Abstract
Here we show that pemetrexed-treated mesothelioma cells undergo accelerated senescence. This is characterized by the secretion of proinflammatory and mitogenic cytokines, reminiscent of an SASP (senescence-associated secretory phenotype). Conditioned media from senescent MPM (malignant pleural mesothelioma) cells trigger the emergence of EMT (epithelial-to-mesenchymal)-like, clonogenic and chemoresistant cell subpopulations, expressing high levels of ALDH (aldehyde dehydrogenase) activity (ALDHbright cells). We show by fluorescence-activated cell sorting of purified ALDHbright and ALDHlow cells, that both cell-autonomous and cell-non-autonomous mechanisms converge to maintain the SASP-induced, EMT-like cell subpopulations. Chemoresistant ALDHbright cells exist within primary MPM specimens and enrichment for ALDHbright cells correlates with an earlier tumor onset into NOD/SCID mice. We show that RASv12 expression induces SASP-like changes in untransformed human mesothelial cells, and that p53 ablation increases the effect of RASv12 expression. We identify STAT3 activation as a crucial event downstream to SASP signaling. In fact, small hairpin RNA-mediated ablation of STAT3 deeply attenuates the induction of EMT genes and the increase of ALDHbright cells induced by SASP-cytokines. This strongly affects the chemoresistance of MPM cells in vitro and leads to anticancer effects in vivo.
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Abbreviations
- EMT:
-
epithelial-to-mesenchymal–transition
- MPE:
-
mesothelial peritoneal exudate
- SASP:
-
senescence-associated-secretory-phenotype
- TICs:
-
tumor-initiating–cells
References
Achcar Rde O, Cagle PT, Jagirdar J . (2007). Expression of activated and latent signal transducer and activator of transcription 3 in 303 non-small cell lung carcinomas and 44 malignant mesotheliomas: possible role for chemotherapeutic intervention. Arch Pathol Lab Med 131: 1350–1360.
Adhikari AS, Agarwal N, Iwakuma T . (2011). Metastatic potential of tumor-initiating cells in solid tumors. Front Biosci 16: 1927–1938.
Albonici L, Doldo E, Palumbo C, Orlandi A, Bei R, Pompeo E et al. (2009). Placenta growth factor is a survival factor for human malignant mesothelioma cells. Int J Immunopathol Pharmacol 22: 389–401.
Aroeira LS, Aguilera A, Sanchez-Tomero JA, Bajo MA, del Peso G, Jimenez-Heffernan JA et al. (2007). Epithelial to mesenchymal transition and peritoneal membrane failure in peritoneal dialysis patients: pathologic significance and potential therapeutic interventions. J Am Soc Nephrol 18: 2004–2013.
Bais C, Wu X, Yao J, Yang S, Crawford Y, McCutcheon K et al. (2010). PlGF blockade does not inhibit angiogenesis during primary tumor growth. Cell 141: 166–177.
Ben-Porath I, Weinberg RA . (2004). When cells get stressed: an integrative view of cellular senescence. J Clin Invest 113: 8–13.
Campisi J . (2001). Cellular senescence as a tumor-suppressor mechanism. Trends Cell Biol 11: S27–S31.
Casarsa C, Bassani N, Ambrogi F, Zabucchi G, Boracchi P, Biganzoli E et al. (2011). Epithelial-to-mesenchymal transition, cell polarity and stemness-associated features in malignant pleural mesothelioma. Cancer Lett 302: 136–143.
Charafe-Jauffret E, Ginestier C, Iovino F, Tarpin C, Diebel M, Esterni B et al. (2010). Aldehyde dehydrogenase 1-positive cancer stem cells mediate metastasis and poor clinical outcome in inflammatory breast cancer. Clin Cancer Res 16: 45–55.
Cioce M, Gherardi S, Viglietto G, Strano S, Blandino G, Muti P et al. (2010). Mammosphere-forming cells from breast cancer cell lines as a tool for the identification of CSC-like- and early progenitor-targeting drugs. Cell Cycle 9: 2878–2887.
Coppe JP, Desprez PY, Krtolica A, Campisi J . (2010). The senescence-associated secretory phenotype: the dark side of tumor suppression. Annu Rev Pathol 5: 99–118.
Coppe JP, Patil CK, Rodier F, Sun Y, Munoz DP, Goldstein J et al. (2008). Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol 6: 2853–2868.
Corney DC, Flesken-Nikitin A, Godwin AK, Wang W, Nikitin AY . (2007). MicroRNA-34b and MicroRNA-34c are targets of p53 and cooperate in control of cell proliferation and adhesion-independent growth. Cancer Res 67: 8433–8438.
Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O . (2009). Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc 4: 1798–1806.
Fujino S, Yokoyama A, Kohno N, Hiwada K . (1996). Interleukin 6 is an autocrine growth factor for normal human pleural mesothelial cells. Am J Respir Cell Mol Biol 14: 508–515.
Galffy G, Mohammed KA, Dowling PA, Nasreen N, Ward MJ, Antony VB . (1999). Interleukin 8: an autocrine growth factor for malignant mesothelioma. Cancer Res 59: 367–371.
Hazarika M, White Jr RM, Booth BP, Wang YC, Ham DY, Liang CY et al. (2005). Pemetrexed in malignant pleural mesothelioma. Clin Cancer Res 11: 982–992.
He L, He X, Lim LP, de Stanchina E, Xuan Z, Liang Y et al. (2007). A microRNA component of the p53 tumour suppressor network. Nature 447: 1130–1134.
Hermeking H . (2007). p53 enters the microRNA world. Cancer Cell 12: 414–418.
Hillegass JM, Shukla A, Lathrop SA, MacPherson MB, Beuschel SL, Butnor KJ et al. (2010). Inflammation precedes the development of human malignant mesotheliomas in a SCID mouse xenograft model. Ann N Y Acad Sci 1203: 7–14.
Huang C, Jiang T, Zhu L, Liu J, Cao J, Huang KJ et al. (2011a). STAT3-targeting RNA interference inhibits pancreatic cancer angiogenesis in vitro and in vivo. Int J Oncol 38: 1637–1644.
Huang C, Yang G, Jiang T, Zhu G, Li H, Qiu Z . (2011b). The effects and mechanisms of blockage of STAT3 signaling pathway on IL-6 inducing EMT in human pancreatic cancer cells in vitro. Neoplasma 58: 396–405.
Kim C, Kim DG, Park SH, Hwang YI, Jang SH, Kim CH et al. (2011). Epithelial to mesenchymal transition of mesothelial cells in tuberculous pleurisy. Yonsei Med J 52: 51–58.
Kryczek I, Liu S, Roh M, Vatan L, Szeliga W, Wei S et al. (2011). Expression of aldehyde dehydrogenase and CD133 defines ovarian cancer stem cells. Int J Cancer.
Kubo T, Toyooka S, Tsukuda K, Sakaguchi M, Fukazawa T, Soh J et al. (2011). Epigenetic silencing of microRNA-34b/c lays an important role in the pathogenesis of malignant pleural mesothelioma. Clin Cancer Res 17: 4965–4974.
Laberge RM, Awad P, Campisi J, Desprez PY . (2011). Epithelial-mesenchymal transition induced by senescent fibroblasts. Cancer Microenviron.
Li Q, Yano S, Ogino H, Wang W, Uehara H, Nishioka Y et al. (2007). The therapeutic efficacy of anti vascular endothelial growth factor antibody, bevacizumab, and pemetrexed against orthotopically implanted human pleural mesothelioma cells in severe combined immunodeficient mice. Clin Cancer Res 13: 5918–5925.
Liu S, Ginestier C, Ou SJ, Clouthier SG, Patel SH, Monville F et al. (2011). Breast cancer stem cells are regulated by mesenchymal stem cells through cytokine networks. Cancer Res 71: 614–624.
Ma I, Allan AL . (2011). The role of human aldehyde dehydrogenase in normal and cancer stem cells. Stem Cell Rev 7: 292–306.
Marcato P, Dean CA, Pan D, Araslanova R, Gillis M, Joshi M et al. (2011). Aldehyde dehydrogenase activity of breast cancer stem cells is primarily due to isoform ALDH1A3 and its expression is predictive of metastasis. Stem Cells 29: 32–45.
Metcalf RA, Welsh JA, Bennett WP, Seddon MB, Lehman TA, Pelin K et al. (1992). p53 and Kirsten-ras mutations in human mesothelioma cell lines. Cancer Res 52: 2610–2615.
Mor O, Yaron P, Huszar M, Yellin A, Jakobovitz O, Brok-Simoni F et al. (1997). Absence of p53 mutations in malignant mesotheliomas. Am J Respir Cell Mol Biol 16: 9–13.
Mujoomdar AA, Tilleman TR, Richards WG, Bueno R, Sugarbaker DJ . (2010). Prevalence of in vitro chemotherapeutic drug resistance in primary malignant pleural mesothelioma: Result in a cohort of 203 resection specimens. J Thorac Cardiovasc Surg 140: 352–355.
