CROCIN PROMOTES APOPTOSIS IN HUMAN EBV-TRANSFORMED B-LYMPHOCYTE VIA INTRINSIC PATHWAY CROCIN PROMOTES APOPTOSIS IN EBV-TRANSFORMED LCL
Main Article Content
Keywords
Crocin, CO 88BV59-1 EBV-Transformed Lymphoblastoid Cell Line (LCL), B-cell lymphoma, Apoptosis
Abstract
Background: As a major carotenoid in saffron, crocin demonstrates potent anti-cancer impacts. However, its anti-lymphoma effects remain vague, especially in the human EBV-associated B-cell lymphoproliferative disorders. This study examined crocin's apoptogenic potential and its underlying mechanism in CO 88BV59-1 cell line vs. normal human peripheral blood B cells.
Methods: CO 88BV59-1 cells were treated with crocin alone or in combination with vincristine for up to 72 h. The cell viability was examined using a resazurin assay. Flow cytometry using annexin V and propidium iodide labeling was performed to detect apoptotic cells. Also, the expression levels of genes and proteins involved in apoptosis (CASP3, CASP8, CASP9, P53, Bax, and Bcl-2) were respectively determined via real-time PCR and Western blot analysis.
Results: Crocin concentration-dependently reduced cell viability in CO 88BV59-1 cells with no significant toxicity toward normal B cells. Similar to vincristine, crocin significantly increased apoptosis in these cells during 72 h of incubation. Furthermore, the combination of crocin (80 μM) and vincristine (1 μM) enhanced apoptosis in CO 88BV59-1 cells. Therefore, this synergistic effect was detected in human EBV-transformed B-lymphocyte. CASP3, CASP9, P53, and Bax/Bcl-2 ratio expressions were significantly raised in CO 88BV59-1 cells, whereas CASP8 was unaltered. It was proposed that crocin promoted apoptosis in CO 88BV59-1 cells in a time- and concentration-dependent manner via the induction of the intrinsic pathway.
Conclusion: The results suggest that crocin may serve as a good alternative/coadjuvant to vincristine in EBV-associated B-cell lymphoproliferative disorders.
Keywords: Crocin, CO 88BV59-1 cells, EBV-associated B-cell lymphoproliferative disorders, apoptosis
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References
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[6] Khamisipour G, Jadidi-Niaragh F, Jahromi AS and Hojjat-Farsangi M. Mechanisms of tumor cell resistance to the current targeted-therapy agents. Tumor Biology 2016; 37: 10021-10039. https://doi.org/10.1007/s13277-016-5059-1
[7] Wegiera M, Smolarz HD, Jedruch M, Korczak M and Koproń K. Cytotoxic effect of some medicinal plants from Asteraceae family on J-45.01 leukemic cell line--pilot study. Acta poloniae pharmaceutica 2012; 69: 263-268. https://europepmc.org/article/med/22568040 PMID: 22568040
[8] Shu L, Cheung K-L, Khor TO, Chen C and Kong A-N. Phytochemicals: cancer chemoprevention and suppression of tumor onset and metastasis. Cancer and Metastasis Reviews 2010; 29: 483-502. https://doi.org/10.1007/s10555-010-9239-y
[9] Saklani A and Kutty SK. Plant-derived compounds in clinical trials. Drug discovery today 2008; 13: 161-171. https://doi.org/10.1016/j.drudis.2007.10.010
[10] Khorasanchi Z, Shafiee M, Kermanshahi F, Khazaei M, Ryzhikov M, Parizadeh MR, Kermanshahi B, Ferns GA, Avan A and Hassanian SM. Crocus sativus a natural food coloring and flavoring has potent anti-tumor properties. Phytomedicine 2018; 43: 21-27. https://doi.org/10.1016/j.phymed.2018.03.041
[11] Shakeri M, Tayer AH, Shakeri H, Jahromi AS, Moradzadeh M and Hojjat-Farsangi M. Toxicity of Saffron Extracts on Cancer and Normal Cells: A Review Article. Asian Pacific journal of cancer prevention: APJCP 2020; 21: 1867. doi: 10.31557/APJCP.2020.21.7.1867 PMID: 32711409
