1. bookVolume 6 (2012): Issue 3 (June 2012)
Journal Details
License
Format
Journal
First Published
01 Jun 2007
Publication timeframe
6 times per year
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English
Copyright
© 2020 Sciendo
Journal Details
License
Format
Journal
First Published
01 Jun 2007
Publication timeframe
6 times per year
Languages
English
Copyright
© 2020 Sciendo
Abstract

Background: The alternative splicing of Bcl-x generates the proapoptotic Bcl-xs protein and the antiapoptotic variant Bcl-xl. Previous studies have demonstrated that some chemotherapeutic agents such as emetine, staurosporine, and epigallocatechin gallate (EGCG) in combination with ibuprofen significantly altered the ratio of the Bcl-x variants Bcl-xs/Bcl-xl in various cell lines, suggesting Bcl-x splicing might be affected by the exogenous stimuli.

Objective: We investigated the regulative role of imatinib in the alternative pre-mRNA splicing of Bcl-x in K562 cells and the related mechanism.

Methods: Cell proliferation was measured using WST assay kit. Cell apoptosis was assayed using an Annexin V-FITC Apoptosis Detection Kit. RT-PCR and western blot assay was used to analyze the mRNA and protein level of alternative splicing of exon 2 in the Bcl-x gene respectively.

Results: Imatinib regulated the alternative splicing in the Bcl-x gene in the K562 cells. In addition, we found that hydroxyurea, another agent for the therapy of CML, could enhance the effect of imatinib on the ratio of the Bcl-xl/Bcl-xs. Moreover, the induction of alternative splicing was correlated with protein phosphatase 1 (PP1). Alternatively, pretreatment with calyculin efficiently blocked imatinib-induced alternative splcing in the K562 cells compared with okadaic acid, which showed an important role of PP1 in regulating imatinib-induced splicing.

Conclusion: Imatinib regulates the alternative splicing of Bcl-x in K562 cells, which may be associated with the activation of PP1.

Keywords

1. Wang ET, Sandberg R, Luo S, Khrebtukova I, Zhang L, Mayr C, et al. Alternative isoform regulation in human tissue transcriptomes. Nature. 2008; 456:470-6.Search in Google Scholar

2. Srebrow A, Kornblihtt AR. The connection between splicing and cancer. J Cell Sci 2006;119:2635-41.Search in Google Scholar

3. Cooper TA, Wan L, Dreyfuss G. RNA and disease. Cell. 2009; 136:777-93.Search in Google Scholar

4. Akgul C, Moulding DA, Edwards SW. Alternative splicing of Bcl-2-related genes: functional consequences and potential therapeutic applications. Cell Mol Life Sci. 2004; 61:2189-99.Search in Google Scholar

5. Shkreta L, Froehlich U, Paquet ER, Toutant J, Elela SA, Chabot B.Anticancer drugs affect the alternative splicing of Bcl-x and other human apoptotic genes. Mol Cancer Ther. 2008;7:1398-409.10.1158/1535-7163.MCT-08-0192Open DOISearch in Google Scholar

6. Druker BJ, Tamura S, Buchdunger E, Ohno S, Segal GM, Fanning S, et al. Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nat Med. 1996; 2:561-66.10.1038/nm0596-561Open DOISearch in Google Scholar

7. Hughes T, Deininger M, Hochhaus A, Branford S, Radich J, Kaeda J, et al. Monitoring CML patients responding to treatment with tyrosine kinase inhibitors: review and recommendations for harmonizing current methodology for detecting BCR-ABL transcripts and kinase domain mutations and for expressing results. Blood. 2006; 108:28-37.Search in Google Scholar

8. Druker BJ, Guilhot F, O’Brien SG, Gathmann I, Kantarjian H, Gattermann N, et al. Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N Engl J Med. 2006; 355:2408-17.Search in Google Scholar

9. Carroll M, Ohno-Jones S, Tamura S, Buchdunger E, Zimmermann J, Lydon NB, et al. CGP 57148, a tyrosine kinase inhibitor, inhibits the growth of cells expressing BCR-ABL, TEL-ABL, and TEL-PDGFR fusion proteins. Blood. 1997; 90:4947-52.Search in Google Scholar

10. Wang AH, Wang YY, Yao Y, Xu ZZ, Zhou L, Wang L, et al. Summary of 615 patients of chronic myeloid leukemia in Shanghai from 2001 to 2006. J Exp Clin Cancer Res. 2010;29:20.Search in Google Scholar

11. de Lavallade H, Apperley JF, Khorashad JS, Milojkovic D, Reid AG, Bua M, et al. Imatinib for newly diagnosed patients with chronic myeloid leukemia: incidence of sustained responses in an intention-totreat analysis. J Clin Oncol. 2008; 26:3358-63.10.1200/JCO.2007.15.8154Open DOISearch in Google Scholar

12. Kano Y, Akutsu M, Tsunoda S, Mano H, Sato Y, Honma Y, et al. In vitro cytotoxic effects of a tyrosine kinase inhibitor STI571 in combination with commonly used antileukemic agents. Blood. 2001; 97:1999-2007.Search in Google Scholar

13. Gerl R, Vaux DL. Apoptosis in the development and treatment of cancer. Carcinogenesis. 2005; 26:263-70.10.1093/carcin/bgh283Open DOISearch in Google Scholar

