Survivin S81A Enhanced TRAIL's Activity in Inducing Apoptosis

Ferry Sandra, Roya Khosravi-Far


BACKGROUND: Survivin is rarely expressed in normal healthy adult tissues, however, it is upregulated in the majority of cancers. Survivin, which belongs to IAPs family, has been widely reported to protect cells from apoptosis by inhibiting caspases pathway. Survivin’s mitotic activity is modulated by many kinases, and its phosphor status can also influence its ability to inhibit apoptosis. There are several important survivin’s phosphorylation sites, such as S20 and T34. We have continued our investigation on other potential survivin’s phosphorylation sites that could be important site for regulating survivin’s cyto-protection.

METHODS: By assuming that S81 could be a potential target to modify activity of survivin, wild-type survivin (Survivin), antisense survivin (Survivin-AS), mutated-survivin Thr34Ala (Survivin-T34A) and mutated-survivin Ser81Ala (Survivin-S81A) were constructed and inserted into pMSCV-IRES-GFP vector with cytomegalovirus (CMV) promoter. Each retroviral product was produced in BOSC23 cells. LY294002 pretreatment and TRAIL treatment along with infection of retroviral products were performed in murine fibrosarcoma L929 cells. For analysis, flow cytometric apoptosis assay and western blot were performed.

RESULTS: In our present study, survivin for providing cytoprotection was regulated by PI3K. The results showed that LY294002, an inhibitor of PI3K, effectively suppressed survivin-modulated cytoprotection in a TRAIL-induced apoptotic model. In addition, mutated survivin S81A showed marked suppression on survivin’s cytoprotection. Along with that, TRAIL’s apoptotic activity was enhanced for inducing apoptosis.

CONCLUSION: We suggested that survivin could inhibit apoptosis through PI3K and S81A could be another potential target in order to inhibit Survivin-modulated cytoprotection as well as to sensitize efficacy of TRAIL or other related apoptotic inducers.

KEYWORDS: apoptosis, survivin, TRAIL, S81A, L929, LY294002

Full Text:



Altieri DC. New wirings in the survivin networks. Oncogene. 2008; 27: 6276-84, CrossRef.

Ryan BM, O’Donovan N, Duffy MJ. Survivin: A new target for anti-cancer therapy. Cancer Treatment Reviews. 2009; 35: 553-62, CrossRef.

Altieri DC. Survivin and IAP proteins in cell-death mechanisms. Biochem J. 2010; 430: 199-205, CrossRef.

Mita AC, Mita MM, Nawrocki ST, Giles FJ. Survivin: key regulator of mitosis and apoptosis and novel target for cancer therapeutics. Clin Cancer Res. 2008; 14: 5000-5, CrossRef.

Barrett RMA, Colnaghi R, Wheatley SP. Threonine 48 in the BIR domain of survivin is critical to its mitotic and anti-apoptotic activities and can be phosphorylated by CK2 in vitro. Cell Cycle. 2011; 10: 538-48, CrossRef.

Siegelin MD, Reuss DE, Habel A, Rami A, von Deimling A. Quercetin promotes degradation of survivin and thereby enhances death-receptor-mediated apoptosis in glioma cells. Neuro Oncol. 2008; 11: 122-31, CrossRef.

Gaiser T, Becker MR, Habel A, Reuss DE, Ehemann V, Rami A, et al. TRAIL-mediated apoptosis in malignant glioma cells is augmented by celecoxib through proteasomal degradation of survivin. Neurosci Lett. 2008; 442: 109-13, CrossRef.

Schultze K, Böck B, Eckert A, Oevermann L, Ramacher D, Wiestler O, et al. Troglitazone sensitizes tumor cells to TRAIL-induced apoptosis via down-regulation of FLIP and Survivin. Apoptosis. 2006; 11: 1503-12, CrossRef.

Sung B, Park B, Yadav VR, Aggarwal BB. Celastrol, a triterpene, enhances TRAIL-induced apoptosis through the down-regulation of cell survival proteins and up-regulation of death receptors. J Biol Chem. 2010; 285: 11498-507, CrossRef.

Zhang HY, Du ZX, Liu BQ, Gao YY, Meng X, Guan Y, et al. Tunicamycin enhances TRAIL-induced apoptosis by inhibition of cyclin D1 and the subsequent downregulation of survivin. Exp Mol Med. 2009; 41: 362-9, CrossRef.

