; MDA-MB-468, Mammary gland/breast cancer cell line; MEM, Minimum essential medium; MFI, Mean fluorescence intensity; MTT, 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide; PA, Polyacrylate; PBS, Phosphate-buﬀered saline; PCA, Principal component analyzes; PDMS, Poly-dimethyl-siloxane; PI, Propidium Iodide; PMA, Phorbol 12-myristate 13-acetate; RI, Retention indices; RLU, Relative luminescence units; ROS, Reactive oxygen species; SERMs, Selective estrogen receptor modulators; SK-OV-3, Ovarian serous cystadenocarcinoma cell line; STS, Staurosporine; T, Testosterone; UACC-257, Melanoma cell line; ψm, Mitochondrial transmembrane potential