BT-474 Breast Cancer Cell Apoptosis Induced by Crocin, a Saffron Carotenoid
Abstract
Background: Saffron carotenoids have been known as powerful phytochemicals in breast cancer treatment. The purpose of this study was to investigate the anti-cancer properties of an important saffron carotenoid, crocin, on BT-474 which is a known HER2+ breast cancer cell line. Methods: The effect of crocin on cell viability was investigated using MTT assay. Apoptosis induction was studied via flow cytometry and Western blotting of caspase-9 and cleaved caspase-9. Involvement of unfolded protein response (UPR) was also investigated via RT-PCR study of the XBP1 gene. Results: The results showed that crocin treatment decreases the viability of BT-474 cells and induces early and late apoptosis in these cells. The mechanism of crocin action was through the induction of caspase-9 expression and cleavage. Furthermore, we found that crocin induced XBP1 gene splicing in these cells. Conclusion: The present study provides important evidence that crocin induces apoptosis in BT-474 cells. In addition, the activation of UPR may play a role in the anticancer effects of crocin.
1. Mousavi SM, Montazeri A, Mohagheghi MA, Jarrahi AM, Harirchi I, Najafi M, et al. Breast cancer in Iran: an epidemiological review. Breast J. 2007;13(4):383-91.
2. M. N. Mortality views in 18 Provinces of Iran 2001. Ministry of Health, Deputy to Health Directory, Research and development office. 2003;75.
3. Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin. 2010;60(5):277-300.
4. Rotilio G, Mavelli I, Rossi L, Ciriolo MR. Biochemical mechanism of oxidative damage by redox-cycling drugs. Environ Health Perspect. 1985;64:259-64.
5. Kamesaki H. Mechanisms involved in chemotherapy-induced apoptosis and their implications in cancer chemotherapy. Int J Hematol. 1998;68(1):29-43.
6. Hynes NE, Stern DF. The biology of erbB-2/neu/HER-2 and its role in cancer. Biochim Biophys Acta. 1994;1198(2-3):165-84.
7. Bartlett JM, Going JJ, Mallon EA, Watters AD, Reeves JR, Stanton P, et al. Evaluating HER2 amplification and overexpression in breast cancer. J Pathol. 2001;195(4):422-8.
8. Lipton A, Ali SM, Leitzel K, Demers L, Chinchilli V, Engle L, et al. Elevated serum Her-2/neu level predicts decreased response to hormone therapy in metastatic breast cancer. J Clin Oncol. 2002;20(6):1467-72.
9. Subramaniam DS, Isaacs C. Utilizing prognostic and predictive factors in breast cancer. Curr Treat Options Oncol. 2005;6(2):147-59.
10. Pasleau F, Grooteclaes M, Gol-Winkler R. Expression of the c-erbB2 gene in the BT474 human mammary tumor cell line: measurement of c-erbB2 mRNA half-life. Oncogene. 1993;8(4):849-54.
11. Schneider Y, Vincent F, Duranton B, Badolo L, Gosse F, Bergmann C, et al. Anti-proliferative effect of resveratrol, a natural component of grapes and wine, on human colonic cancer cells. Cancer Lett. 2000;158(1):85-91.
12. Ueda S, Masutani H, Nakamura H, Tanaka T, Ueno M, Yodoi J. Redox control of cell death. Antioxid Redox Signal. 2002;4(3):405-14.
13. Galluzzi L, Vitale I, Abrams JM, Alnemri ES, Baehrecke EH, Blagosklonny MV, et al. Molecular definitions of cell death subroutines: recommendations of the Nomenclature Committee on Cell Death 2012. Cell Death Differ. 2012;19(1):107-20.
14. Taylor RC, Cullen SP, Martin SJ. Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol. 2008;9(3):231-41.
15. Kidd VJ. Proteolytic activities that mediate apoptosis. Annu Rev Physiol. 1998;60:533-73.
16. Krajewski S, Krajewska M, Ellerby LM, Welsh K, Xie Z, Deveraux QL, et al. Release of caspase-9 from mitochondria during neuronal apoptosis and cerebral ischemia. Proc Natl Acad Sci U S A. 1999;96(10):5752-7.
17. Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, et al. Mitochondrial release of caspase-2 and -9 during the apoptotic process. J Exp Med. 1999;189(2):381-94.
18. Uemura A, Oku M, Mori K, Yoshida H. Unconventional splicing of XBP1 mRNA occurs in the cytoplasm during the mammalian unfolded protein response. J Cell Sci. 2009;122(Pt 16):2877-86.
19. Cawley K, Deegan S, Samali A, Gupta S. Assays for detecting the unfolded protein response. Methods Enzymol. 2011;490:31-51.
20. Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K. XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell. 2001;107(7):881-91.
21. Sovolyova N, Healy S, Samali A, Logue SE. Stressed to death - mechanisms of ER stress-induced cell death. Biological chemistry. 2014;395(1):1-13.
22. Hu R, Warri A, Jin L, Zwart A, Riggins RB, Fang HB, et al. NF-kappaB signaling is required for XBP1 (unspliced and spliced)-mediated effects on antiestrogen responsiveness and cell fate decisions in breast cancer. Molecular and cellular biology. 2015;35(2):379-90.
23. Clarke R, Cook KL, Hu R, Facey CO, Tavassoly I, Schwartz JL, et al. Endoplasmic reticulum stress, the unfolded protein response, autophagy, and the integrated regulation of breast cancer cell fate. Cancer research. 2012;72(6):1321-31.
24. Verfaillie T, Salazar M, Velasco G, Agostinis P. Linking ER Stress to Autophagy: Potential Implications for Cancer Therapy. International journal of cell biology. 2010;2010:930509.
25. Bathaie SZ, Mousavi SZ. New applications and mechanisms of action of saffron and its important ingredients. Crit Rev Food Sci Nutr. 2010;50(8):761-86.
26. Mousavi SZ, Bathaie SZ. Historical uses of saffron: Identifying potential new avenues for modern research. Avicenna Journal of Phytomedicine. 2011;1(2):57-66.
27. Soeda S, Ochiai T, Shimeno H, Saito H, Abe K, Tanaka H, et al. Pharmacological activities of crocin in saffron. Journal of Natural Medicines. 2007;61(2):102-11.
28. Escribano J, Alonso GL, Coca-Prados M, Fernandez JA. Crocin, safranal and picrocrocin from saffron (Crocus sativus L.) inhibit the growth of human cancer cells in vitro. Cancer Lett. 1996;100(1-2):23-30.
29. Zhao P, Luo CL, Wu XH, Hu HB, Lv CF, Ji HY. [Proliferation apoptotic influence of crocin on human bladder cancer T24 cell line]. Zhongguo Zhong Yao Za Zhi. 2008;33(15):1869-73.
30. Ashrafi M, Bathaie SZ, Abroun S, Azizian M. Effect of Crocin on Cell Cycle Regulators in N-Nitroso-N-Methylurea-Induced Breast Cancer in Rats. DNA Cell Biol. 2015;34(11):684-91.
31. Hoshyar R, Bathaie SZ, Sadeghizadeh M. Crocin triggers the apoptosis through increasing the Bax/Bcl-2 ratio and caspase activation in human gastric adenocarcinoma, AGS, cells. DNA Cell Biol. 2013;32(2):50-7.
32. Hoshyar R, Bathaie SZ, Kyani A, Mousavi MF. Is there any interaction between telomeric DNA structures, G-quadruplex and I-motif, with saffron active metabolites? Nucleosides Nucleotides Nucleic Acids. 2012;31(11):801-12.
33. Ashrafi M, Bathaie SZ, Taghikhani M, Moosavi-Movahedi AA. The effect of carotenoids obtained from saffron on histone H1 structure and H1-DNA interaction. Int J Biol Macromol. 2005;36(4):246-52.
34. Bathaie SZ, Bolhasani A, Hoshyar R, Ranjbar B, Sabouni F, Moosavi-Movahedi AA. Interaction of saffron carotenoids as anticancer compounds with ctDNA, Oligo (dG.dC)15, and Oligo (dA.dT)15. DNA Cell Biol. 2007;26(8):533-40.
35. Bolhasani A, Bathaie S, Yavari I, Moosavi-Movahedi A, Ghaffari M. Separation and purification of some components of Iranian saffron. Asian Journal of Chemistry. 2005;17(2):725.
