Evaluation of premature senescence and senescence biomarkers in carcinoma cells and xenograft mice exposed to single or fractionated irradiation

Kim,Bong  Cho; Yoo,Hee Jung; Lee,Hyung Chul; Kang,Kyoung-Ah; Jung,Seung Hee; Lee,Hae-June; Lee,Minyoung; Park,Seungwoo; Ji,Young-Hoon; Lee,Yun-Sil; Ko,Young-Gyu; Lee,Jae-Seon

doi:10.3892/or.2014.3069

Evaluation of premature senescence and senescence biomarkers in carcinoma cells and xenograft mice exposed to single or fractionated irradiation

Authors:
- Bong Cho Kim
- Hee Jung Yoo
- Hyung Chul Lee
- Kyoung-Ah Kang
- Seung Hee Jung
- Hae-June Lee
- Minyoung Lee
- Seungwoo Park
- Young-Hoon Ji
- Yun-Sil Lee
- Young-Gyu Ko
- Jae-Seon Lee
View Affiliations

Affiliations: Research Center for Radio-senescence, Korea Institute of Radiological and Medical Sciences, Seoul, Republic of Korea, Division of Radiation Effects, Korea Institute of Radiological and Medical Sciences, Seoul, Republic of Korea, Research Center for Radiotherapy, Korea Institute of Radiological and Medical Sciences, Seoul, Republic of Korea, School of Pharmaceutical Sciences, Ewha Womans University, Seoul, Republic of Korea, Division of Life Sciences, Korea University, Seoul, Republic of Korea, Division of Biomedical Sciences, College of Medicine, Inha University, Incheon, Republic of Korea
Published online on: March 7, 2014 https://doi.org/10.3892/or.2014.3069
Pages: 2229-2235

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The purpose of the present study was to elucidate whether premature senescence contributes to the outcome of radiotherapy (RT) and to validate senescence biomarkers in vitro and in vivo. Cultured human cancer cell lines and xenografted mice were exposed to single (SR; 2, 6 or 12 Gy) or fractionated radiation (FR; 3 x 2 Gy or 6 x 2 Gy), and premature senescence was assessed using senescence-associated β-galactosidase (SA-β-Gal) activity, hypophosphorylation of pRb and p21 accumulation. A variety of senescence-associated biomarkers including cathepsin D (CD), the eukaryotic translation elongation factors eEF1A1, eEF1B2, decoy receptor 2 and Dec1 were further validated in vivo or in vitro. We demonstrated the beneficial tumor suppressive role of ionizing radiation (IR)-induced premature senescence in vitro and in vivo. FR inhibited tumor growth via induction of premature senescence as effectively as an equivalent SR dose (≥6 Gy). In addition, CD and eEF1 were valuable biomarkers of cellular senescence in either SR- or RF-exposed carcinoma cells or xenograft mice. Our results suggest that 2 Gy of a conventional RT regime could achieve a better clinical outcome if premature senescence could be increased through an improved understanding of its molecular action mechanism.