Oka M, Sakaguchi M, Okada T, Nagai H, Ozaki M, Yoshioka T et al. (2010). Signal transducer and activator of transcription 3 upregulates interleukin-8 expression at the level of transcription in human melanoma cells. Exp Dermatol 19: e50–e55.
Park JS, Kim YS, Jee YK, Myong NH, Lee KY . (2003). Interleukin-8 production in tuberculous pleurisy: role of mesothelial cells stimulated by cytokine network involving tumour necrosis factor-alpha and interleukin-1 beta. Scand J Immunol 57: 463–469.
Pompeo E, Albonici L, Doldo E, Orlandi A, Manzari V, Modesti A et al. (2009). Placenta growth factor expression has prognostic value in malignant pleural mesothelioma. Ann Thorac Surg 88: 426–431.
Rodier F, Kim SH, Nijjar T, Yaswen P, Campisi J . (2005). Cancer and aging: the importance of telomeres in genome maintenance. Int J Biochem Cell Biol 37: 977–990.
Ruco LP, de Laat PA, Matteucci C, Bernasconi S, Sciacca FM, van der Kwast TH et al. (1996). Expression of ICAM-1 and VCAM-1 in human malignant mesothelioma. J Pathol 179: 266–271.
Scarpa S, Giuffrida A, Palumbo C, Coletti A, Cerrito MG, Vasaturo F et al. (2002). Retinoic acid inhibits fibronectin and laminin synthesis and cell migration of human pleural mesothelioma in vitro. Oncol Rep 9: 205–209.
Schindler C, Levy DE, Decker T . (2007). JAK-STAT signaling: from interferons to cytokines. J Biol Chem 282: 20059–20063.
Schramm A, Opitz I, Thies S, Seifert B, Moch H, Weder W et al. (2010). Prognostic significance of epithelial-mesenchymal transition in malignant pleural mesothelioma. Eur J Cardiothorac Surg 37: 566–572.
Sidi R, Pasello G, Opitz I, Soltermann A, Tutic M, Rehrauer H et al. (2011). Induction of senescence markers after neo-adjuvant chemotherapy of malignant pleural mesothelioma and association with clinical outcome: an exploratory analysis. Eur J Cancer 47: 326–332.
Sivertsen S, Hadar R, Elloul S, Vintman L, Bedrossian C, Reich R et al. (2006). Expression of Snail, Slug and Sip1 in malignant mesothelioma effusions is associated with matrix metalloproteinase, but not with cadherin expression. Lung Cancer 54: 309–317.
Suzuki Y, Sakai K, Ueki J, Xu Q, Nakamura T, Shimada H et al. (2010). Inhibition of Met/HGF receptor and angiogenesis by NK4 leads to suppression of tumor growth and migration in malignant pleural mesothelioma. Int J Cancer 127: 1948–1957.
Takeda K, Akira S . (2000). STAT family of transcription factors in cytokine-mediated biological responses. Cytokine Growth Factor Rev 11: 199–207.
Tolnay E, Kuhnen C, Wiethege T, Konig JE, Voss B, Muller KM . (1998). Hepatocyte growth factor/scatter factor and its receptor c-Met are overexpressed and associated with an increased microvessel density in malignant pleural mesothelioma. J Cancer Res Clin Oncol 124: 291–296.
Wang L, Park P, Zhang H, La Marca F, Lin CY . (2011). Prospective identification of tumorigenic osteosarcoma cancer stem cells in OS99-1 cells based on high aldehyde dehydrogenase activity. Int J Cancer 128: 294–303.
Yao J, Wu X, Zhuang G, Kasman IM, Vogt T, Phan V et al. (2011). Expression of a functional VEGFR-1 in tumor cells is a major determinant of anti-PlGF antibodies efficacy. Proc Natl Acad Sci USA 108: 11590–11595.
Young AR, Narita M . (2009). SASP reflects senescence. EMBO Rep 10: 228–230.
Acknowledgements
We thank Ms Tania Merlino (Regina Elena Cancer Institute, Rome) for revising and proofreading the manuscript and Dr Frank Sinicrope (Rochester, MN) for the ShSTAT3 vectors available at ADDGENE (UK). We acknowledge INAIL (Italian Workers’ Compensation Authority) for grant support to GB.
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Canino, C., Mori, F., Cambria, A. et al. SASP mediates chemoresistance and tumor-initiating-activity of mesothelioma cells. Oncogene 31, 3148–3163 (2012). https://doi.org/10.1038/onc.2011.485
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DOI: https://doi.org/10.1038/onc.2011.485