Assessment of antioxidant, antitumor and pro-apoptotic effects of Salvia fruticosa Mill. subsp. thomasii (Lacaita) Brullo, Guglielmo, Pavone & Terrasi (Lamiaceae) R. Tundis a, 1 , D. Iacopetta a, 1 , M.S. Sinicropi a, * , M. Bonesi a , M. Leporini a , N.G. Passalacqua b , J. Ceramella a , F. Menichini a , M.R. Loizzo a a Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, 87036 Rende, CS, Italy b Natural History Museum of Calabria and Botanic Garden, University of Calabria, I-87036 Rende, CS, Italy article info Article history: Received 22 February 2017 Received in revised form 27 April 2017 Accepted 19 May 2017 Available online 25 May 2017 Keywords: Salvia fruticosa Mill subsp. thomasii Antioxidant Antitumor Apoptosis abstract The aim of the present study was to investigate the in vitro antioxidant and antitumor effects of Salvia fruticosa Mill subsp. thomasii (Lacaita) Brullo, Guglielmo, Pavone & Terrasi (Lamiaceae). The aerial parts were extracted by maceration with methanol. This extract was partitioned with methanol and n-hexane. Luteolin, luteolin 7-O-glucoside, rutin and salvigenin were isolated from the methanol-soluble fraction. n-Hexane fraction showed viridiflorol, b-pinene, 1,8-cineole, as main components. The methanol-soluble fraction exerted antitumor activity against human breast cancer (MCF-7 and MDA-MB-231) and human colorectal carcinoma (RKO and Caco-2) cells. TUNEL test revealed that S. fruticosa subsp. thomasii leads to cells death by apoptosis, with low cytotoxic effects on non-tumoral 3T3-L1 cells. Moreover, it exerted the highest protection of lipid peroxidation and reduced the oxidative stress induced by menadione treat- ment in 3T3-L1 murine fibroblasts. S. fruticosa subsp. thomasii bioactivity could promote its use not only as food but also in nutraceutical/pharmaceutical industries. © 2017 Elsevier Ltd. All rights reserved. 1. Introduction According to World Cancer Report (WHO) Cancer rates could further increase by 50% to 15 million new cases in the year 2020 (http://www.who.int/whr/en/). The most common causes of death are lung cancer (1.6 million deaths), liver cancer (745,000 deaths), and stomach cancer (723,000 deaths). By 2030, it is projected that there will be 26 million new cancer cases and 17 million cancer deaths per year. Plants have been used for medical purposes since the beginning of human history and may be considered the basis of modern medicine (Newman and Cragg, 2016; Parisi et al., 2013; Sala et al., 2013). Many natural products and compounds derived by plants (Chimento et al., 2013; Chosson et al., 2012; da Rocha et al., 2001; Saturnino et al., 2015; Sinicropi et al., 2009) have received increasing attention, over the past 30 years, for their potential as novel cancer preventive and therapeutic agents. Moreover, any evidences demonstrating the potential of plant-derived com- pounds as inhibitors of various stages of tumorigenesis and asso- ciated inflammatory and oxidant processes have been published underlying the importance of these products in cancer prevention Abbreviations: ACHN, renal adenocarcinoma cell line; DPPH, 2,2-diphenyl-1- picrylhydrazyl; FRAP, Ferric Reducing Ability Power; ROS, Intracellular Reactive Oxygen Species; DCFH-DA, dichloro-dihydro-fluorescein diacetate; MTT, 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; TUNEL, Terminal deoxy- nucleotidyl transferase dUTP nick end labeling; TLC, thin layer chromatography; TPTZ, tripyridyltriazine; BHT, butylated hydroxytoluene; ER, estrogen receptor; NCS, Newborn Calf Serum; FBS, Fetal Bovine Serum; MPLC, Medium Pressure Liquid Chromatography; GC, Gas Chromatography; GC-MS, Gas Chromatography-Mass Spectrometry; FID, flame-ionization detector; NAC, N-acetylcysteine; Men, mena- dione; PBF, phosphate-buffered saline; DAPI, 2-(4-amidinophenyl)-6- indolecarbamidine dihydrochloride; TdT, terminal deoxynucleotidyl transferase; SD, standard deviation; UPLC, ultraperformance liquid chromatography; MS, mass spectrometry; NF-kB, nuclear factor kappa B; PI3K, phosphatidylinositol 3 0 -kinase; XIAP, X-linked inhibitor of apoptosis protein; MYCN, N-myc proto-oncogene pro- tein; TNF-a, Tumor Necrosis Factor-alpha; MAPK, Mitogen Activated Protein Kinase; ERK, extracellular signaleregulated kinase. * Corresponding author. E-mail addresses: rosa.tundis@unical.it (R. Tundis), domenico.iacopetta@unical. it (D. Iacopetta), s.sinicropi@unical.it (M.S. Sinicropi), marco.bonesi@unical.it (M. Bonesi), mariarosarialeporini@tiscali.it (M. Leporini), nicodemo.passalacqua@ unical.it (N.G. Passalacqua), francesco.menichini@unical.it (F. Menichini), monica_ rosa.loizzo@unical.it (M.R. Loizzo). 1 These authors have contributed equally to the manuscript. Contents lists available at ScienceDirect Food and Chemical Toxicology journal homepage: www.elsevier.com/locate/foodchemtox http://dx.doi.org/10.1016/j.fct.2017.05.040 0278-6915/© 2017 Elsevier Ltd. All rights reserved. Food and Chemical Toxicology 106 (2017) 155e164