View Figures

View References

1	Nomura T, Kinuta M, Hongyo T, Nakajima H and Hatanaka T: Programmed cell death in whole body and organ systems by low dose radiation. J Radiat Res. 33:109–123. 1992. View Article : Google Scholar : PubMed/NCBI
2	Hendry JH, Potten CS and Merritt A: Apoptosis induced by high- and low-LET radiations. Radiat Environ Biophys. 34:59–62. 1995. View Article : Google Scholar : PubMed/NCBI
3	Wang Y, Liu L, Pazhanisamy SK, Li H, Meng A and Zhou D: Total body irradiation causes residual bone marrow injury by induction of persistent oxidative stress in murine hematopoietic stem cells. Free Radic Biol Med. 48:348–356. 2010. View Article : Google Scholar : PubMed/NCBI
4	Bruskov VI, Karp OE, Garmash SA, Shtarkman IN, Chernikov AV and Gudkov SV: Prolongation of oxidative stress by long-lived reactive protein species induced by X-ray radiation and their genotoxic action. Free Radic Res. 46:1280–1290. 2012. View Article : Google Scholar : PubMed/NCBI
5	Gewirtz DA, Holt SE and Elmore LW: Accelerated senescence: an emerging role in tumor cell response to chemotherapy and radiation. Biochem Pharmacol. 76:947–957. 2008. View Article : Google Scholar : PubMed/NCBI
6	Collado M and Serrano M: Senescence in tumours: evidence from mice and humans. Nat Rev Cancer. 10:51–57. 2010. View Article : Google Scholar : PubMed/NCBI
7	Nardella C, Clohessy JG, Alimonti A and Pandolfi PP: Pro-senescence therapy for cancer treatment. Nat Rev Cancer. 11:503–511. 2011. View Article : Google Scholar : PubMed/NCBI
8	Ben-Porath I and Weinberg RA: The signals and pathways activating cellular senescence. Int J Biochem Cell Biol. 37:961–976. 2005. View Article : Google Scholar : PubMed/NCBI
9	Ohtani N, Mann DJ and Hara E: Cellular senescence: its role in tumor suppression and aging. Cancer Sci. 100:792–797. 2009. View Article : Google Scholar : PubMed/NCBI
10	Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, et al: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA. 92:9363–9367. 1995. View Article : Google Scholar : PubMed/NCBI
11	Debacq-Chainiaux F, Erusalimsky JD, Campisi J and Toussaint O: Protocols to detect senescence-associated beta-galactosidase (SA-βgal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc. 4:1798–1806. 2009.PubMed/NCBI
12	Chen Z, Trotman LC, Shaffer D, Lin HK, Dotan ZA, Niki M, Koutcher JA, Scher HI, Ludwig T, Gerald W, Cordon-Cardo C and Pandolfi PP: Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. Nature. 436:725–730. 2005. View Article : Google Scholar : PubMed/NCBI
13	Collado M, Gil J, Efeyan A, Guerra C, Schuhmacher AJ, Barradas M, Benguría A, Zaballos A, Flores JM, Barbacid M, Beach D and Serrano M: Tumour biology: senescence in premalignant tumours. Nature. 436:6422005. View Article : Google Scholar : PubMed/NCBI
14	Dankort D, Filenova E, Collado M, Serrano M, Jones K and McMahon M: A new mouse model to explore the initiation, progression, and therapy of BRAFV600E-induced lung tumors. Genes Dev. 21:379–384. 2007. View Article : Google Scholar : PubMed/NCBI
15	Byun HO, Han NK, Lee HJ, Kim KB, Ko YG, Yoon G, Lee YS, Hong SI and Lee JS: Cathepsin D and eukaryotic translation elongation factor 1 as promising markers of cellular senescence. Cancer Res. 69:4638–4647. 2009. View Article : Google Scholar : PubMed/NCBI
16	Spallarossa P, Altieri P, Aloi C, Garibaldi S, Barisione C, Ghigliotti G, Fugazza G, Barsotti A and Brunelli C: Doxorubicin induces senescence or apoptosis in rat neonatal cardiomyocytes by regulating the expression levels of the telomere binding factors 1 and 2. Am J Physiol Heart Circ Physiol. 297:H2169–H2181. 2009. View Article : Google Scholar : PubMed/NCBI
17	Nübel T, Damrot J, Roos WP, Kaina B and Fritz G: Lovastatin protects human endothelial cells from killing by ionizing radiation without impairing induction and repair of DNA double-strand breaks. Clin Cancer Res. 12:933–939. 2006.PubMed/NCBI
18	Rübe CE, Fricke A, Wendorf J, Stützel A, Kühne M, Ong MF, Lipp P and Rübe C: Accumulation of DNA double-strand breaks in normal tissues after fractionated irradiation. Int J Radiat Oncol Biol Phys. 76:1206–1213. 2010.PubMed/NCBI