36. Denis L, Morton MS, Griffiths K. Diet and its preventive role in prostatic disease. Eur Urol. 1999;35(5-6):377-87.
37. Bathaie SZ, Bolhassani A, Tamanoi F. Anticancer Effect and Molecular Targets of Saffron Carotenoids. Enzymes. 2014;36:57-86.
38. D'Alessandro AM, Mancini A, Lizzi AR, De Simone A, Marroccella CE, Gravina GL, et al. Crocus sativus stigma extract and its major constituent crocin possess significant antiproliferative properties against human prostate cancer. Nutr Cancer. 2013;65(6):930-42.
39. Mousavi B, Bathaie SZ, Fadai F, Ashtari Z, Ali Beigi N, Farhang S, et al. Safety evaluation of saffron stigma (Crocus sativus L.) aqueous extract and crocin in patients with schizophrenia. Avicenna J Phytomed. 2015;5(5):413-9.
40. Heidarzadeh H, Bathaie SZ, Abroun S, Mohagheghi MA. Evaluating the cytotoxic effect of crocin on MDA-MB-468 cell line based on apoptosis induction, ER stress, and autophagy markers. Pathobiology Research. 2018;20(4):37-51.
41. Debatin KM, Poncet D, Kroemer G. Chemotherapy: targeting the mitochondrial cell death pathway. Oncogene. 2002;21(57):8786-803.
42. Fulda S. Modulation of apoptosis by natural products for cancer therapy. Planta Med. 2010;76(11):1075-9.
43. Fesik SW. Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer. 2005;5(11):876-85.
44. Salih SM, Ringelstetter AK, Elsarrag MZ, Abbott DH, Roti EC. Dexrazoxane abrogates acute doxorubicin toxicity in marmoset ovary. Biol Reprod. 2015;92(3):73.
45. Bakshi HA, Hakkim FL, Sam S. Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation. Asian Pac J Cancer Prev. 2016;17(3):1499-506.
46. Mousavi SH, Tayarani NZ, Parsaee H. Protective effect of saffron extract and crocin on reactive oxygen species-mediated high glucose-induced toxicity in PC12 cells. Cell Mol Neurobiol. 2010;30(2):185-91.
47. Thomas E, Gopalakrishnan V, Somasagara RR, Choudhary B, Raghavan SC. Extract of Vernonia condensata, Inhibits Tumor Progression and Improves Survival of Tumor-allograft Bearing Mouse. Scientific reports. 2016;6:23255.
48. Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol. 2012;13(2):89-102.
2. M. N. Mortality views in 18 Provinces of Iran 2001. Ministry of Health, Deputy to Health Directory, Research and development office. 2003;75.
3. Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin. 2010;60(5):277-300.
4. Rotilio G, Mavelli I, Rossi L, Ciriolo MR. Biochemical mechanism of oxidative damage by redox-cycling drugs. Environ Health Perspect. 1985;64:259-64.
5. Kamesaki H. Mechanisms involved in chemotherapy-induced apoptosis and their implications in cancer chemotherapy. Int J Hematol. 1998;68(1):29-43.
6. Hynes NE, Stern DF. The biology of erbB-2/neu/HER-2 and its role in cancer. Biochim Biophys Acta. 1994;1198(2-3):165-84.
7. Bartlett JM, Going JJ, Mallon EA, Watters AD, Reeves JR, Stanton P, et al. Evaluating HER2 amplification and overexpression in breast cancer. J Pathol. 2001;195(4):422-8.
8. Lipton A, Ali SM, Leitzel K, Demers L, Chinchilli V, Engle L, et al. Elevated serum Her-2/neu level predicts decreased response to hormone therapy in metastatic breast cancer. J Clin Oncol. 2002;20(6):1467-72.
9. Subramaniam DS, Isaacs C. Utilizing prognostic and predictive factors in breast cancer. Curr Treat Options Oncol. 2005;6(2):147-59.
10. Pasleau F, Grooteclaes M, Gol-Winkler R. Expression of the c-erbB2 gene in the BT474 human mammary tumor cell line: measurement of c-erbB2 mRNA half-life. Oncogene. 1993;8(4):849-54.
