XAV939 promotes apoptosis in a neuroblastoma cell line via telomere shortening

Tian,Xiaohong; Hou,Weijian; Bai,Shuling; Fan,Jun; Tong,Hao; Bai,Yu

doi:10.3892/or.2014.3460

XAV939 promotes apoptosis in a neuroblastoma cell line via telomere shortening

Authors:
- Xiaohong Tian
- Weijian Hou
- Shuling Bai
- Jun Fan
- Hao Tong
- Yu Bai
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Affiliations: Department of Tissue Engineering, College of Basic Medical Sciences, China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Ophthalmology, The Second Hospital of Shijiazhuang, Shijiazhuang, Hebei 050051, P.R. China
Published online on: September 2, 2014 https://doi.org/10.3892/or.2014.3460
Pages: 1999-2006

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Abstract

Telomeres, telomerase and tankyrase (TNKS) have an extremely important and special association with human cell aging and cancer. Telomerase activity is abnormally high in cancer cells and is accompanied by the overexpression of tankyrase 1 (TNKS1). TNKS1 is a positive regulator of telomerase activation and telomere extension in the human body, indicating that TNKS1 may be a possible therapeutic target for cancer. XAV939 is a small-molecule inhibitor of TNKS1. The objective of the present study was to investigate the apoptotic effect of XAV939 on the neuroblastoma (NB) SH-SY5Y cell line, as well as the change in telomere length and telomerase activity and elucidate the mechanism from this perspective. In the present study, we initially treated SH-SY5Y cells with XAV939 and RNA interference (RNAi)-TNKS1, and subsequently chose the optimal sequence for RNAi-TNKS1. We then measured the telomere length using quantitative real-time polymerase chain reaction (qPCR) assay, detected the telomerase activity using the ELISA kit, observed apoptotic morphology by transmission electron microscopy, and detected the percentages of apoptotic cells using flow cytometry and Hoechst 33342 staining. We also determined the invasive ability by a cell invasion assay. The results showed that short hairpin RNA-2 (shRNA-2) was the optimal sequence for RNAi-TNKS1. Treatment with both XAV939 and RNAi-TNKS1 shortened the telomere length, promoted apoptosis and reduced the invasive ability of the SH-SY5Y cells, yet had no effect on telomerase activity. XAV939 promoted apoptosis and reduced the invasiveness of SH-SY5Y cells dependent on telomere shortening, and further research should be conducted to clarify the exact mechanisms. This research may contribute to the cure of malignant NB using multi-targeted therapy with small-molecule agents.