14. Pajares MJ, Ezponda T, Catena R, Calvo A, Pio R, Montuenga LM. Alternative splicing: an emerging topic in molecular and clinical oncology. Lancet Oncol. 2007; 8:349-57.10.1016/S1470-2045(07)70104-3Open DOISearch in Google Scholar

15. Revil T, Toutant J, Shkreta L, Garneau D, Cloutier P, Chabot B: Protein kinase C-dependent control of Bcl-x alternative splicing. Mol Cell Biol. 2007; 27: 8431-41.10.1128/MCB.00565-07Open DOISearch in Google Scholar

16. Kim MH. Protein phosphatase 1 activation and alternative splicing of Bcl-X and Mcl-1 by EGCG + ibuprofen. J Cell Biochem. 2008; 104:1491-9.Search in Google Scholar

17. Chalfant CE, Rathman K, Pinkerman RL, Wood RE, Obeid LM, Ogretmen B, et al. De novo ceramide regulates the alternative splicing of caspase 9 and Bcl-x in A549 lung adenocarcinoma cells. Dependence on protein phosphatase-1. J Biol Chem. 2002; 277: 12587-95.Search in Google Scholar

18. Du K, Peng Y, Greenbaum LE, Haber BA, Taub R. HRS/SRp40-mediated inclusion of the fibronectin EIIIB exon, a possible cause of increased EIIIB expression in proliferating liver. Mol Cell Biol. 1997; 17:4096-104.10.1128/MCB.17.7.4096Open DOISearch in Google Scholar

19. Patel NA, Chalfant CE, Watson JE, Wyatt JR, Dean NM, Eichler DC, et al. Insulin regulates alternative splicing of protein kinase C beta II through a phosphatidylinositol 3-kinase-dependent pathway involving the nuclear serine/arginine-rich splicing factor, SRp40, in skeletal muscle cells. J Biol Chem. 2001; 276:22648-54.Search in Google Scholar

20. Ghosh N, Patel N, Jiang K, Watson JE, Cheng J, Chalfant CE, et al. Ceramide-activated protein phosphatase involvement in insulin resistance via Akt, serine/arginine-rich protein 40, and ribonucleic acid splicing in L6 skeletal muscle cells. Endocrinology. 2007; 148:1359-66.Search in Google Scholar

21. Blaustein M, Pelisch F, Coso OA, Bissell MJ, Kornblihtt AR, Srebrow A. Mammary epithelialmesenchymal interaction regulates fibronectin alternative splicing via phosphatidylinositol 3-kinase. J Biol Chem. 2004; 279:21029-37.Search in Google Scholar

22. Pelisch F, Blaustein M, Kornblihtt AR, Srebrow A. Cross-talk between signaling pathways regulates alternative splicing: a novel role for JNK. J Biol Chem. 2005; 280:25461-9.Search in Google Scholar

23. Horita M, Andreu EJ, Benito A, Arbona C, Sanz C, Benet I, et al. Blockade of the Bcr-Abl kinase activity induces apoptosis of chronic myelogenous leukemia cells by suppressing signal transducer and activator of transcription 5-dependent expression of Bcl-xL. J Exp Med. 2000; 191:977-84.Search in Google Scholar

24. Dresemann G. Imatinib and hydroxyurea in pretreated progressive glioblastoma multiforme: a patient series. Ann Oncol. 2005; 16:1702-8.10.1093/annonc/mdi317Open DOISearch in Google Scholar

25. Reardon DA, Egorin MJ, Quinn JA, Rich JN, Gururangan S, Vredenburgh JJ, et al. Phase II study of imatinib mesylate plus hydroxyurea in adults with recurrent glioblastoma multiforme. J Clin Oncol. 2005; 23:9359-68.10.1200/JCO.2005.03.2185Open DOISearch in Google Scholar

26. Skorta I, Oren M, Markwardt C, Gutekunst M, Aulitzky WE, van der Kuip H. Imatinib mesylate induces cisplatin hypersensitivity in Bcr-Abl+ cells by differential modulation of p53 transcriptional and proapoptotic activity. Cancer Res. 2009; 69:9337-45.10.1158/0008-5472.CAN-09-0548Open DOISearch in Google Scholar

27. Kuret J, Bell H, Cohen P. Identification of high levels of protein phosphatase-1 in rat liver nuclei. FEBS Lett. 1986; 203:197-202.Search in Google Scholar

28. Misteli T, Spector DL. Serine/threonine phosphatase 1 modulates the subnuclear distribution of pre-mRNA splicing factors. Mol Biol Cell. 1996; 7:1559-72.10.1091/mbc.7.10.1559Open DOISearch in Google Scholar

29. Chalfant CE, Ogretmen B, Galadari S, Kroesen BJ, Pettus BJ, Hannun YA. FAS activation induces dephosphorylation of SR proteins; dependence on the de novo generation of ceramide and activation of protein phosphatase 1. J Biol Chem. 2001; 276: 44848-55.Search in Google Scholar

30. Mermoud JE, Cohen PT, Lamond AI. Regulation of mammalian spliceosome assembly by a protein phosphorylation mechanism. EMBO J. 1994; 13: 5679-88.Search in Google Scholar

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