Niu TK, Cheng Y, Ren X, Yang JM. Interaction of Beclin 1 with survivin regulates sensitivity of human glioma cells to TRAIL-induced apoptosis. FEBS Letters. 2010; 584: 3519-24, CrossRef.

Kim JY, Kim EH, Kim SU, Kwon TK, Choi KS. Capsaicin sensitizes malignant glioma cells to TRAIL-mediated apoptosis via DR5 upregulation and survivin downregulation. Carcinogenesis. 2009; 31: 367-75, CrossRef.

Kim EH, Kim HS, Kim SU, Noh EJ, Lee JS, Choi KS. Sodium butyrate sensitizes human glioma cells to TRAIL-mediated apoptosis through inhibition of Cdc2 and the subsequent downregulation of survivin and XIAP. Oncogene. 2005; 24: 6877-89, CrossRef.

Lu M, Strohecker A, Chen F, Kwan T, Bosman J, Jordan VC, et al. Aspirin sensitizes cancer cells to TRAIL-induced apoptosis by reducing survivin levels. Clin Cancer Res. 2008; 14: 3168-76, CrossRef.

Aspe JR, Wall NR. Survivin-T34A: molecular mechanism and therapeutic potential. Onco Targets Ther. 2010; 3: 247-54, CrossRef.

Sandra F, Matsuda M, Yoshida H, Hirata M. Inositol hexakisphosphate blocks tumor cell growth by activating apoptotic machinery as well as by inhibiting the Akt/NFkappaB-mediated cell survival pathway. Carcinogenesis. 2002; 23: 2031-41, CrossRef.

Altieri DC. Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer. 2008; 8: 61-70, CrossRef.

Eckelman BP, Salvesen GS, Scott FL. Human inhibitor of apoptosis proteins: why XIAP is the black sheep of the family. EMBO rep. 2006; 7: 988-94, CrossRef.

Dohi T, Beltrami E, Wall NR, Plescia J, Altieri DC. Mitochondrial survivin inhibits apoptosis and promotes tumorigenesis. J Clin Invest. 2004; 114: 1117-27, CrossRef.

Marusawa H. HBXIP functions as a cofactor of survivin in apoptosis suppression. EMBO J. 2003; 22: 2729-40, CrossRef.

Verhagen AM, Ekert PG, Pakusch M, Silke J, Connolly LM, Reid GE, et al. Identification of DIABLO, a mammalian protein that promotes apoptosis by binding to and antagonizing IAP proteins. Cell. 2000; 102: 43-53, CrossRef.

Vaira V, Lee CW, Goel HL, Bosari S, Languino LR, Altieri DC. Regulation of survivin expression by IGF-1/mTOR signaling. Oncogene. 2006; 26: 2678-84, CrossRef.

O’Connor DS, Grossman D, Plescia J, Li F, Zhang H, Villa A, et al. Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. Proc Natl Acad Sci. 2000; 9: 13103-7, CrossRef.

O’Connor DS, Wall NR, Porter AC, Altieri DC. A p34cdc2 survival checkpoint in cancer. Cancer Cell. 2002; 2: 43-54, CrossRef.

Wheatley SP, Henzing AJ, Dodson H, Khaled W, Earnshaw WC. Aurora-B phosphorylation in vitro identifies a residue of survivin that is essential for its localization and binding to inner centromere protein (INCENP) in vivo. J Biol Chem. 2003; 279: 5655-60, CrossRef.

Wheatley SP, Barrett RM, Andrews PD, Medema RH, Morley SJ, Swedlow JR, et al. Phosphorylation by aurora-B negatively regulates survivin function during mitosis. Cell Cycle. 2007; 6: 1220-30, CrossRef.

Colnaghi R, Wheatley SP. Liaisons between survivin and Plk1 during cell division and cell death. J Biol Chem. 2010; 285: 22592-604, CrossRef.

Barrett RMA, Osborne TP, Wheatley SP. Phosphorylation of survivin at threonine 34 inhibits its mitotic function and enhances its cytoprotective activity. Cell Cycle. 2009; 8: 278-83,CrossRef.

Wall NR, O'Connor DS, Plescia J, Pommier Y, Altieri DC. Suppression of survivin phosphorylation on Thr34 by flavopiridol enhances tumor cell apoptosis. Cancer Res. 2003; 63: 230-5, PMID.


Indexed by:






The Prodia Education and Research Institute