19	Kalderon N, Xu S, Koutcher JA and Fuks Z: Fractionated radiation facilitates repair and functional motor recovery after spinal cord transection in rat. Brain Res. 904:199–207. 2001. View Article : Google Scholar : PubMed/NCBI
20	Dearnaley DP, Khoo VS, Norman AR, Meyer L, Nahum A, Tait D, Yarnold J and Horwich A: Comparison of radiation side-effects of conformal and conventional radiotherapy in prostate cancer: a randomised trial. Lancet. 353:267–272. 1999. View Article : Google Scholar : PubMed/NCBI
21	Munshi A and Budrukkar A: Hypofractionated radiation therapy in breast cancer: a revolutionary breakthrough or a long way to go? J Clin Oncol. 25:458–459. 2007. View Article : Google Scholar : PubMed/NCBI
22	Collado M and Serrano M: The power and the promise of oncogene-induced senescence markers. Nat Rev Cancer. 6:472–476. 2006. View Article : Google Scholar : PubMed/NCBI
23	Suzuki M and Boothman DA: Stress-induced premature senescence (SIPS) - influence of SIPS on radiotherapy. J Radiat Res. 49:105–112. 2008. View Article : Google Scholar : PubMed/NCBI
24	Lee JJ, Lee JH, Ko YG, Hong SI and Lee JS: Prevention of premature senescence requires JNK regulation of Bcl-2 and reactive oxygen species. Oncogene. 29:561–575. 2010. View Article : Google Scholar : PubMed/NCBI
25	Lee JJ, Kim BC, Park MJ, Lee YS, Kim YN, Lee BL and Lee JS: PTEN status switches cell fate between premature senescence and apoptosis in glioma exposed to ionizing radiation. Cell Death Differ. 18:666–677. 2011. View Article : Google Scholar : PubMed/NCBI
26	Chen WS, Yu YC, Lee YJ, Chen JH, Hsu HY and Chiu SJ: Depletion of securin induces senescence after irradiation and enhances radiosensitivity in human cancer cells regardless of functional p53 expression. Int J Radiat Oncol Biol Phys. 77:566–574. 2010. View Article : Google Scholar : PubMed/NCBI
27	Jackson JG, Post SM and Lozano G: Regulation of tissue- and stimulus-specific cell fate decisions by p53 in vivo. J Pathol. 223:127–136. 2011. View Article : Google Scholar : PubMed/NCBI
28	Haugstetter AM, Loddenkemper C, Lenze D, Gröne J, Standfuss C, Petersen I, Dörken B and Schmitt CA: Cellular senescence predicts treatment outcome in metastasised colorectal cancer. Br J Cancer. 103:505–509. 2010. View Article : Google Scholar : PubMed/NCBI
29	Chang BD, Broude EV, Dokmanovic M, Zhu H, Ruth A, Xuan Y, Kandel ES, Lausch E, Christov K and Roninson IB: A senescence-like phenotype distinguishes tumor cells that undergo terminal proliferation arrest after exposure to anticancer agents. Cancer Res. 59:3761–3767. 1999.
30	Ruth AC and Roninson IB: Effects of the multidrug transporter P-glycoprotein on cellular responses to ionizing radiation. Cancer Res. 60:2576–2578. 2000.PubMed/NCBI
31	Podtcheko A, Ohtsuru A, Namba H, Saenko V, Starenki D, Palona I, Sedliarou I, Rogounovitch T and Yamashita S: Inhibition of ABL tyrosine kinase potentiates radiation-induced terminal growth arrest in anaplastic thyroid cancer cells. Radiat Res. 165:35–42. 2006. View Article : Google Scholar
32	Efimova EV, Mauceri HJ, Golden DW, Labay E, Bindokas VP, Darga TE, Chakraborty C, Barreto-Andrade JC, Crawley C, Sutton HG, Kron SJ and Weichselbaum RR: Poly(ADP-ribose) polymerase inhibitor induces accelerated senescence in irradiated breast cancer cells and tumors. Cancer Res. 70:6277–6282. 2010. View Article : Google Scholar : PubMed/NCBI
33	DeMasters GA, Gupta MS, Jones KR, Cabot M, Wang H, Gennings C, Park M, Bratland A, Ree AH and Gewirtz DA: Potentiation of cell killing by fractionated radiation and suppression of proliferative recovery in MCF-7 breast tumor cells by the Vitamin D3 analog EB 1089. J Steroid Biochem Mol Biol. 92:365–374. 2004. View Article : Google Scholar : PubMed/NCBI
34	Pearson M, Carbone R, Sebastiani C, Cioce M, Fagioli M, Saito S, Higashimoto Y, Appella E, Minucci S, Pandolfi PP and Pelicci PG: PML regulates p53 acetylation and premature senescence induced by oncogenic Ras. Nature. 406:207–210. 2000. View Article : Google Scholar : PubMed/NCBI