Effects of annexin A1 on apoptosis and cell cycle arrest in human leukemic cell lines

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Recent studies suggest that annexin A1 (ANXA1) promotes apoptosis in cancerous cells. This study aims to investigate the effects of ANXA1 on apoptosis and cell cycle arrest in K562, Jurkat and U937 cells and peripheral blood mononu-clear cells (PBMC). Cells were treated with ANXA1 and cyclophosphamide prior to flow cytometry analysis for apoptosis and cell cycle arrest induction. At 2.5µM, ANXA1 induced significant apoptosis in K562 (p ≤ 0.001) and U937 (p ≤ 0.05) cells, with EC50 values of 3.6 and 3.8 µM, respectively. In Jurkat cells, induction was not significant (EC50, 17.0 µM). No significant apoptosis induction was observed in PBMC. ANXA1 caused cycle arrest in the G0/G1 phase in K562 and U937 cells with p ≤ 0.001 for both, and (p ≤ 0.01) for Jurkat cells. ANXA1 induced apoptosis and cycle arrest in the G0/G1 phase in K562 and U937 cells, causing only cell cycle arrest in Jurkat cells.

1. W. J. Liu, T. Zhang, Q. L. Guo, C. Y. Liu and Y. Q. Bai, Effect of ATRA on the expression of HOXA5 gene in K562 cells and its relationship with cell cycle and apoptosis, Mol. Med. Rep. 13 (2016) 4221–4228; 10.3892/mmr.2016.5086

2. K. Yata, Y. Sadahira, T. Otsuki, H. Sakaguchi, Y. Isozaki and M. Uno, Cell cycle analysis and expression of cell cycle regulator genes in myeloma cells overexpressing cyclin D1, Br. J. Haematol. 114 (2001) 591–599.

3. B. T. Gjertsen and H. Wiig, Investigation of therapy resistance mechanisms in myeloid leukemia by protein profiling of bone marrow extracellular fluid, Expert Rev. Proteomics 9 (2012) 595–598; https://doi.org/10.1586/epr.12.55

4. W. Ni, W. Qian and X. Tong, Cryptotanshinone induces apoptosis of HL-60 cells via mitochondrial pathway, Trop. J. Pharm. Res. 13 (2014) 545–551.

5. S. Agrawal, M. Unterberg, S. Koschmieder, U. zur Stadt, U. Brunnberg, W. Verbeek, T. Büchner, W. E. Berdel, H. Serve and C. Müller-Tidow, DNA methylation of tumor suppressor genes in clinical remission predicts the relapse risk in acute myeloid leukemia, Cancer Res. 67 (2007) 1370–1377; https://doi.org/10.1158/0008-5472.CAN-06-1681

6. Q. Y. Lu, Y. S. Jin, Z. F. Zhang, A. D. Le, D. Heber, F. P. Li, S. M. Dubinett and J. Y. Rao, Green tea induces annexin-I expression in human lung adenocarcinoma A549 cells: involvement of annex-in-I in actin remodeling, Lab. Investig. 87 (2007) 456–465; http://doi.org/10.1038/labinvest.3700534

7. S. H. Xia, L. P. Hu, H. Hu, W. T. Ying, X. Xu, Y. Cai, Y. L. Han, B. S. Chen, F. Wei, X. H. Qian, Y. Y. Cai, Y. Shen, M. Wu and M. R. Wang, Three isoforms of annexin I are preferentially expressed in normal esophageal epithelia but down-regulated in esophageal squamous cell carcinomas, Onco-gene 21 (2002) 6641–6648

8. R. Luthra, R. R. Singh, M. G. Luthra, Y. X. Li, C. Hannah, A. M. Romans, B. A. Barkoh, S. S. Chen, J. Enros, D. M. Maru, R. R. Boraddus, A. Rashid and C. T. Albarracin, MicroRNA-196a targets annexin A1: a microRNA-mediated mechanism of annexin A1 downregulation in cancers, Oncogene 27 (2008) 6667–6678; https://doi.org/10.1038/onc.2008.256

9. R. Duncan, B. Carpenter, L. C. Main, C. Telfer and G. I. Murray, Characterisation and protein expression profiling of annexins in colorectal cancer, Br. J. Cancer. 98 (2008) 426–433; https://doi.org/10.1038/sj.bjc.6604128

10. R. Belvedere, V. Bizzarro, G. Forte, F. D. Piaz, L. Parente and A. Petrella, Annexin A1 contributes to pancreatic cancer cell phenotype, behaviour and metastatic potential independently of formyl peptide receptor pathway, Sci. Reports 6 (2016) 1–14.

