Human phosphatidylethanolamine-binding protein 4(hPEBP4) is a novel anti-apoptosis molecule associated with the resistance of tumors to apoptotic agents. Here we sought to investigate the role of hPEBP4 in the radioresistance of rectal cancer. Immunohistochemistry analysis showed hPEBP4 was expressed in 27/33 of rectal cancer specimens, but only in 2/33 of neighboring normal mucosa. Silencing the expression of hPEBP4 with siRNA significantly reduced the clonogenic survival and enhanced the apoptosis of rectal cancer cells on irradiation. Instead, forced overexpression of hPEBP4 promoted its survival and decreased the apoptosis. Western blot showed hPEBP4 could increase the radiation-induced Akt activation, for which reactive oxygen specimen(ROS) was required. The radioresistance effect of hPEBP4 was reversed after given LY-294002 to inhibit Akt activation or antioxidant to abolish the ROS production. We also confirmed that effect of hPEBP4 in vivo with nude mice. Thus we concluded that hPEBP4, specifically expressed in rectal cancer cells, is associated with radioresistance of rectal cancer, implying that modulation of hPEBP4 may have important therapeutic implications in radiotherapy of rectal cancer.
References
[1]
Siegel R, DeSantis C, Virgo K, Stein K, Mariotto A, et al. (2012) Cancer Treatment and Survivorship Statistics. CA Cancer J Clin 62: 220–241. doi: 10.3322/caac.21149
[2]
Sauer R, Becker H, Hohenberger W, R？del C, Wittekind C, et al. (2004) Preoperative versus postoperative chemoradiotherapy for rectal cancer. N Engl J Med 351: 1731–1740. doi: 10.1056/nejmoa040694
[3]
Krishnan S, Janjan NA, Skibber JM, Rodriguez-Bigas MA, Wolff RA, et al. (2006) Phase II study of capecitabine (Xeloda) and concomitant boost radiotherapy in patients with locally advanced rectal cancer. Int J Radiat Oncol Biol Phys 66: 762–771. doi: 10.1016/j.ijrobp.2006.05.063
[4]
Wang X, Li N, Liu B, Sun H, Chen T, et al. (2004) A novel human phosphatidylethanolamine-binding protein resists tumor necrosis factor alpha-induced apoptosis by inhibiting mitogen-activated protein kinase pathway activation and phosphatidylethanolamine externalization. J Biol Chem 279: 45855–45864. doi: 10.1074/jbc.m405147200
[5]
Wang X, Li N, Li H, Liu B, Qiu J, et al. (2005) Silencing of human phosphatidylethanolamine-binding protein 4 sensitizes breast cancer cells to TNF-α-induced apoptosis and cell growth arrest. Clin Cancer Res 11: 7545–7553. doi: 10.1158/1078-0432.ccr-05-0879
[6]
Li H, Wang X, Li N, Qiu J, Zhang Y, et al. (2007) hPEBP4 resists TRAIL-induced apoptosis of human prostate cancer cells by activating Akt and deactivating ERK1/2 pathways. J Biol Chem 282: 4943–4950. doi: 10.1074/jbc.m609494200
[7]
Li P, Wang X, Li N, Kong H, Guo Z, et al. (2006) Anti-apoptotic hPEBP4 silencing promotes TRAIL-induced apoptosis of human ovarian cancer cells by activating ERK and JNK pathways. Int J Mol Med 18: 505–510. doi: 10.3892/ijmm.18.3.505
[8]
Zhang Y, Wang X, Xiang Z, Li H, Qiu J, et al. (2007) Promotion of cellular migration and apoptosis resistance by a mouse eye-specific phosphatidylethanolamine-binding protein. Int J Mol Med 19: 55–63. doi: 10.3892/ijmm.19.1.55
[9]
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100: 57–70. doi: 10.1016/s0092-8674(00)81683-9
[10]
Meyn RE, Milas L, Ang KK (2009) The role of apoptosis in radiation oncology. Int J Radiat Biol 85: 107–115. doi: 10.1080/09553000802662595
[11]
Fulda S, Debatin KM (2006) Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 25: 4798–4811. doi: 10.1038/sj.onc.1209608
[12]
Qiu J, Yang G, Shen Z, Xie Y, Wang L (2013) hPEBP4 as a predictive marker for the pathological response of rectal cancer to preoperative radiotherapy. Int J Colorectal Dis 28: 241–6. doi: 10.1007/s00384-012-1534-3
[13]
Diehn M, Cho RW, Lobo NA, Kalisky T, Dorie MJ, et al. (2009) Association of reactive oxygen species levels and radioresistance in cancer stem cells. Nature 458: 780–783. doi: 10.1038/nature07733
[14]
Lacher MD, Tiirikainen MI, Saunier EF, Christian C, Anders M, et al. (2006) Transforming growth factor-beta receptor inhibition enhances adenoviral infectability of carcinoma cells via up-regulation of Coxsackie and Adenovirus receptor in conjunction with reversal of epithelial-mesenchymal transition. Cancer Res 66: 1648–1657. doi: 10.1158/0008-5472.can-05-2328
[15]
Cataldi A, Zauli G, Di Pietro R, Castorina S, Rana R (2001) Involvement of the pathway phosphatidylinositol-3-kinase/AKT-1 in the establishment of the survival response to ionizing radiation. Cell Signal 13: 369–375. doi: 10.1016/s0898-6568(01)00147-4
[16]
[16] Rosenzweig KE, Youmell MB, Palayoor ST, Price BD (1997) Radiosensitization of human tumor cells by the phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 correlates with inhibition of DNA-dependent protein kinase and prolonged G2-M delay. Clin Cancer Res 3: 1149–56.