[12] Moradzadeh M, Tabarraei A, Ghorbani A, Hosseini A and Sadeghnia HR. Short‐term in vitro exposure to crocetin promotes apoptosis in human leukemic HL‐60 cells via intrinsic pathway. Acta Poloniae Pharm Drug Res 2018; 75: 445-451. https://scholar.google.com/scholar?hl=en&as_sdt=0%2C5&q=+Short%E2%80%90term+in+vitro+exposure+to+crocetin+promotes+apoptosis+in+human+leukemic+HL%E2%80%9060+cells+via+intrinsic+pathway.+Acta+Poloniae+Pharm+Drug+Res+2018%3B+75%3A+445-451.&btnG=
[13] Moradzadeh M, Ghorbani A, Erfanian S, Mohaddes ST, Rahimi H, Karimiani EG, Mashkani B, Chiang SC, El‐Khamisy SF and Tabarraei A. Study of the mechanisms of crocetin‐induced differentiation and apoptosis in human acute promyelocytic leukemia cells. Journal of Cellular Biochemistry 2019; 120: 1943-1957. https://doi.org/10.1002/jcb.27489
[14] Moore DK, Motaung B, du Plessis N, Shabangu AN, Loxton AG and Consortium S-I. Isolation of B-cells using Miltenyi MACS bead isolation kits. PloS one 2019; 14: e0213832. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0213832
[15] Mashkani B, Tanipour MH, Saadatmandzadeh M, Ashman LK and Griffith R. FMS-like tyrosine kinase 3 (FLT3) inhibitors: Molecular docking and experimental studies. European journal of pharmacology 2016; 776: 156-166. https://doi.org/10.1016/j.ejphar.2016.02.048
[16] Rangarajan P, Dharmalingam Subramaniam SP, Kwatra D, Palaniyandi K, Islam S, Harihar S, Ramalingam S, Gutheil W, Putty S and Pradhan R. Crocetinic acid inhibits hedgehog signaling to inhibit pancreatic cancer stem cells. Oncotarget 2015; 6: 27661. doi: 10.18632/oncotarget.4871
[17] Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic acids research 2001; 29: e45-e45. https://doi.org/10.1093/nar/29.9.e45
[18] Moradzadeh M, Tabarraei A, Sadeghnia HR, Ghorbani A, Mohamadkhani A, Erfanian S and Sahebkar A. Kaempferol increases apoptosis in human acute promyelocytic leukemia cells and inhibits multidrug resistance genes. Journal of Cellular Biochemistry 2018; 119: 2288-2297. https://doi.org/10.1002/jcb.26391
[19] Abts H, Emmerich M, Miltenyi S, Radbruch A and Tesch H. CD20 positive human B lymphocytes separated with the magnetic cell sorter (MACS) can be induced to proliferation and antibody secretion in vitro. Journal of Immunological Methods 1989; 125: 19-28. https://doi.org/10.1016/0022-1759(89)90073-2
[20] Marques-Piubelli ML, Salas YI, Pachas C, Becker-Hecker R, Vega F and Miranda RN. Epstein–Barr virus-associated B-cell lymphoproliferative disorders and lymphomas: a review. Pathology 2020; 52: 40-52. https://doi.org/10.1016/j.pathol.2019.09.006
[21] Gibson SE and Hsi ED. Epstein-Barr virus–positive B-cell lymphoma of the elderly at a United States tertiary medical center: an uncommon aggressive lymphoma with a nongerminal center B-cell phenotype. Human pathology 2009; 40: 653-661. https://doi.org/10.1016/j.humpath.2008.10.007
[22] Hoeller S, Tzankov A, Pileri SA, Went P and Dirnhofer S. Epstein-Barr virus–positive diffuse large B-cell lymphoma in elderly patients is rare in Western populations. Human pathology 2010; 41: 352-357. https://doi.org/10.1016/j.humpath.2009.07.024
[23] Rossi AP and Klein CL. Posttransplant malignancy. Surgical Clinics 2019; 99: 49-64. DOI:https://doi.org/10.1016/j.suc.2018.09.004
[24] Alavizadeh SH and Hosseinzadeh H. Bioactivity assessment and toxicity of crocin: a comprehensive review. Food and Chemical Toxicology 2014; 64: 65-80. https://doi.org/10.1016/j.fct.2013.11.016
[25] Moradzadeh M, Kalani MR and Avan A. The antileukemic effects of saffron (Crocus sativus L.) and its related molecular targets: A mini review. Journal of cellular biochemistry 2019; 120: 4732-4738. https://doi.org/10.1002/jcb.27525
[26] Li X, Jiang C and Zhu W. Crocin reduces the inflammation response in rheumatoid arthritis. Bioscience, Biotechnology, and Biochemistry 2017; 81: 891-898. https://doi.org/10.1080/09168451.2016.1263145
