Suppression of pulmonary tumor promotion and induction of apoptosis by Crocus sativus L. extraction
Authors: Saeed Samarghandian
Jalil Tavakkol Afshari
Saeideh Davoodi
Journal: Applied Biochemistry and Biotechnology (2011)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Crocus sativus L.; saffron; cytotoxic; apoptosis; carcinomic human alveolar basal epithelial cells
Affiliations: Department of Physiology, School of Medicine, Mashhad University Medical Sciences, Mashhad, Iran. 

Abstract:  Crocus sativus L., commonly known as saffron, is the raw material for one of the most expensive spice in the world, and it has been used in folk medicine for centuries. We investigated the potential of the ethanolic extract of saffron to induce cytotoxic and apoptosis effects in carcinomic human alveolar basal epithelial cells (A549), a commonly used cell culture system for in vitro studies on lung cancer. The cells were cultured in Dulbecco's modified Eagle's medium with 10% fetal bovine serum treated with different concentrations of the ethanolic extract of saffron for two consecutive days. Cell viability was quantitated by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Apoptotic cells were determined using annexin V-fluorescein isothiocyanate by flow cytometry. Saffron could decrease the cell viability in the malignant cells as a concentration- and time-dependent manner. The IC₅₀ values against the A549 cell lines were determined as 1,200 and 650 μg/ml after 24 and 48 h, respectively. Saffron-induced apoptosis of the A549 cells in a concentration-dependent manner, as determined by flow cytometry histogram of treated cells that induced apoptotic cell death, is involved in the toxicity of saffron. It might be concluded that saffron could cause cell death in the A549 cells, in which apoptosis plays an important role. Saffron could also be considered as a promising chemotherapeutic agent in lung cancer treatment in future. 

