Starvation-induced activation of ATM/Chk2/p53 signaling sensitizes cancer cells to cisplatin

BMC Cancer

Yandong Shi¹  Emanuela Felley-Bosco¹  Thomas M Marti¹  Katrin Orlowski²  Martin Pruschy²  Rolf A Stahel¹ 

[1] Laboratory of Molecular Oncology, Zürich, Switzerland

[2] Department of Radiation Oncology, University Hospital Zürich, University of Zürich, Zürich, Switzerland

关键词: AMPK;    ATM/Chk2/p53 signaling;    cisplatin therapy;    short-term food starvation (STS);    Serum starvation;   

Others  :  1080043
DOI  :  10.1186/1471-2407-12-571

 received in 2012-05-17, accepted in 2012-11-27,  发布年份 2012

【 摘 要 】

Background

Optimizing the safety and efficacy of standard chemotherapeutic agents such as cisplatin (CDDP) is of clinical relevance. Serum starvation in vitro and short-term food starvation in vivo both stress cells by the sudden depletion of paracrine growth stimulation.

Methods

The effects of serum starvation on CDDP toxicity were investigated in normal and cancer cells by assessing proliferation, cell cycle distribution and activation of DNA-damage response and of AMPK, and were compared to effects observed in cells grown in serum-containing medium. The effects of short-term food starvation on CDDP chemotherapy were assessed in xenografts-bearing mice and were compared to effects on tumor growth and/or regression determined in mice with no diet alteration.

Results

We observed that serum starvation in vitro sensitizes cancer cells to CDDP while protecting normal cells. In detail, in normal cells, serum starvation resulted in a complete arrest of cellular proliferation, i.e. depletion of BrdU-incorporation during S-phase and accumulation of the cells in the G0/G1-phase of the cell cycle. Further analysis revealed that proliferation arrest in normal cells is due to p53/p21 activation, which is AMPK-dependent and ATM-independent. In cancer cells, serum starvation also decreased the fraction of S-phase cells but to a minor extent. In contrast to normal cells, serum starvation-induced p53 activation in cancer cells is both AMPK- and ATM-dependent. Combination of CDDP with serum starvation in vitro increased the activation of ATM/Chk2/p53 signaling pathway compared to either treatment alone resulting in an enhanced sensitization of cancer cells to CDDP. Finally, short-term food starvation dramatically increased the sensitivity of human tumor xenografts to cisplatin as indicated not only by a significant growth delay, but also by the induction of complete remission in 60% of the animals bearing mesothelioma xenografts, and in 40% of the animals with lung carcinoma xenografts.

Conclusion

In normal cells, serum starvation in vitro induces a cell cycle arrest and protects from CDDP induced toxicity. In contrast, proliferation of cancer cells is only moderately reduced by serum starvation whereas CDDP toxicity is enhanced. The combination of CDDP treatment with short term food starvation improved outcome in vivo. Therefore, starvation has the potential to enhance the therapeutic index of cisplatin-based therapy.