[27] Nam KN, Park Y-M, Jung H-J, Lee JY, Min BD, Park S-U, Jung W-S, Cho K-H, Park J-H and Kang I. Anti-inflammatory effects of crocin and crocetin in rat brain microglial cells. European journal of pharmacology 2010; 648: 110-116. https://doi.org/10.1016/j.ejphar.2010.09.003
[28] Tamaddonfard E, Farshid A-A, Eghdami K, Samadi F and Erfanparast A. Comparison of the effects of crocin, safranal and diclofenac on local inflammation and inflammatory pain responses induced by carrageenan in rats. Pharmacological Reports 2013; 65: 1272-1280. https://doi.org/10.1016/S1734-1140(13)71485-3
[29] Li K, Li Y, Ma Z and Zhao J. Crocin exerts anti-inflammatory and anti-catabolic effects on rat intervertebral discs by suppressing the activation of JNK. International journal of molecular medicine 2015; 36: 1291-1299. https://doi.org/10.3892/ijmm.2015.2359
[30] Tarantilis PA, Morjani H, POLISSIOU M and MANFAIT M. Inhibition of growth and induction of differentiation of promyelocytic leukemia (HL-60) by carotinoids from. Crocus sativus 1994; 1913-1918. https://scholar.google.com/scholar?hl=en&as_sdt=0%2C5&q=Tarantilis+PA%2C+Morjani+H%2C+POLISSIOU+M+and+MANFAIT+M.+Inhibition+of+growth+and+induction+of+differentiation+of+promyelocytic+leukemia+%28HL-60%29+by+carotinoids+from.+Crocus+sativus+1994%3B+1913-1918.&btnG=
[31] Morjani H, Tarantilis P, Polissiou M and Manfait M. Growth inhibition and induction of crythroid differentiation activity by crocin, dimethylcrocetine and β-carotene on K562 tumor cells. Anticancer Res 1990; 10: 1398-1406. https://scholar.google.com/scholar?hl=en&as_sdt=0%2C5&q=Morjani+H%2C+Tarantilis+P%2C+Polissiou+M+and+Manfait+M.+Growth+inhibition+and+induction+of+crythroid+differentiation+activity+by+crocin%2C+dimethylcrocetine+and+%CE%B2-carotene+on+K562+tumor+cells.+Anticancer+Res+1990%3B+10%3A+1398-1406.&btnG=
[32] Sun Y, Wang Z, Wang L, Wang L, Zang C and Sun L. The Effect and Mechanisms of Proliferative Inhibition of Crocin on Human Leukaemia Jurkat Cells. The West Indian medical journal 2015; 64: 473. doi: 10.7727/wimj.2016.053 PMID: 27398676
[33] Sun Y, Xu H-J, Zhao Y-X, Wang L-Z, Sun L-R, Wang Z and Sun X-F. Crocin exhibits antitumor effects on human leukemia HL-60 cells in vitro and in vivo. Evidence-Based Complementary and Alternative Medicine 2013; 2013: https://doi.org/10.1155/2013/690164
[34] Molnar J, Szabo D, Pusztai R, Mucsi I, Berek L, Ocsovszki I, Kawata E and Shoyama Y. Membrane associated antitumor effects of crocine-, ginsenoside-and cannabinoid derivates. Anticancer research 2000; 20: 861-867. https://europepmc.org/article/med/10810367 PMID: 10810367
[35] Bakshi HA, Sam S, Feroz A, Ravesh Z, Shah GA and Sharma M. Crocin from Kashmiri saffron (Crocus sativus) induces in vitro and in vivo xenograft growth inhibition of Dalton’s lymphoma (DLA) in mice. Asian Pac J Cancer Prev 2009; 10: 887-890. https://scholar.google.com/scholar?hl=en&as_sdt=0%2C5&q=Bakshi+HA%2C+Sam+S%2C+Feroz+A%2C+Ravesh+Z%2C+Shah+GA+and+Sharma+M.+Crocin+from+Kashmiri+saffron+%28Crocus+sativus%29+induces+in+vitro+and+in+vivo+xenograft+growth+inhibition+of+Dalton%E2%80%99s+lymphoma+%28DLA%29+in+mice.+Asian+Pac+J+Cancer+Prev+2009%3B+10%3A+887-890.&btnG=
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Jun 28, 2021
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Sotoodeh Jahromi, A. (2021) “CROCIN PROMOTES APOPTOSIS IN HUMAN EBV-TRANSFORMED B-LYMPHOCYTE VIA INTRINSIC PATHWAY : CROCIN PROMOTES APOPTOSIS IN EBV-TRANSFORMED LCL”, Mediterranean Journal of Hematology and Infectious Diseases, 13(1), p. e2021049. doi: 10.4084/MJHID.2021.049.
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