11. W. Ahmad, E. Kumolosasi, I. Jantan, M. Jasamai and E. Salim, Modulatory effect of phytoestrogens and curcumin on induction of annexin 1 in human peripheral blood mononuclear cells and their inhibitory effect on secretory phospholipase A2, Trop. J. Pharm. Res. 13 (2014) 171–177; https://doi.org/10.4314/tjpr.v13i2.1

12. L. Parente and E. Solito, Annexin 1: More than an anti-phospholipase protein, Inflamm. Res. 53 (2004) 125–132; https://doi.org/10.1007/s00011-003-1235-z

13. L. H. K. Lim and S. Pervaiz, Annexin 1 : the new face of an old molecule, FASEB J. 24 (2007) 968–975; https://doi.org/10.1096/fj.06-7464rev

14. C. Guo, S. Liu and M-Z. Sun, Potential role of Anxa1 in cancer, Futur. Oncol. 9 (2013) 1773–1793; https://doi.org/10.2217/fon.13.114

15. S. Canaider, E. Solito, C. de Coupade, R. J, Flower, F. Russo-Marie, N. J. Goulding and M. Peretti, Increased apoptosis in U937 cells over-expressing lipocortin 1 (Annexin I), Life Sci. 66 (2000) PL265-PL270

16. E. Solito E, C. De Coupade, S. Canaider, N. J. Goulding and M. Perretti, Transfection of annexin 1 in monocytic cells produces a high degree of spontaneous and stimulated apoptosis associated with caspase-3 activation, Br. J. Pharmacol. 13 (2001) 217–228; https://doi.org/10.1038/sj.bjp.0704054

17. F. Hirata, Molecular targets, cancer chemoprevention, and dietary phytochemicals: Nuclear annexin A1 as a promising new molecular target of cancer chemoprevention, Austin J. Pharmacol. Ther. 2 (2014) 2–3.

18. A. Petrella, C. W. D’Acunto, M. Rodriquez, M. Festa, A, Tosco, I. Bruno, S. Terracciano, M. Taddei, L. G. Paloma and L. Parente, Effects of FR235222, a novel HDAC inhibitor, in proliferation and apoptosis of human leukaemia cell lines: Role of annexin, Eur. J. Cancer. 44 (2008) 740–749; https://doi.org/10.1016/j.ejca.2008.01.023

19. C. L. Rackham, A. E. Vargas, R. G. Hawkes, S. Amisten, S. J. Persaud, A. L. Austin, A. J. King and P. M. Jones, Annexin A1 is a key modulator of mesenchymal stromal cell-mediated improvements in islet function, Diabetes 65 (2016) 129–139; https://doi.org/10.2337/db15-0990

20. G. S. D. Purvis, F. Chiazza, J. Chen, R. Azevedo-Loiola, K. Martin, D. H. M. Kusters, C. C. Reutelingsperger, N. Fountoulakis, L. Gnudi, M. M. Yaqoob, M. Collino, C. Thiemermann and E. Solito, Annexin A1 attenuates microvascular complications through restoration of Akt signalling in a murine model of type 1 diabetes, Diabetologia 61 (2018) 482–495; https://doi.org/10.1007/s00125-017-4469-y

21. E. Cristante, S. McArthur, C. Mauro, E. Maggioli, I. A. Romero, M. Wylezinska-Arridge, P. O. Couraud, J. lopez-Tremoleda, H. C. Christian, B. B. Weksler, A. Malaspina and E. Solito, Identification of an essential endogenous regulator of blood-brain barrier integrity, and its pathological and therapeutic implications, Proc. Natl. Acad. Sci. USA 110 (2012) 832–841; https://doi.org/10.1073/pnas.1209362110

22. M. Ries, R. Loiola, U. N. Shah, S. M. Gentleman, E. Solito and M. Sastre, The anti-inflammatory Annexin A1 induces the clearance and degradation of the amyloid-β peptide, J. Neuroinflammation (2016) 1–15; https://doi.org/10.1186/s12974-016-0692-6

