The Bioactive Properties of Carotenoids from Lipophilic Sea buckthorn Extract (Hippophae rhamnoides L.) in Breast Cancer Cell Lines

In women, breast cancer is the most commonly diagnosed cancer (11.7% of total cases) and the leading cause of cancer death (6.9%) worldwide. Bioactive dietary components such as Sea buckthorn berries are known for their high carotenoid content, which has been shown to possess anti-cancer properties. Considering the limited number of studies investigating the bioactive properties of carotenoids in breast cancer, the aim of this study was to investigate the antiproliferative, antioxidant, and proapoptotic properties of saponified lipophilic Sea buckthorn berries extract (LSBE) in two breast cancer cell lines with different phenotypes: T47D (ER+, PR+, HER2−) and BT-549 (ER-, PR-, HER2−). The antiproliferative effects of LSBE were evaluated by an Alamar Blue assay, the extracellular antioxidant capacity was evaluated through DPPH, ABTS, and FRAP assays, the intracellular antioxidant capacity was evaluated through a DCFDA assay, and the apoptosis rate was assessed by flow cytometry. LSBE inhibited the proliferation of breast cancer cells in a concentration-dependent manner, with a mean IC50 of 16 µM. LSBE has proven to be a good antioxidant both at the intracellular level, due to its ability to significantly decrease the ROS levels in both cell lines (p = 0.0279 for T47D, and p = 0.0188 for BT-549), and at the extracellular level, where the ABTS and DPPH inhibition vried between 3.38–56.8%, respectively 5.68–68.65%, and 35.6 mg/L equivalent ascorbic acid/g LSBE were recorded. Based on the results from the antioxidant assays, LSBE was found to have good antioxidant activity due to its rich carotenoid content. The flow cytometry results revealed that LSBE treatment induced significant alterations in late-stage apoptotic cells represented by 80.29% of T47D cells (p = 0.0119), and 40.6% of BT-549 cells (p = 0.0137). Considering the antiproliferative, antioxidant, and proapoptotic properties of the carotenoids from LSBE on breast cancer cells, further studies should investigate whether these bioactive dietary compounds could be used as nutraceuticals in breast cancer therapy.

[1]  M. Prieto,et al.  Valorization of Punica granatum L. Leaves Extracts as a Source of Bioactive Molecules , 2023, Pharmaceuticals.

[2]  Shuang Han,et al.  Targeting Mcl-1 Degradation by Bergenin Inhibits Tumorigenesis of Colorectal Cancer Cells , 2023, Pharmaceuticals.

[3]  N. Yaacob,et al.  Regulatory Mechanism on Anti-Glycolytic and Anti-Metastatic Activities Induced by Strobilanthes crispus in Breast Cancer, In Vitro , 2023, Pharmaceuticals.

[4]  Z. Cai,et al.  MAPK Cascade Signaling Is Involved in α-MMC Induced Growth Inhibition of Multiple Myeloma MM.1S Cells via G2 Arrest and Mitochondrial-Pathway-Dependent Apoptosis In Vitro , 2023, Pharmaceuticals.

[5]  P. Hrelia,et al.  In Vitro Investigation of the Anticancer Properties of Ammodaucus Leucotrichus Coss. & Dur. , 2022, Pharmaceuticals.

[6]  Yun Liu,et al.  Evaluation of the in vitro antioxidant and antitumor activity of extracts from Camellia fascicularis leaves , 2022, Frontiers in Chemistry.

[7]  A. Jemal,et al.  Breast Cancer Statistics, 2022 , 2022, CA: a cancer journal for clinicians.

[8]  Hamza Mechchate,et al.  LC–MS/MS Phytochemical Profiling, Antioxidant Activity, and Cytotoxicity of the Ethanolic Extract of Atriplex halimus L. against Breast Cancer Cell Lines: Computational Studies and Experimental Validation , 2022, Pharmaceuticals.

[9]  L. Mondello,et al.  Dittrichia viscosa L. Leaves: A Valuable Source of Bioactive Compounds with Multiple Pharmacological Effects , 2022, Molecules.

[10]  F. Cacciola,et al.  Beta vulgaris subsp. maritima: A Valuable Food with High Added Health Benefits , 2022, Applied Sciences.

[11]  H. Radeke,et al.  Antimicrobial activity, in vitro anticancer effect (MCF-7 breast cancer cell line), antiangiogenic and immunomodulatory potentials of Populus nigra L. buds extract , 2021, BMC Complementary Medicine and Therapies.

[12]  E. Gendaszewska-Darmach,et al.  Composition of flesh lipids and oleosome yield optimization of selected sea buckthorn (Hippophae rhamnoides L.) cultivars grown in Poland. , 2021, Food chemistry.

[13]  A. Diowksz,et al.  Wide Spectrum of Active Compounds in Sea Buckthorn (Hippophae rhamnoides) for Disease Prevention and Food Production , 2021, Antioxidants.

[14]  G. Borge,et al.  A comprehensive review on carotenoids in foods and feeds: status quo, applications, patents, and research needs , 2021, Critical reviews in food science and nutrition.