[17]
Tanno S, Yanagawa N, Habiro A, Koizumi K, Nakano Y, et al. (2004) Serine/Threonine Kinase AKT Is Frequently Activated in Human Bile Duct Cancer and Is Associated with Increased Radioresistance. Cancer Research 64: 3486–3490. doi: 10.1158/0008-5472.can-03-1788
[18]
Afanasev I (2011) Reactive Oxygen Species Signaling in Cancer: Comparison with Aging. Aging Diseases 1: 219–230.
[19]
Birgisson H, P？hlman L, Gunnarsson U, Glimelius B (2005) Swedish Rectal Cancer Trial Group (2005) Adverse effects of preoperative radiation therapy for rectal cancer:long-term follow-up of the Swedish Rectal Cancer Trial. J Clin Oncol 23: 8697–8705. doi: 10.1200/jco.2005.02.9017
[20]
Stelzmueller I, Zitt M, Aigner F, Kafka-Ritsch R, J？ger R, et al. (2009) Postoperative morbidity following chemoradiation for locally advanced low rectal cancer. J Gastrointest Surg 13: 657–667. doi: 10.1007/s11605-008-0760-z
[21]
Meyn RE, Milas L, Ang KK (2009) The role of apoptosis in radiation oncology. Int J Radiat Biol 85: 107–115. doi: 10.1080/09553000802662595
[22]
R？del C, Haas J, Groth A, Grabenbauer GG, Sauer R, et al. (2003) Spontaneous and radiationinduced apoptosis in colorectal carcinoma cells with different intrinsic radiosensitivities: Survivin as a radioresistance factor. Int J Radiat Oncol Biol Phys 55: 1341–1347. doi: 10.1016/s0360-3016(02)04618-7
[23]
Pollycove M (2006) Radiobiological basis of low-dose irradiation in prevention and therapy of cancer. Dose Response 5: 26–38. doi: 10.2203/dose-response.06-112.pollycove
[24]
Powell S, McMillan TJ (1990) DNA damage and repair following treatment with ionizing radiation. Radiother Oncol 19: 95–108. doi: 10.1016/0167-8140(90)90123-e
[25]
Ward JF (1985) Biochemistry of DNA lesions. Radiat Res 8: S103–S111. doi: 10.2307/3583517
[26]
Cerutti P (1985) Prooxidant states and tumor promotion. Science 227: 375–381. doi: 10.1126/science.2981433
[27]
Kennedy SG, Kandel ES, Cross TK, Hay N (1999) Akt/Protein kinase B inhibits cell death by preventing the release of cytochrome c from mitochondria. Mol Cell Biol19: 5800–5810.
[28]
Kandel ES, Skeen J, Majewski N, Di Cristofano A, Pandolfi PP, et al. (2002) Activation of Akt/protein kinase B overcomes a G(2)/M cell cycle checkpoint induced by DNA damage. Mol Cell Biol 22: 7831–7841. doi: 10.1128/mcb.22.22.7831-7841.2002
[29]
Albanese C, Wu K, DAmico M, Jarrett C, Joyce D, et al. (2003) IKK regulates mitogenic signaling through transcriptional induction of cyclin D1 via Tcf. Mol Biol Cell 14: 585–599. doi: 10.1091/mbc.02-06-0101
[30]
Oka N, Nakahara S, Takenaka Y, Fukumori T, Hogan V, et al. (2005) Galectin-3 inhibits tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by activating Akt in human bladder carcinoma cells. Cancer Res 65: 7546–7553.
[31]
Leach JK, Van Tuyle G, Lin PS, Schmidt-Ullrich R, Mikkelsen RB (2001) Ionizing radiation-induced, mitochondria-dependent generation of reactive oxygen/nitrogen. Cancer Res 61: 3894–3901.
[32]
Esposito F, Chirico G, Montesano Gesualdi N, Posadas I, Ammendola R, et al. (2003) Protein kinase B activation by reactive oxygen species is independent of tyrosine kinase receptor phosphorylation and requires SRC activity. J Biol Chem 278: 20828–20834. doi: 10.1074/jbc.m211841200
[33]
Shono T, Yokoyama N, Uesaka T, Kuroda J, Takeya R, et al. (2008) Enhanced expression of NADPH oxidase Nox4 in human gliomas and its roles in cell proliferation and survival. Int J Cancer 123: 787–792. doi: 10.1002/ijc.23569
[34]
Sun Y, St Clair DK, Xu Y, Crooks PA, St Clair WH (2010) A NADPH oxidase-dependent redox signaling pathway mediates the selective radiosensitization effect of parthenolide in prostate cancer cells. Cancer Res 70: 2880–2890. doi: 10.1158/0008-5472.can-09-4572
[35]
Mabuchi S, Ohmichi M, Kimura A, Nishio Y, Arimoto Ishida E, et al. (2004) Estrogen inhibits paclitaxel-induced apoptosis via the phosphorylation of apoptosis signal-regulating kinase 1 in human ovarian cancer cell lines. Endocrinology 145: 49–58. doi: 10.1210/en.2003-0792
Liu H, Qiu J, Li N, Chen T, Cao X (2010) Human Phosphatidylethanolamine-binding Protein 4 Promotes Transactivation of Estrogen Receptor α (ERα) in Human Cancer Cells by Inhibiting Proteasome-dependent ERα Degradation via Association with Src. J Biol Chem 285: 21934–21942. doi: 10.1074/jbc.m110.109876