【 授权许可】

   
2012 Shi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Sanborn RE: Cisplatin Versus Carboplatin in NSCLC: Is There One "Best" Answer? Curr Treat Options Oncol 2009.
• [2]Halazonetis TD, Gorgoulis VG, Bartek J: An oncogene-induced DNA damage model for cancer development. Science 2008, 319:1352-1355.
• [3]Szatrowski TP, Nathan CF: Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 1991, 51:794-798.
• [4]Warburg O: On the origin of cancer cells. Science 1956, 123:309-314.
• [5]Luo J, Solimini NL, Elledge SJ: Principles of cancer therapy: oncogene and non-oncogene addiction. Cell 2009, 136:823-837.
• [6]Raj L, Ide T, Gurkar AU, Foley M, Schenone M, Li X, Tolliday NJ, Golub TR, Carr SA, Shamji AF, et al.: Selective killing of cancer cells by a small molecule targeting the stress response to ROS. Nature 2011, 475:231-234.
• [7]Murga M, Campaner S, Lopez-Contreras AJ, Toledo LI, Soria R, Montana MF, D'Artista L, Schleker T, Guerra C, Garcia E, et al.: Exploiting oncogene-induced replicative stress for the selective killing of Myc-driven tumors. Nat Struct Mol Biol 2011, 18:1331-1335.
• [8]Gjedsted J, Gormsen LC, Nielsen S, Schmitz O, Djurhuus CB, Keiding S, Orskov H, Tonnesen E, Moller N: Effects of a 3-day fast on regional lipid and glucose metabolism in human skeletal muscle and adipose tissue. Acta Physiol (Oxf) 2007, 191:205-216.
• [9]Flemstrom G, Bengtsson MW, Makela K, Herzig KH: Effects of short-term food deprivation on orexin-A-induced intestinal bicarbonate secretion in comparison with related secretagogues. Acta Physiol (Oxf) 2010, 198:373-380.
• [10]Thissen JP, Ketelslegers JM, Underwood LE: Nutritional regulation of the insulin-like growth factors. Endocr Rev 1994, 15:80-101.
• [11]Ahima RS, Prabakaran D, Mantzoros C, Qu D, Lowell B, Maratos-Flier E, Flier JS: Role of leptin in the neuroendocrine response to fasting. Nature 1996, 382:250-252.
• [12]Raff MC: Social controls on cell survival and cell death. Nature 1992, 356:397-400.
• [13]Hanahan D, Weinberg RA: Hallmarks of cancer: the next generation. Cell 2011, 144:646-674.
• [14]Lavin MF, Kozlov S: ATM activation and DNA damage response. Cell Cycle 2007, 6:931-942.
• [15]Guo Z, Kozlov S, Lavin MF, Person MD, Paull TT: ATM activation by oxidative stress. Science 2010, 330:517-521.
• [16]Hickson I, Zhao Y, Richardson CJ, Green SJ, Martin NM, Orr AI, Reaper PM, Jackson SP, Curtin NJ, Smith GC: Identification and characterization of a novel and specific inhibitor of the ataxia-telangiectasia mutated kinase ATM. Cancer Res 2004, 64:9152-9159.
• [17]Bunz F, Dutriaux A, Lengauer C, Waldman T, Zhou S, Brown JP, Sedivy JM, Kinzler KW, Vogelstein B: Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 1998, 282:1497-1501.
• [18]Mihaylova MM, Shaw RJ: The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol 2011, 13:1016-1023.
• [19]Jones RG, Plas DR, Kubek S, Buzzai M, Mu J, Xu Y, Birnbaum MJ, Thompson CB: AMP-activated protein kinase induces a p53-dependent metabolic checkpoint. Mol Cell 2005, 18:283-293.
• [20]Carling D, Zammit VA, Hardie DG: A common bicyclic protein kinase cascade inactivates the regulatory enzymes of fatty acid and cholesterol biosynthesis. FEBS Lett 1987, 223:217-222.
• [21]Handa N, Takagi T, Saijo S, Kishishita S, Takaya D, Toyama M, Terada T, Shirouzu M, Suzuki A, Lee S, et al.: Structural basis for compound C inhibition of the human AMP-activated protein kinase alpha2 subunit kinase domain. Acta Crystallogr D Biol Crystallogr 2011, 67:480-487.
• [22]Lee C, Safdie FM, Raffaghello L, Wei M, Madia F, Parrella E, Hwang D, Cohen P, Bianchi G, Longo VD: Reduced levels of IGF-I mediate differential protection of normal and cancer cells in response to fasting and improve chemotherapeutic index. Cancer Res 2010, 70:1564-1572.
• [23]Lee C, Raffaghello L, Brandhorst S, Safdie FM, Bianchi G, Martin-Montalvo A, Pistoia V, Wei M, Hwang S, Merlino A, et al.: Fasting Cycles Retard Growth of Tumors and Sensitize a Range of Cancer Cell Types to Chemotherapy. Sci Transl Med 2012.
• [24]Raffaghello L, Lee C, Safdie FM, Wei M, Madia F, Bianchi G, Longo VD: Starvation-dependent differential stress resistance protects normal but not cancer cells against high-dose chemotherapy. Proc Natl Acad Sci U S A 2008, 105:8215-8220.
