The anti-proliferative and apoptotic effects of crocin on chemosensitive and chemoresistant cervical cancer cells Homa Mollaei a,b , Reza Safaralizadeh a, *, Esmaeil Babaei a , Mohamad Reza Abedini b , Reyhane Hoshyar b, ** a Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran b Cellular & Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran A R T I C L E I N F O Article history: Received 11 March 2017 Received in revised form 6 July 2017 Accepted 11 July 2017 Keywords: Cervical cancer Crocin Chemoresistance Apoptosis MicroRNA A B S T R A C T Cervical cancer is the fourth cause of cancer-related mortality among females worldwide. Although current therapies reduce disease symptoms, resistance of tumor cells to chemotherapy agents after a while is a serious problem. Therefore, utilization of novel adjuvant agents to increase efficiency of chemotherapy is essential. In the last two decades, botanicals with effective anticancer activities have been studied. Among them, the anticancer properties of crocin have been more attended. In this study, the molecular mechanism of crocin action was investigated in sensitive human cervical cancer cell line (OV2008) in comparison with the resistant one (C13). A 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenylte- trazolium bromide (MTT) assay showed that crocin inhibits proliferation of sensitive cells (OV2008) at a time- and dose-dependent manner at 48 and 72 h. Also, this inhibitory effect has been shown on resistant cells (C13) at 72 h. Hoechst staining and flow cytometry assay also confirmed these results and revealed that antiproliferative effect of crocin might be due to the induction of apoptosis. Moreover, the genetic mechanism of crocin-induced apoptosis was accomplished by studying the relative expressions of P53, Bax, Bcl2 and miR-365, an upstream regulator of the last two ones. Real-time PCR analysis indicated that 1.5 and 3 mg/ml crocin led to up-regulation of Bax and P53 and down-regulation of Bcl2 and miR-365 at all time intervals in both two cell lines. However, OV2008 cell line was more sensitive to crocin, and alternation of gene expretion was more obvious in this cell line. In this regard, the present study demonstrated the anti-proliferative and apoptotic activities of crocin against both sensitive and resistant cervical cancer cells that may benefit cervical cancer treatment as an adjuvant agent to decrease chemoresistance and increase the efficiency of therapy. © 2017 Elsevier Masson SAS. All rights reserved. 1. Introduction Cervical cancer is one of the most common cancers among women worldwide [1,2]. Main therapeutic strategies for cervical cancer include surgery, radiotherapy, chemotherapy and combi- nation of them [3]. Although these therapies reduce disease symptoms, resistance of cancer cells to chemotherapy agents results in the recurrence of tumors [4]. To overcome this problem, researchers go finding adjuvant agents that improve effectiveness of current therapies. In recent decades, anticancer properties of herbal medicine have attracted scientists’ attention. The chemical analysis of Saffron (Crocus Sativus L.), a species belonging to the Iridaceae family [5], has shown the presence of more than 150 active metabolites in its stigmas [6]. Crocin, the main constituent of saffron extract, is one of the few water-soluble carotenoids in nature. Many studies have proved the anticancer effects of this carotenoid on various cancers invitro and invivo [7–10]. However, the exact mechanism of crocin effect is not fully known. Induction of apoptosis is one of the main molecular mechanisms of anticancer agents that can affect the chemoresistance of tumor cells too. Ability of tumor cells to escape from apoptosis signals has * Corresponding author at: Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, P.O. Box: 5166616471, Tabriz, Iran. ** Corresponding author at: Cellular & Molecular Research Center, Birjand University of Medical Sciences, P.O. Box: 9717853577, Birjand, Iran. E-mail addresses: safaralizadeh@tabrizu.ac.ir (R. Safaralizadeh), hooshyar@bums.ac.ir, reyhaneh.houshyar@gmail.com (R. Hoshyar). http://dx.doi.org/10.1016/j.biopha.2017.07.052 0753-3322/© 2017 Elsevier Masson SAS. All rights reserved. Biomedicine & Pharmacotherapy 94 (2017) 307–316 Available online at ScienceDirect www.sciencedirect.com