Abstract
These investigations demonstrate that expression of the inhibitor of apoptosis family member, survivin, is dramatically increased during immortalization of nontransformed human fibroblasts that were transduced with telomerase reverse transcriptase (hTERT). Expression of survivin in immortalized fibroblasts peaked during G2/M phase of the cell cycle. However, the upregulation of survivin was dissociated from the rate of proliferation and proportion of G2/M cells. Depletion of survivin from immortal fibroblasts increased sensitivity to stress-induced apoptosis and resulted in an accumulation of cells with 4N DNA content. Conversely, overexpression of survivin in mortal fibroblasts conferred resistance to apoptosis. In contrast, very low levels of survivin in proliferating parental fibroblasts had no bearing on sensitivity to apoptosis. The upregulation of survivin did not appear to be a direct consequence of hTERT transduction. However, repression of hTERT resulted in the rapid downregulation of survivin in telomerase-immortalized fibroblasts and tumor cell lines, but not in cells immortalized via an Alternative Lengthening of Telomeres mechanism. These results have important therapeutic implications, as telomerase and survivin are both broadly expressed in human cancers. Selection during the immortalization process for cells expressing high levels of survivin may account for the abundance of survivin in diverse tumor types.
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References
Adida C, Crotty PL, McGrath J, Berrebi D, Diebold J, Altieri DC . (1998). Developmentally regulated expression of the novel cancer anti-apoptosis gene survivin in human and mouse differentiation. Am J Pathol 152: 43–49.
Ambrosini G, Adida C, Altieri DC . (1997). A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 3: 917–921.
Arora V, Cheung HH, Plenchette S, Micali OC, Liston P, Korneluk RG . (2007). Degradation of survivin by the X-linked inhibitor of apoptosis (XIAP)-XAF1 complex. J Biol Chem 282: 26202–26209.
Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu CP, Morin GB et al. (1998). Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349–352.
Carvalho A, Carmena M, Sambade C, Earnshaw WC, Wheatley SP . (2003). Survivin is required for stable checkpoint activation in taxol-treated HeLa cells. J Cell Sci 116: 2987–2998.
Cohen SB, Graham ME, Lovrecz GO, Bache N, Robinson PJ, Reddel RR . (2007). Protein composition of catalytically active human telomerase from immortal cells. Science 315: 1850–1853.
Colgin LM, Wilkinson C, Englezou A, Kilian A, Robinson MO, Reddel RR . (2000). The hTERTalpha splice variant is a dominant negative inhibitor of telomerase activity. Neoplasia 2: 426–432.
Colnaghi R, Connell CM, Barrett RM, Wheatley SP . (2006). Separating the anti-apoptotic and mitotic roles of survivin. J Biol Chem 281: 33450–33456.
Dohi T, Okada K, Xia F, Wilford CE, Samuel T, Welsh K et al. (2004). An IAP-IAP complex inhibits apoptosis. J Biol Chem 279: 34087–34090.
Fukuda S, Pelus LM . (2001). Regulation of the inhibitor-of-apoptosis family member survivin in normal cord blood and bone marrow CD34(+) cells by hematopoietic growth factors: implication of survivin expression in normal hematopoiesis. Blood 98: 2091–2100.
Gorbunova V, Seluanov A, Pereira-Smith OM . (2002). Expression of human telomerase (hTERT) does not prevent stress-induced senescence in normal human fibroblasts but protects the cells from stress-induced apoptosis and necrosis. J Biol Chem 277: 38540–38549.
Gurbuxani S, Xu Y, Keerthivasan G, Wickrema A, Crispino JD . (2005). Differential requirements for survivin in hematopoietic cell development. Proc Natl Acad Sci USA 102: 11480–11485.
Hiyama K, Hirai Y, Kyoizumi S, Akiyama M, Hiyama E, Piatyszek MA et al. (1995). Activation of telomerase in human lymphocytes and hematopoietic progenitor cells. J Immunol 155: 3711–3715.
Hoffman WH, Biade S, Zilfou JT, Chen J, Murphy M . (2002). Transcriptional repression of the anti-apoptotic survivin gene by wild type p53. J Biol Chem 277: 3247–3257.