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1	Ramirez R, Carracedo J, Jiménez R, et al: Massive telomere loss is an early event of DNA damage-induced apoptosis. J Biol Chem. 278:836–842. 2003. View Article : Google Scholar : PubMed/NCBI
2	Lyons RJ, Deane R, Lynch DK, Ye ZS, Sanderson GM, Eyre HJ, Sutherland GR and Daly RJ: Identification of a novel human tankyrase through its interaction with the adaptor protein Grbl4. J Biol Chem. 276:17172–17180. 2001. View Article : Google Scholar : PubMed/NCBI
3	De Rycker M, Venkatesan RN, Wei C and Price CM: Vertebrate tankyrase domain structure and sterile α motif (SAM)-mediated multimerization. Biochem J. 372:87–96. 2003.
4	Seimiya H, Muramatsu Y, Smith S and Tsuruo T: Functional subdomain in the ankyrin domain of tankyrase 1 required for poly(ADP-ribosyl)ation of TRF1 and telomere elongation. Mol Cell Biol. 24:1944–1955. 2004. View Article : Google Scholar : PubMed/NCBI
5	De Rycker M and Price CM: Tankyrase polymerization is controlled by its sterile α motif and poly(ADP-ribose) polymerase domains. Mol Cell Biol. 24:9802–9812. 2004.PubMed/NCBI
6	Kaminker PG, Kim SH, Taylor RD, Zebarjadian Y, Funk WD, Morin GB, Yaswen P and Campisi J: TANK2, a new TRF1-associated poly(ADP-ribose) polymerase, causes rapid induction of cell death upon overexpression. J Biol Chem. 276:35891–35899. 2001. View Article : Google Scholar : PubMed/NCBI
7	Seimiya H: The telomeric PARP, tankyrases, as targets for cancer therapy. Br J Cancer. 94:341–345. 2006. View Article : Google Scholar : PubMed/NCBI
8	Smith S, Giriat I, Schmitt A and de Lange T: Tankyrase, a poly(ADP-ribose) polymerase at human telomeres. Science. 282:1484–1487. 1998. View Article : Google Scholar : PubMed/NCBI
9	Hsiao SJ and Smith S: Sister telomeres rendered dysfunctional by persistent cohesion are fused by NHEJ. J Cell Biol. 184:515–526. 2009. View Article : Google Scholar : PubMed/NCBI
10	Papeo G, Forte B, Orsini P, Perrera C, Posteri H, Scolaro A and Montagnoli A: Poly(ADP-ribose) polymerase inhibition in cancer therapy: are we close to maturity? Expert Opin Ther Pat. 19:1377–1400. 2009. View Article : Google Scholar : PubMed/NCBI
11	Muramatsu Y, Ohishi T, Sakamoto M, Tsuruo T and Seimiya H: Cross-species difference in telomeric function of tankyrase 1. Cancer Sci. 98:850–857. 2007. View Article : Google Scholar : PubMed/NCBI
12	Lu R, Pal J, Buon L, et al: Targeting homologous recombination and telomerase in Barrett’s adenocarcinoma: impact on telomere maintenance, genomic instability and tumor growth. Oncogene. 33:1495–1505. 2014.PubMed/NCBI
13	Seimiya H, Muramatsu Y, Ohishi T and Tsuruo T: Tankyrase 1 as a target for telomere-directed molecular cancer therapeutics. Cancer Cell. 7:25–37. 2005. View Article : Google Scholar : PubMed/NCBI
14	Tian XH, Hou WJ, Fang Y, Fan J, Tong H, Bai SL, Chen Q, Xu H and Li Y: XAV939, a tankyrase 1 inhibitior, promotes cell apoptosis in neuroblastoma cell lines by inhibiting Wnt/β-catenin signaling pathway. J Exp Clin Cancer Res. 32:1002013.PubMed/NCBI
15	Huang SM, Mishina YM, Liu S, et al: Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling. Nature. 461:614–620. 2009. View Article : Google Scholar : PubMed/NCBI
16	Dregalla RC, Zhou J, Idate RR, Battaglia CL, Liber HL and Bailey SM: Regulatory roles of tankyrase 1 at telomeres and in DNA repair: suppression of T-SCE and stabilization of DNA-PKcs. Aging. 2:691–708. 2010.PubMed/NCBI
17	Sun J, Huang H and Zhu YY: Study on the expression of tankyrase in malignant hematopoietic cells and its relation with telomerase activity. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 12:11–15. 2004.(In Chinese).
18	Cawthon RM: Telomere measurement by quantitative PCR. Nucleic Acids Res. 30:e472002. View Article : Google Scholar : PubMed/NCBI
19	Cawthon RM: Telomere length measurement by a novel monochrome multiplex quantitative PCR method. Nucleic Acids Res. 37:e212009. View Article : Google Scholar : PubMed/NCBI