[15]  I. Berindan‐Neagoe,et al.  Zeaxanthin-Rich Extract from Superfood Lycium barbarum Selectively Modulates the Cellular Adhesion and MAPK Signaling in Melanoma versus Normal Skin Cells In Vitro , 2021, Molecules.

[16]  A. Stochmal,et al.  Novel bioactive properties of low-polarity fractions from sea-buckthorn extracts (Elaeagnus rhamnoides (L.) A. Nelson) - (in vitro). , 2020, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[17]  S. Wani,et al.  Sea buckthorn (Hippophae rhamnoides L.) inhibits cellular proliferation, wound healing and decreases expression of prostate specific antigen in prostate cancer cells in vitro , 2020 .

[18]  J. Nolan,et al.  From carotenoid intake to carotenoid blood and tissue concentrations – implications for dietary intake recommendations , 2020, Nutrition reviews.

[19]  M. Murkovic,et al.  Why is sea buckthorn (Hippophae rhamnoides L.) so exceptional? A review. , 2020, Food research international.

[20]  Xiang Xue,et al.  Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining. , 2020, Journal of visualized experiments : JoVE.

[21]  X. Bian,et al.  Triple-negative breast cancer molecular subtyping and treatment progress , 2020, Breast Cancer Research.

[22]  Y. Keum,et al.  Pro-oxidant Actions of Carotenoids in Triggering Apoptosis of Cancer Cells: A Review of Emerging Evidence , 2020, Antioxidants.

[23]  A. Urcan,et al.  Phytochemical Composition and Biological Activity of Berries and Leaves from Four Romanian Sea Buckthorn (Hippophae Rhamnoides L.) Varieties , 2020, Molecules.

[24]  Minhui Li,et al.  Advanced Research on the Antioxidant Activity and Mechanism of Polyphenols from Hippophae Species—A Review , 2020, Molecules.

[25]  T. Bohn,et al.  Sea Buckthorn Oil as a Valuable Source of Bioaccessible Xanthophylls , 2019, Nutrients.

[26]  Ł. Bobak,et al.  Anti-Oxidant and Anti-Enzymatic Activities of Sea Buckthorn (Hippophaë rhamnoides L.) Fruits Modulated by Chemical Components , 2019, Antioxidants.

[27]  M. Nhiri,et al.  Reactive Oxygen Species-Mediated Apoptosis and Cytotoxicity of Newly Synthesized Pyridazin-3-Ones In P815 (Murin Mastocytoma) Cell Line , 2019, Drug Research.

[28]  M. Stéger-Máté,et al.  Analysis of bioactive compounds of three sea buckthorn cultivars (Hippophaë rhamnoides L. ‘Askola’, ‘Leikora’, and ‘Orangeveja’) with HPLC and spectrophotometric methods , 2019, European Journal of Horticultural Science.

[29]  E. Fernandes,et al.  Antioxidant and pro-oxidant activities of carotenoids and their oxidation products. , 2018, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[30]  Kuan-rong Lee,et al.  Sulbactam-enhanced cytotoxicity of doxorubicin in breast cancer cells , 2018, Cancer cell international.

[31]  E. Rossi,et al.  Complementary and Integrative Medicine to Reduce Adverse Effects of Anticancer Therapy. , 2018, Journal of alternative and complementary medicine.

[32]  A. Wyss,et al.  Carotenoids in human nutrition and health. , 2018, Archives of biochemistry and biophysics.

[33]  V. Baskaran,et al.  Low-dose doxorubicin with carotenoids selectively alters redox status and upregulates oxidative stress-mediated apoptosis in breast cancer cells. , 2018, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[34]  Xiaoming Gong,et al.  Carotenoid Lutein Selectively Inhibits Breast Cancer Cell Growth and Potentiates the Effect of Chemotherapeutic Agents through ROS-Mediated Mechanisms , 2018, Molecules.

[35]  Changfu Zhu,et al.  A global perspective on carotenoids: Metabolism, biotechnology, and benefits for nutrition and health. , 2018, Progress in lipid research.

[36]  B. Olas,et al.  The Anticancer Activity of Sea Buckthorn [Elaeagnus rhamnoides (L.) A. Nelson] , 2018, Front. Pharmacol..

[37]  R. Liu,et al.  Phenolic compounds, antioxidant activity, antiproliferative activity and bioaccessibility of Sea buckthorn (Hippophaë rhamnoides L.) berries as affected by in vitro digestion. , 2017, Food & function.

[38]  V. Baskaran,et al.  Astaxanthin from shrimp efficiently modulates oxidative stress and allied cell death progression in MCF-7 cells treated synergistically with β-carotene and lutein from greens. , 2017, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[39]  R. Carle,et al.  Carotenoids and Carotenoid Esters of Red and Yellow Physalis (Physalis alkekengi L. and P. pubescens L.) Fruits and Calyces. , 2017, Journal of agricultural and food chemistry.

[40]  N. Chandel,et al.  The Two Faces of Reactive Oxygen Species in Cancer , 2017 .