• [25]Sigoillot FD, Berkowski JA, Sigoillot SM, Kotsis DH, Guy HI: Cell cycle-dependent regulation of pyrimidine biosynthesis. J Biol Chem 2003, 278:3403-3409.
• [26]Bester AC, Roniger M, Oren YS, Im MM, Sarni D, Chaoat M, Bensimon A, Zamir G, Shewach DS, Kerem B: Nucleotide deficiency promotes genomic instability in early stages of cancer development. Cell 2011, 145:435-446.
• [27]Yang TY, Chang GC, Chen KC, Hung HW, Hsu KH, Wu CH, Sheu GT, Hsu SL: Pemetrexed induces both intrinsic and extrinsic apoptosis through ataxia telangiectasia mutated/p53-dependent and -independent signaling pathways. Mol Carcinog 2011.
• [28]Vogelzang NJ, Rusthoven JJ, Symanowski J, Denham C, Kaukel E, Ruffie P, Gatzemeier U, Boyer M, Emri S, Manegold C, et al.: Phase III study of pemetrexed in combination with cisplatin versus cisplatin alone in patients with malignant pleural mesothelioma. J Clin Oncol 2003, 21:2636-2644.
• [29]Gridelli C, Maione P, Rossi A, Bareschino MA, Schettino C, Sacco PC, Zeppa R: Pemetrexed in advanced non-small cell lung cancer. Expert Opin Drug Saf 2011, 10:311-317.
• [30]Applegate LA, Luscher P, Tyrrell RM: Induction of heme oxygenase: a general response to oxidant stress in cultured mammalian cells. Cancer Res 1991, 51:974-978.
• [31]Takemura H, Rao VA, Sordet O, Furuta T, Miao ZH, Meng L, Zhang H, Pommier Y: Defective Mre11-dependent activation of Chk2 by ataxia telangiectasia mutated in colorectal carcinoma cells in response to replication-dependent DNA double strand breaks. J Biol Chem 2006, 281:30814-30823.
• [32]Garner KM, Eastman A: Variations in Mre11/Rad50/Nbs1 status and DNA damage-induced S-phase arrest in the cell lines of the NCI60 panel. BMC Cancer 2011, 11(206):201-213.
• [33]Olive PL, Banath JP: Kinetics of H2AX phosphorylation after exposure to cisplatin. Cytometry B Clin Cytom 2009, 76:79-90.
• [34]Lovric MM, Hawkins CJ: TRAIL treatment provokes mutations in surviving cells. Oncogene 2010, 29:5048-5060.
• [35]Towler MC, Hardie DG: AMP-activated protein kinase in metabolic control and insulin signaling. Circ Res 2007, 100:328-341.
• [36]Michalsen A: Prolonged fasting as a method of mood enhancement in chronic pain syndromes: a review of clinical evidence and mechanisms. Curr Pain Headache Rep 2010, 14:80-87.
• [37]Safdie FM, Dorff T, Quinn D, Fontana L, Wei M, Lee C, Cohen P, Longo VD: Fasting and cancer treatment in humans: A case series report. Aging (Albany NY) 2009, 1:988-1007.
• [38]Schmitter D, Lauber B, Fagg B, Stahel RA: Hematopoietic growth factors secreted by seven human pleural mesothelioma cell lines: interleukin-6 production as a common feature. Int J Cancer 1992, 51:296-301.
• [39]Lieber M, Smith B, Szakal A, Nelson-Rees W, Todaro G: A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int J Cancer 1976, 17:62-70.
• [40]Knobel PA, Kotov IN, Felley-Bosco E, Stahel RA, Marti TM: Inhibition of REV3 expression induces persistent DNA damage and growth arrest in cancer cells. Neoplasia 2011, 13:961-970.
• [41]Rheinwald JG, Hahn WC, Ramsey MR, Wu JY, Guo Z, Tsao H, De Luca M, Catricala C, O'Toole KM: A two-stage, p16(INK4A)- and p53-dependent keratinocyte senescence mechanism that limits replicative potential independent of telomere status. Mol Cell Biol 2002, 22:5157-5172.
• [42]Stahel RA, Mabry M, Sabbath K, Speak JA, Bernal SD: Selective cytotoxicity of murine monoclonal antibody LAM2 against human small-cell carcinoma in the presence of human complement: possible use for in vitro elimination of tumor cells from bone marrow. Int J Cancer 1985, 35:587-592.
• [43]Cole SP: Rapid chemosensitivity testing of human lung tumor cells using the MTT assay. Cancer Chemother Pharmacol 1986, 17:259-263.
• [44]Belyanskaya LL, Marti TM, Hopkins-Donaldson S, Kurtz S, Felley-Bosco E, Stahel RA: Human agonistic TRAIL receptor antibodies Mapatumumab and Lexatumumab induce apoptosis in malignant mesothelioma and act synergistically with cisplatin. Mol Cancer 2007, 6:66. BioMed Central Full Text
• [45]Marti TM, Hefner E, Feeney L, Natale V, Cleaver JE: H2AX phosphorylation within the G1 phase after UV irradiation depends on nucleotide excision repair and not DNA double-strand breaks. Proc Natl Acad Sci U S A 2006, 103:9891-9896.
• [46]Tomayko MM, Reynolds CP: Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol 1989, 24:148-154.