Huschtscha LI, Holliday R . (1983). Limited and unlimited growth of SV40-transformed cells from human diploid MRC-5 fibroblasts. J Cell Sci 63: 77–99.
Jiang Y, Saavedra HI, Holloway MP, Leone G, Altura RA . (2004). Aberrant regulation of survivin by the RB/E2F family of proteins. J Biol Chem 279: 40511–40520.
Kiyono T, Foster SA, Koop JI, McDougall JK, Galloway DA, Klingelhutz AJ . (1998). Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature 396: 84–88.
Knauer SK, Bier C, Habtemichael N, Stauber RH . (2006). The Survivin-Crm1 interaction is essential for chromosomal passenger complex localization and function. EMBO Rep 7: 1259–1265.
Knauer SK, Kramer OH, Knosel T, Engels K, Rodel F, Kovacs AF et al. (2007). Nuclear export is essential for the tumor-promoting activity of survivin. FASEB J 21: 207–216.
Lens SM, Wolthuis RM, Klompmaker R, Kauw J, Agami R, Brummelkamp T et al. (2003). Survivin is required for a sustained spindle checkpoint arrest in response to lack of tension. EMBO J 22: 2934–2947.
Li F, Ackermann EJ, Bennett CF, Rothermel AL, Plescia J, Tognin S et al. (1999). Pleiotropic cell-division defects and apoptosis induced by interference with survivin function. Nat Cell Biol 1: 461–466.
Li F, Ambrosini G, Chu EY, Plescia J, Tognin S, Marchisio PC et al. (1998). Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 396: 580–584.
MacKenzie KL, Franco S, May C, Sadelain M, Moore MA . (2000). Mass cultured human fibroblasts overexpressing hTERT encounter a growth crisis following an extended period of proliferation. Exp Cell Res 259: 336–350.
MacKenzie KL, Franco S, Naiyer AJ, May C, Sadelain M, Rafii S et al. (2002). Multiple stages of malignant transformation of human endothelial cells modelled by co-expression of telomerase reverse transcriptase, SV40 T antigen and oncogenic N-ras. Oncogene 21: 4200–4211.
Masutomi K, Possemato R, Wong JM, Currier JL, Tothova Z, Manola JB et al. (2005). The telomerase reverse transcriptase regulates chromatin state and DNA damage responses. Proc Natl Acad Sci USA 102: 8222–8227.
Mesri M, Wall NR, Li J, Kim RW, Altieri DC . (2001). Cancer gene therapy using a survivin mutant adenovirus. J Clin Invest 108: 981–990.
Milyavsky M, Shats I, Cholostoy A, Brosh R, Buganim Y, Weisz L et al. (2007). Inactivation of myocardin and p16 during malignant transformation contributes to a differentiation defect. Cancer Cell 11: 133–146.
Milyavsky M, Shats I, Erez N, Tang X, Senderovich S, Meerson A et al. (2003). Prolonged culture of telomerase-immortalized human fibroblasts leads to a premalignant phenotype. Cancer Res 63: 7147–7157.
Mirza A, McGuirk M, Hockenberry TN, Wu Q, Ashar H, Black S et al. (2002). Human survivin is negatively regulated by wild-type p53 and participates in p53-dependent apoptotic pathway. Oncogene 21: 2613–2622.
Pennati M, Folini M, Zaffaroni N . (2007). Targeting survivin in cancer therapy: fulfilled promises and open questions. Carcinogenesis 28: 1133–1139.
Raj D, Liu T, Samadashwily G, Li F, Grossman D . (2008). Survivin repression by p53, Rb and E2F2 in normal human melanocytes. Carcinogenesis 29: 194–201.
Reddel RR . (2000). The role of senescence and immortalization in carcinogenesis. Carcinogenesis 21: 477–484.
Rodel F, Hoffmann J, Distel L, Herrmann M, Noisternig T, Papadopoulos T et al. (2005). Survivin as a radioresistance factor, and prognostic and therapeutic target for radiotherapy in rectal cancer. Cancer Res 65: 4881–4887.