20	Gelmini S, Poggesi M, Distante V, Bianchi S, Simi L, Luconi M, Raggi CC, Cataliotti L, Pazzagli M and Orlando C: Tankyrase, a positive regulator of telomere elongation, is overexpressed in human breast cancer. Cancer Lett. 216:81–87. 2004. View Article : Google Scholar : PubMed/NCBI
21	Gelmini S, Quattrone S, Malentacchi F, Villari D, Travaglini F, Giannarini G, Della Melina A, Pazzagli M, Nicita G, Selli C and Orlando C: Tankyrase-1 mRNA expression in bladder cancer and paired urine sediment: preliminary experience. Clin Chem Lab Med. 45:862–866. 2007. View Article : Google Scholar : PubMed/NCBI
22	Poonepalli A, Banerjee B, Ramnarayanan K, Palanisamy N, Putti TC and Hande MP: Telomere-mediated genomic instability and the clinico-pathological parameters in breast cancer. Genes Chromosomes Cancer. 47:1098–1109. 2008. View Article : Google Scholar : PubMed/NCBI
23	Gelmini S, Poggesi M, Pinzani P, Mannurita SC, Cianchi F, Valanzano R and Orlando C: Distribution of Tankyrase-1 mRNA expression in colon cancer and its prospective correlation with progression stage. Oncol Rep. 16:1261–1266. 2006.PubMed/NCBI
24	Shay JW and Bacchetti S: A survey of telomerase activity in human cancer. Eur J Cancer. 33:787–791. 1997. View Article : Google Scholar : PubMed/NCBI
25	Andrews LG and Tollefsbol TO: Methods of telomerase inhibition. Methods Mol Biol. 405:1–8. 2007. View Article : Google Scholar : PubMed/NCBI
26	Aubert G and Lansdorp PM: Telomeres and aging. Physiol Rev. 88:557–579. 2008. View Article : Google Scholar
27	Baerlocher GM, Vulto I, de Jong G and Lansdorp PM: Flow cytometry and FISH to measure the average length of telomeres (flow FISH). Nat Protoc. 1:2365–2376. 2006. View Article : Google Scholar : PubMed/NCBI
28	Poon SS and Lansdorp PM: Measurements of telomere length on individual chromosomes by image cytometry. Methods Cell Biol. 64:69–96. 2001. View Article : Google Scholar : PubMed/NCBI
29	Zhang H, Yang MH, Zhao JJ, Chen L, Yu ST, Tang XD, Fang DC and Yang SM: Inhibition of tankyrase 1 in human gastric cancer cells enhances telomere shortening by telomerase inhibitors. Oncol Rep. 24:1059–1065. 2010.PubMed/NCBI
30	Cerone MA, Burgess DJ, Naceur-Lombardelli C, Lord CJ and Ashworth A: High-throughput RNAi screening reveals novel regulators of telomerase. Cancer Res. 71:3328–3340. 2011. View Article : Google Scholar : PubMed/NCBI
31	Bilsland AE, Hoare S, Stevenson K, et al: Dynamic telomerase gene suppression via network effects of GSK3 inhibition. PLoS One. 4:e64592009. View Article : Google Scholar : PubMed/NCBI
32	Ohishi T, Tsuruo T and Seimiya H: Evaluation of tankyrase inhibition in whole cells. Methods Mol Biol. 405:133–146. 2007. View Article : Google Scholar : PubMed/NCBI
33	Xu D, Li H and Liu JP: Inhibition of telomerase by targeting MAP kinase signaling. Methods Mol Biol. 405:147–165. 2007. View Article : Google Scholar : PubMed/NCBI
34	Lau T, Chan E, Callow M, Waaler J, Boggs J, Blake RA, Magnuson S, Sambrone A, Schutten M, Firestein R, Machon O, Korinek V, Choo E, Diaz D, Merchant M, Polakis P, Holsworth DD, Krauss S and Costa M: A novel tankyrase small-molecule inhibitor suppresses APC mutation-driven colorectal tumor growth. Cancer Res. 73:3132–3144. 2013. View Article : Google Scholar : PubMed/NCBI
35	Kim MK and Smith S: Persistent telomere cohesion triggers a prolonged anaphase. Mol Biol Cell. 25:30–40. 2014. View Article : Google Scholar : PubMed/NCBI
36	Dynek JN and Smith S: Resolution of sister telomere association is required for progression through mitosis. Science. 304:97–100. 2004. View Article : Google Scholar : PubMed/NCBI
37	Chang P, Coughlin M and Mitchison TJ: Tankyrase-1 polymerization of poly(ADP-ribose) is required for spindle structure and function. Nat Cell Biol. 7:1133–1139. 2005. View Article : Google Scholar : PubMed/NCBI