11. Schneider Y, Vincent F, Duranton B, Badolo L, Gosse F, Bergmann C, et al. Anti-proliferative effect of resveratrol, a natural component of grapes and wine, on human colonic cancer cells. Cancer Lett. 2000;158(1):85-91.
12. Ueda S, Masutani H, Nakamura H, Tanaka T, Ueno M, Yodoi J. Redox control of cell death. Antioxid Redox Signal. 2002;4(3):405-14.
13. Galluzzi L, Vitale I, Abrams JM, Alnemri ES, Baehrecke EH, Blagosklonny MV, et al. Molecular definitions of cell death subroutines: recommendations of the Nomenclature Committee on Cell Death 2012. Cell Death Differ. 2012;19(1):107-20.
14. Taylor RC, Cullen SP, Martin SJ. Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol. 2008;9(3):231-41.
15. Kidd VJ. Proteolytic activities that mediate apoptosis. Annu Rev Physiol. 1998;60:533-73.
16. Krajewski S, Krajewska M, Ellerby LM, Welsh K, Xie Z, Deveraux QL, et al. Release of caspase-9 from mitochondria during neuronal apoptosis and cerebral ischemia. Proc Natl Acad Sci U S A. 1999;96(10):5752-7.
17. Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, et al. Mitochondrial release of caspase-2 and -9 during the apoptotic process. J Exp Med. 1999;189(2):381-94.
18. Uemura A, Oku M, Mori K, Yoshida H. Unconventional splicing of XBP1 mRNA occurs in the cytoplasm during the mammalian unfolded protein response. J Cell Sci. 2009;122(Pt 16):2877-86.
19. Cawley K, Deegan S, Samali A, Gupta S. Assays for detecting the unfolded protein response. Methods Enzymol. 2011;490:31-51.
20. Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K. XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell. 2001;107(7):881-91.
21. Sovolyova N, Healy S, Samali A, Logue SE. Stressed to death - mechanisms of ER stress-induced cell death. Biological chemistry. 2014;395(1):1-13.
22. Hu R, Warri A, Jin L, Zwart A, Riggins RB, Fang HB, et al. NF-kappaB signaling is required for XBP1 (unspliced and spliced)-mediated effects on antiestrogen responsiveness and cell fate decisions in breast cancer. Molecular and cellular biology. 2015;35(2):379-90.
23. Clarke R, Cook KL, Hu R, Facey CO, Tavassoly I, Schwartz JL, et al. Endoplasmic reticulum stress, the unfolded protein response, autophagy, and the integrated regulation of breast cancer cell fate. Cancer research. 2012;72(6):1321-31.
24. Verfaillie T, Salazar M, Velasco G, Agostinis P. Linking ER Stress to Autophagy: Potential Implications for Cancer Therapy. International journal of cell biology. 2010;2010:930509.
25. Bathaie SZ, Mousavi SZ. New applications and mechanisms of action of saffron and its important ingredients. Crit Rev Food Sci Nutr. 2010;50(8):761-86.
26. Mousavi SZ, Bathaie SZ. Historical uses of saffron: Identifying potential new avenues for modern research. Avicenna Journal of Phytomedicine. 2011;1(2):57-66.
27. Soeda S, Ochiai T, Shimeno H, Saito H, Abe K, Tanaka H, et al. Pharmacological activities of crocin in saffron. Journal of Natural Medicines. 2007;61(2):102-11.
28. Escribano J, Alonso GL, Coca-Prados M, Fernandez JA. Crocin, safranal and picrocrocin from saffron (Crocus sativus L.) inhibit the growth of human cancer cells in vitro. Cancer Lett. 1996;100(1-2):23-30.
29. Zhao P, Luo CL, Wu XH, Hu HB, Lv CF, Ji HY. [Proliferation apoptotic influence of crocin on human bladder cancer T24 cell line]. Zhongguo Zhong Yao Za Zhi. 2008;33(15):1869-73.
30. Ashrafi M, Bathaie SZ, Abroun S, Azizian M. Effect of Crocin on Cell Cycle Regulators in N-Nitroso-N-Methylurea-Induced Breast Cancer in Rats. DNA Cell Biol. 2015;34(11):684-91.
31. Hoshyar R, Bathaie SZ, Sadeghizadeh M. Crocin triggers the apoptosis through increasing the Bax/Bcl-2 ratio and caspase activation in human gastric adenocarcinoma, AGS, cells. DNA Cell Biol. 2013;32(2):50-7.