[41]  R. Carle,et al.  Ultrastructural deposition forms and bioaccessibility of carotenoids and carotenoid esters from goji berries (Lycium barbarum L.). , 2017, Food chemistry.

[42]  V. Baskaran,et al.  Enhanced cytotoxic and apoptosis inducing activity of lycopene oxidation products in different cancer cell lines. , 2016, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[43]  M. Iranshahi,et al.  Prooxidant Activity of Polyphenols, Flavonoids, Anthocyanins and Carotenoids: Updated Review of Mechanisms and Catalyzing Metals , 2016, Phytotherapy research : PTR.

[44]  Zhen Huang,et al.  Supercritical CO2 extraction of Chinese lantern: Experimental and OEC modeling , 2016 .

[45]  A. Wojdyło,et al.  Analysis of Lipophilic and Hydrophilic Bioactive Compounds Content in Sea Buckthorn (Hippophaë rhamnoides L.) Berries. , 2015, Journal of agricultural and food chemistry.

[46]  J. Ouyang,et al.  Inhibitory effects of sea buckthorn procyanidins on fatty acid synthase and MDA-MB-231 cells , 2014, Tumor Biology.

[47]  J. Vincken,et al.  Carotenoid composition of berries and leaves from six Romanian sea buckthorn (Hippophae rhamnoides L.) varieties. , 2014, Food chemistry.

[48]  M. Valko,et al.  Health protective effects of carotenoids and their interactions with other biological antioxidants. , 2013, European journal of medicinal chemistry.

[49]  V. Böhm,et al.  Methods of measurement and evaluation of natural antioxidant capacity/activity (IUPAC Technical Report) , 2013 .

[50]  L. Mondello,et al.  Determination of carotenoids and their esters in fruits of sea buckthorn (Hippophae rhamnoides L.) by HPLC-DAD-APCI-MS. , 2012, Phytochemical analysis : PCA.

[51]  G. Floch,et al.  Antimicrobial, antioxidant and phytochemical investigations of sea buckthorn (Hippophaë rhamnoides L.) leaf, stem, root and seed , 2012 .

[52]  Takuji Tanaka,et al.  Cancer Chemoprevention by Caroteno , 2012, Molecules.

[53]  V. Böhm,et al.  Comparative antioxidant activities of carotenoids measured by ferric reducing antioxidant power (FRAP), ABTS bleaching assay (αTEAC), DPPH assay and peroxyl radical scavenging assay , 2011 .

[54]  Kil-Nam Kim,et al.  Fucoxanthin induces apoptosis in human leukemia HL-60 cells through a ROS-mediated Bcl-xL pathway. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[55]  P. Palozza.  Prooxidant actions of carotenoids in biologic systems. , 2009, Nutrition reviews.

[56]  E. Johansson,et al.  Carotenoids in sea buckthorn ( Hippophae rhamnoides L.) berries during ripening and use of pheophytin a as a maturity marker. , 2008, Journal of agricultural and food chemistry.

[57]  Abhishek Sharma,et al.  Microwave-assisted efficient extraction of different parts of Hippophae rhamnoides for the comparative evaluation of antioxidant activity and quantification of its phenolic constituents by reverse-phase high-performance liquid chromatography (RP-HPLC). , 2008, Journal of agricultural and food chemistry.

[58]  S. Elmore.  Apoptosis: A Review of Programmed Cell Death , 2007, Toxicologic pathology.

[59]  G. Blekas,et al.  Stability and radical-scavenging activity of heated olive oil and other vegetable oils , 2006 .

[60]  A. Varga,et al.  Chromatographic analysis of carotenol fatty acid esters in Physalis alkekengi and Hippophae rhamnoides. , 2005, Phytochemical analysis : PCA.

[61]  A. Nilsson,et al.  Inhibition of cancer cell proliferation in vitro by fruit and berry extracts and correlations with antioxidant levels. , 2004, Journal of agricultural and food chemistry.

[62]  T. G. Truscott,et al.  Carotenoid radical chemistry and antioxidant/pro-oxidant properties. , 2004, Archives of biochemistry and biophysics.

[63]  I. Wilson,et al.  Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. , 2000, European journal of biochemistry.

[64]  C. Rice-Evans,et al.  Antioxidant activity applying an improved ABTS radical cation decolorization assay. , 1999, Free radical biology & medicine.

[65]  T. G. Truscott,et al.  Prooxidant and antioxidant reaction mechanisms of carotene and radical interactions with vitamins E and C. , 1997, Nutrition.

[66]  J J Strain,et al.  The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. , 1996, Analytical biochemistry.

[67]  Chimedragchaa Chimedtseren,et al.  Traditional food, modern food and nutritional value of Sea buckthorn (Hippophae rhamnoides L.): a review , 2023, Journal of Future Foods.

[68]  V. Kuete,et al.  Anticancer Activities of African Medicinal Spices and Vegetables , 2017 .

[69]  C. D. Goldsmith,et al.  Fate of the phenolic compounds during olive oil production with the traditional press method , 2014 .

[70]  L. Surh,et al.  Prooxidant and Antioxidant Reaction Mechanisms of Carotene and Radical Interactions with Vitamins E and C , 1997 .