Retracted: No

PubPeer comments:
 can also be found in Fig 2(C) of [Samarghandian, Afshari Davoodi (2011)] , identified there as Fig 2 shows A549 cells after 24 hours of incubation with 400 u03bcg ml of saffron, and 48 h of incubation with 400 and 1,200 u03bcg ml of saffron. ![file] 2(C) also appears in Fig 3 of [Samarghandian, Boskabady Davoodi (2010)] , identified there as A549 cells after 24 hours of incubation with 500 u03bcg ml of saffron extract. ![file] , 
 #1 I will also go out on a limb to point out the similarities between Fig 2 and the celebrated Fig 2 of [Samarghandian et al (2013)] . ![file] ![file] , 
 *[left]* Fig 3.  Fig. 3 Malignant cells treated with concentrations of 400 and 1,200 u03bcg ml saffron for 48 h, the cells harvested, and apoptosis examined by flow cytometry . *[right]* Fig 3,  Assessment of apoptosis by Annexin-V PI on the renal cell carcinoma (ACHN). The cells were treated with different concentrations of honey (2.5% and 20%) for 48 h (II and III, respectively) or media (control symbol I), and apoptosis was examined with flow- cytometry after Annexin-V PI double staining . From  Honey induces apoptosis in renal cell carcinoma [(Samarghandian, Afshari, Davoodi 2011)] . ![file] Unexpected similarity between the effects of high concentrations of saffron and honey. , 
 Very similar patterns in two figures in a 2011 and 2013 paper: DOI: 10.1007 s12010-010-9130-x (2011; Springer Nature; Applied Biochemistry and Biotechnology; effect of saffron extract on lung cancer cells (A549)). Saeed Samarghandian, Jalil Tavakkol Afshari , Saeideh Davoodi. Suppression of pulmonary tumor promotion and induction of apoptosis by Crocus sativus L. extraction. Applied Biochemistry and Biotechnology (2011) 164:238u2013247 DOI: 10.1155 2013 417928 (2013; Hindawi; BioMed Research International; saffron effect on alveolar human lung cancer cells (A549)). Samarghandian S1, Borji A, Farahmand SK, Afshari R, Davoodi S. Crocus sativus L. (saffron) stigma aqueous extract induces apoptosis in alveolar human lung cancer cells through caspase-dependent pathways activation. Biomed Res Int. 2013;2013:417928. doi: 10.1155 2013 417928 https: imgur.com a dUDtc2c , 
 Now let s compare all three frames of Fig 3 (at left) with Fig 4 from [Samarghandian et al (2014)] (at right). ![file] , 
 In #5 above, the similarity between the panel III left and right goes further than outlined: the upper left quadrant would be the same except for a strange, sharp-edged rectangular gap where no points are present. A similar gap is seen in the upper right quadrant though a few points exist within it. , 
 Dear Sir or Madam, I am sorry for late, but I could not look the comments, since I could not open the file by our regular internet here. In fact, I have tried several ways for passing the internet problem here. But I could not open your file. Eventually, 2 days ago, a software company open it through a special software. Anyway, I am sorry for late reply. We would like to express our sincerely thank s for sending and inform us the comments. We are really sorry for any inconvenience for the comments. But, we really did not know anything about the comments and we do not have any idea how it is happened. We were looking for original data of those figures, but, we could not find it. Since it passed over than 10 years ago from getting the data and also our old laptop was broken (may it had some data). However, we can somehow remember, since the quality, size, resolution or clearness of our figures were low for publication in good journals, we gave the figures to a small company at that time for increasing the resolution and high quality, there is possibility during this process it may happened. We are not familiar to this kind of job. But again, we repeat, we do not know about it. We should inform you, the mentioned figures are resulted from the saffron effects on the A549 cells (lung cancer cell line) (by the MTT method) and its effects on cytotoxicity through Annexin V PI method. During 10 years ago, there are several published papers in very high rank Journals which confirm our mentioned data which published over than 10 years ago. The following published papers are some samples of those published papers. 1.tGezici S. Comparative anticancer activity analysis of saffron extracts and a principle component, crocetin for prevention and treatment of human malignancies. J Food Sci Technol. 2019 Dec;56(12):5435-5443. 2.tKhorasanchi Z, Shafiee M, Kermanshahi F, Khazaei M, Ryzhikov M, Parizadeh MR, Kermanshahi B, Ferns GA, Avan A, Hassanian SM. Crocus sativus a natural food coloring and flavoring has potent anti-tumor properties. Phytomedicine. 2018 Apr 1;43:21-27. 3.tGranchi C, Fortunato S, Meini S, Rizzolio F, Caligiuri I, Tuccinardi T, Lee HY, Hergenrother PJ, Minutolo F. Characterization of the Saffron Derivative Crocetin as an Inhibitor of Human Lactate Dehydrogenase 5 in the Antiglycolytic Approach against Cancer. J Agric Food Chem. 2017 Jul 19;65(28):5639-5649. 4.tChen S, Zhao S, Wang X, Zhang L, Jiang E, Gu Y, Shangguan AJ, Zhao H, Lv T, Yu Z. Crocin inhibits cell proliferation and enhances cisplatin and pemetrexed chemosensitivity in lung cancer cells. Transl Lung Cancer Res. 2015 Dec; 4(6):775-83. 5.tBolhassani A. Cancer chemoprevention by natural carotenoids as an efficient strategy. Anticancer Agents Med Chem. 2015;15(8):1026-31. 6.tLiu DD, Ye YL, Zhang J, Xu JN, Qian XD, Zhang Q. Distinct pro-apoptotic properties of Zhejiang saffron against human lung cancer via a caspase-8-9-3 cascade. Asian Pac J Cancer Prev. 2014;15(15):6075-80. 7.tBolhassani A, Khavari A, Bathaie SZ. Saffron and natural carotenoids: Biochemical activities and anti-tumor effects. Biochim Biophys Acta. 2014 Jan;1845 (1):20-30. 8.tZhong YJ, Shi F, Zheng XL, Wang Q, Yang L, Sun H, He F, Zhang L, Lin Y, Qin Y, Liao LC, Wang X. Crocetin induces cytotoxicity and enhances vincristine-induced cancer cell death via p53-dependent and -independent mechanisms. Acta Pharmacol Sin. 2011 Dec;32(12):1529-36. 9.tAbdullaev FI. Inhibitory effect of crocetin on intracellular nucleic acid and protein synthesis in malignant cells. Toxicol Lett. 1994 Feb 1;70(2):243-51. However, if it is needed, it is completely able to repeat them again in any experimental laboratories. Therefore, Although for many years, our research moved to animals studies and their laboratories, however, we can get those data again and again but it may need time to prepare the kits and materials for repeat experiment. Thank you very much for sending these comments again and sorry again for any inconvenience about it. Sincerely yours, Saeed Samarghandian ,