Schuller CE, Jankowski K, Mackenzie KL . (2007). Telomere length of cord blood-derived CD34(+) progenitors predicts erythroid proliferative potential. Leukemia 21: 983–991.
Stewart SA, Weinberg RA . (2006). Telomeres: cancer to human aging. Annu Rev Cell Dev Biol 22: 531–557.
Taylor LM, James A, Schuller CE, Brce J, Lock RB, Mackenzie KL . (2004). Inactivation of p16INK4a, with retention of pRB and p53/p21cip1 function, in human MRC5 fibroblasts that overcome a telomere-independent crisis during immortalization. J Biol Chem 279: 43634–43645.
Uren AG, Wong L, Pakusch M, Fowler KJ, Burrows FJ, Vaux DL et al. (2000). Survivin and the inner centromere protein INCENP show similar cell-cycle localization and gene knockout phenotype. Curr Biol 10: 1319–1328.
Verhagen AM, Coulson EJ, Vaux DL . (2001). Inhibitor of apoptosis proteins and their relatives: IAPs and other BIRPs. Genome Biol 2: REVIEWS3009.
Wang J, Yang L, Yang J, Kuropatwinski K, Wang W, Liu XQ et al. (2008). Transforming growth factor beta induces apoptosis through repressing the phosphoinositide 3-kinase/AKT/survivin pathway in colon cancer cells. Cancer Res 68: 3152–3160.
Wang Z, Sampath J, Fukuda S, Pelus LM . (2005). Disruption of the inhibitor of apoptosis protein survivin sensitizes Bcr-abl-positive cells to STI571-induced apoptosis. Cancer Res 65: 8224–8232.
Weinstein IB, Joe A . (2008). Oncogene addiction. Cancer Res 68: 3077–3080; discussion 3080.
Wen VW, Wu K, Baksh S, Hinshelwood RA, Lock RB, Clark SJ et al. (2006). Telomere-driven karyotypic complexity concurs with p16INK4a inactivation in TP53-competent immortal endothelial cells. Cancer Res 66: 10691–10700.
Wright WE, Piatyszek MA, Rainey WE, Byrd W, Shay JW . (1996). Telomerase activity in human germline and embryonic tissues and cells. Dev Genet 18: 173–179.
Yang D, Welm A, Bishop JM . (2004). Cell division and cell survival in the absence of survivin. Proc Natl Acad Sci USA 101: 15100–15105.
Yang J, Song K, Krebs TL, Jackson MW, Danielpour D . (2008). Rb/E2F4 and Smad2/3 link survivin to TGF-beta-induced apoptosis and tumor progression. Oncogene 27: 5326–5338.
Zhang M, Yang J, Li F . (2006). Transcriptional and post-transcriptional controls of survivin in cancer cells: novel approaches for cancer treatment. J Exp Clin Cancer Res 25: 391–402.
Acknowledgements
We thank Dr John D Crispino (The Ben May Institute for Cancer Research, IL) for providing us with retroviral MIGR-Survivin, Dr Dario C Altieri (University of Massachusetts Medical School, MA, USA) for providing the Adv-T34A adenoviral vector, Dr Gary Nolan (Stanford University, CA, USA) for Phoenix A packaging cells and Dr Lily Huschtscha, Jane Noble and Dr Paul Bonnefin (Children's Medical Research Institute, New South Wales Australia) for providing immortalized cells. We also thank Laura Veas for proofreading this article. This work was supported by funds from the National Health and Medical Research Council (ID no. 510378 and 568704; KLM), New South Wales Cancer Council (KLM) and Cancer Institute New South Wales (KLM). Children's Cancer Institute Australia is affiliated with the University of New South Wales and Sydney Children's Hospital.
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Yuan, J., Yang, BP., Zhong, ZH. et al. Upregulation of survivin during immortalization of nontransformed human fibroblasts transduced with telomerase reverse transcriptase. Oncogene 28, 2678–2689 (2009). https://doi.org/10.1038/onc.2009.136
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DOI: https://doi.org/10.1038/onc.2009.136
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