32. Hoshyar R, Bathaie SZ, Kyani A, Mousavi MF. Is there any interaction between telomeric DNA structures, G-quadruplex and I-motif, with saffron active metabolites? Nucleosides Nucleotides Nucleic Acids. 2012;31(11):801-12.
33. Ashrafi M, Bathaie SZ, Taghikhani M, Moosavi-Movahedi AA. The effect of carotenoids obtained from saffron on histone H1 structure and H1-DNA interaction. Int J Biol Macromol. 2005;36(4):246-52.
34. Bathaie SZ, Bolhasani A, Hoshyar R, Ranjbar B, Sabouni F, Moosavi-Movahedi AA. Interaction of saffron carotenoids as anticancer compounds with ctDNA, Oligo (dG.dC)15, and Oligo (dA.dT)15. DNA Cell Biol. 2007;26(8):533-40.
35. Bolhasani A, Bathaie S, Yavari I, Moosavi-Movahedi A, Ghaffari M. Separation and purification of some components of Iranian saffron. Asian Journal of Chemistry. 2005;17(2):725.
36. Denis L, Morton MS, Griffiths K. Diet and its preventive role in prostatic disease. Eur Urol. 1999;35(5-6):377-87.
37. Bathaie SZ, Bolhassani A, Tamanoi F. Anticancer Effect and Molecular Targets of Saffron Carotenoids. Enzymes. 2014;36:57-86.
38. D'Alessandro AM, Mancini A, Lizzi AR, De Simone A, Marroccella CE, Gravina GL, et al. Crocus sativus stigma extract and its major constituent crocin possess significant antiproliferative properties against human prostate cancer. Nutr Cancer. 2013;65(6):930-42.
39. Mousavi B, Bathaie SZ, Fadai F, Ashtari Z, Ali Beigi N, Farhang S, et al. Safety evaluation of saffron stigma (Crocus sativus L.) aqueous extract and crocin in patients with schizophrenia. Avicenna J Phytomed. 2015;5(5):413-9.
40. Heidarzadeh H, Bathaie SZ, Abroun S, Mohagheghi MA. Evaluating the cytotoxic effect of crocin on MDA-MB-468 cell line based on apoptosis induction, ER stress, and autophagy markers. Pathobiology Research. 2018;20(4):37-51.
41. Debatin KM, Poncet D, Kroemer G. Chemotherapy: targeting the mitochondrial cell death pathway. Oncogene. 2002;21(57):8786-803.
42. Fulda S. Modulation of apoptosis by natural products for cancer therapy. Planta Med. 2010;76(11):1075-9.
43. Fesik SW. Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer. 2005;5(11):876-85.
44. Salih SM, Ringelstetter AK, Elsarrag MZ, Abbott DH, Roti EC. Dexrazoxane abrogates acute doxorubicin toxicity in marmoset ovary. Biol Reprod. 2015;92(3):73.
45. Bakshi HA, Hakkim FL, Sam S. Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation. Asian Pac J Cancer Prev. 2016;17(3):1499-506.
46. Mousavi SH, Tayarani NZ, Parsaee H. Protective effect of saffron extract and crocin on reactive oxygen species-mediated high glucose-induced toxicity in PC12 cells. Cell Mol Neurobiol. 2010;30(2):185-91.
47. Thomas E, Gopalakrishnan V, Somasagara RR, Choudhary B, Raghavan SC. Extract of Vernonia condensata, Inhibits Tumor Progression and Improves Survival of Tumor-allograft Bearing Mouse. Scientific reports. 2016;6:23255.
48. Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol. 2012;13(2):89-102.
| Files | ||
| Issue | Vol 11 No 1 (2019) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/bccr.v11i1.1646 | |
| Keywords | ||
| HER2 MTT Assay Flow Cytometry UPR Caspase-9 Anticancer | ||
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How to Cite
1.
Faridi N, Heidarzadeh H, Mohagheghi MA, Bathaie SZ. BT-474 Breast Cancer Cell Apoptosis Induced by Crocin, a Saffron Carotenoid. Basic Clin Cancer Res. 2019;11(1):5-15.