23. M. F. Maioral, P. C. Gaspar, G. R. Rosa Souza, A. Mascarello, L. D. Chiaradia, M. A. Licínio, A. C. Moraes, R. A. Yunes, R. J. Nunes and M. C. Santos-Silva, Apoptotic events induced by synthetic naphthylchalcones in human acute leukemia cell lines, Biochimie 95 (2013) 866–874; https://doi.org/10.1016/j.biochi.2012.12.001

24. M. Luczak, M. Kaźmierczak, L. Handschuh, K. Lewandowski, M. Komarnicki and M. Figlerowicz, Comparative proteome analysis of acute myeloid leukemia with and without maturation, J. Proteomics 75 (2012) 5734–5748; https://doi.org/10.1016/j.jprot.2012.07.030

25. R. Debret, H. El Btaouri, L. Duca, I. Rahman, S. Radke, B. Haye, J. M. Sallenave and F. Antonicelli, Annexin A1 processing is associated with caspase-dependent apoptosis in BZR cells, FEBS Lett. 546 (2003) 195–202.

26. J. P. Vago, C. R. Nogueira, L. P. Tavares, F. M. Soriani, F. Lopes, R. C. Russo, V. Pinho, M. M. Teixeira and L. P. Sousa, Annexin A1 modulates natural and glucocorticoid-induced resolution of inflammation by enhancing neutrophil apoptosis, J. Leukoc. Biol. 92 (2012) 249–258; http://doi.org/10.1189/jlb.0112008

27. G. Li, S. He, L. Chang, H. Lu, H. Zhang, H. Zhang and J. Chiu, GADD45α and annexin A1 are involved in the apoptosis of HL-60 induced by resveratrol, Phytomedicine 18 (2011) 704–709; https://doi.org/10.1016/j.phymed.2010.11.015

28. R. N. Winter, A. Kramer, A. Borkowski and N. Kyprianou, Loss of caspase-1 and caspase-3 protein expression in human prostate cancer, Cancer Res. 61 (2001) 1227–1232.

29. T. C. Reddy, D. B. Reddy, A. Aparna, K. M. Arunasree, G. Gupta, C. Achari, G. V. Reddy, V. Lakshmipathi, A. Subramanyam and P. Reddanna, Anti-leukemic effects of gallic acid on human leukemia K562 cells: downregulation of COX-2, inhibition of BCR/ABL kinase and NF-ĸB inactivation, Toxicol. In Vitro 26 (2012) 396–405; https://doi.org/10.1016/j.tiv.2011.12.018

30. S. H. Ferreira, F. Q. Cunha, B. B. Lorenzetti, M. A. Michelin, M. Perretti, R. J. Flower and S. Poole, Role of lipocortin-1 in the anti-hyperalgesic actions of dexamethasone, Br. J. Pharmacol. 121 (1997) 883–888; https://doi.org/10.1038/sj.bjp.0701211

31. Y. Nakanishi, R. Kamijo, K. Takizawa, M. Hatori and M. Nagumo, Inhibitors of cyclooxygenase-2 (COX-2) suppressed the proliferation and differentiation of human leukaemia cell lines, Eur. J. Cancer. 37 (2001) 1570–1578.

32. A. M. Saleh, A. Aljada, M. M. El-Abadelah, M. O. Taha, S. S. Sabri, J. A. Zahra and M. A. Aziz, The anticancer activity of the substituted pyridone-annelated isoindigo (5’-Cl) involves G0/G1 cell cycle arrest and inactivation of CDKs in the promyelocytic leukemia cell line HL-60, Cell Physiol. Biochem. 35 (2015) 1943–1957; https://doi.org/10.1159/000374003

33. Y. Nagahara, Y. Matsuoka, K. Saito, M. Ikekita, S. Higuchi and T. Shinomiya, Coordinate involvement of cell cycle arrest and apoptosis strengthen the effect of FTY720, Jap. J. Cancer Res. 92 (2001) 680–687.

34. R. A. Weinberg, p53 and Apoptosis: Master Guardian and Executioner, in The Biology of Cancer (Ed. R. A. Weinberg), 2nd ed, Garland Science, New York 2014. pp. 331–390.

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