Cancer-associated fibroblasts from human NSCLC survive ablative doses of radiation but their invasive capacity is reduced

Radiation Oncology

Iñigo Martinez-Zubiaurre³  Anthony Chalmers⁵  Roy Bremnes³  Lill-Tove Busund⁶  Ruth H Paulssen³  Kristian Bartnes²  Bjørn T Moe⁶  Jan Olof Winberg¹  Vivian Berg³  Ingvild Pettersen³  Turid Hellevik⁴ 

[1] Department of Medical Biology, University of Tromsø, 9037 Tromsø, Norway;Department of Cardiothoracic and Vascular Surgery, University Hospital of Northern Norway, 9038 Tromsø, Norway;Department of Clinical Medicine, University of Tromsø, 9037, Tromsø, Norway;Department of Oncology, University Hospital of Northern Norway, 9038 Tromsø, Norway;Institute of Cancer Sciences, University of Glasgow, Glasgow, UK;Department of Clinical Pathology, University Hospital of Northern Norway, 9038 Tromsø, Norway

关键词: Focal adhesion;    Integrins;    Invasion;    Ablative radiation;    Cancer-associated fibroblasts;   

Others  :  1160857
DOI  :  10.1186/1748-717X-7-59

 received in 2011-12-13, accepted in 2012-04-13,  发布年份 2012

【 摘 要 】

Background

Cancer-Associated Fibroblasts (CAFs) are significant components of solid malignancies and play central roles in cancer sustainability, invasion and metastasis. In this study we have investigated the invasive capacity and matrix remodelling properties of human lung CAFs after exposure to ablative doses of ionizing radiation (AIR), equivalent to single fractions delivered by stereotactic ablative radiotherapy (SART) for medically inoperable stage-I/II non-small-cell lung cancers.

Methods

CAFs were isolated from lung tumour specimens from 16 donors. Initially, intrinsic radiosensitivity was evaluated by checking viability and extent of DNA-damage response (DDR) at different radiation doses. The migrative and invasive capacities of CAFs were thereafter determined after a sub-lethal single radiation dose of 18 Gy. To ascertain the mechanisms behind the altered invasive capacity of cells, expression of matrix metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs) were measured in the conditioned media several days post-irradiation, along with expression of cell surface integrins and dynamics of focal contacts by vinculin-staining.

Results

Exposing CAFs to 1 × 18 Gy resulted in a potent induction of multiple nuclear DDR foci (> 9/cell) with little resolution after 120 h, induced premature cellular senescence and inhibition of the proliferative, migrative and invasive capacity. AIR promoted MMP-3 and inhibited MMP-1 appearance to some extent, but did not affect expression of other major MMPs. Furthermore, surface expression of integrins α2, β1 and α5 was consistently enhanced, and a dramatic augmentation and redistribution of focal contacts was observed.

Conclusions

Our data indicate that ablative doses of radiation exert advantageous inhibitory effects on the proliferative, migratory and invasive capacity of lung CAFs. The reduced motility of irradiated CAFs might be a consequence of stabilized focal contacts via integrins.

【 授权许可】

   
2012 Hellevik et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Heinzerling JH, Kavanagh B, Timmerman RD: Stereotactic ablative radiation therapy for primary lung tumors. Cancer J 2011, 17(1):28-32.
• [2]Palma D, Senan S: Stereotactic radiation therapy: changing treatment paradigms for stage I nonsmall cell lung cancer. Curr Opin Oncol 2011, 23(2):133-139.
• [3]Kavanagh BD, Miften M, Rabinovitch RA: Advances in treatment techniques: stereotactic body radiation therapy and the spread of hypofractionation. Cancer J 2011, 17(3):177-181.
• [4]Bissell MJ, Radisky D: Putting tumours in context. Nat Rev Cancer 2001, 1(1):46-54.
• [5]Bhowmick NA, Neilson EG, Moses HL: Stromal fibroblasts in cancer initiation and progression. Nature 2004, 432(7015):332-337.
• [6]Kalluri R, Zeisberg M: Fibroblasts in cancer. Nat Rev Cancer 2006, 6(5):392-401.
• [7]Gaggioli C, Hooper S, Hidalgo-Carcedo C, Grosse R, Marshall JF, Harrington K, Sahai E: Fibroblast-led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells. Nat Cell Biol 2007, 9(12):1392-1400.
• [8]Chantrain CF, Henriet P, Jodele S, Emonard H, Feron O, Courtoy PJ, DeClerck YA, Marbaix E: Mechanisms of pericyte recruitment in tumour angiogenesis: a new role for metalloproteinases. Eur J Cancer 2006, 42(3):310-318.
• [9]Shimoda M, Mellody KT, Orimo A: Carcinoma-associated fibroblasts are a rate-limiting determinant for tumour progression. Semin Cell Dev Biol 2010, 21(1):19-25.
• [10]Mantoni TS, Lunardi S, Al-Assar O, Masamune A, Brunner TB: Pancreatic stellate cells radioprotect pancreatic cancer cells through beta1-integrin signaling. Cancer Res 2011, 71(10):3453-3458.
• [11]Hawsawi NM, Ghebeh H, Hendrayani SF, Tulbah A, Al-Eid M, Al-Tweigeri T, Ajarim D, Alaiya A, Dermime S, Aboussekhra A: Breast carcinoma-associated fibroblasts and their counterparts display neoplastic-specific changes. Cancer Res 2008, 68(8):2717-2725.
• [12]Hwang RF, Moore T, Arumugam T, Ramachandran V, Amos KD, Rivera A, Ji B, Evans DB, Logsdon CD: Cancer-associated stromal fibroblasts promote pancreatic tumor progression. Cancer Res 2008, 68(3):918-926.
• [13]Rodier F, Coppé JP, Patil CK, Hoeijmakers WA, Muñoz DP, Raza SR, Freund A, Campeau E, Davalos AR, Campisi J: Persistent DNA damage signalling triggers senescence-associated inflammatory cytokine secretion. Nat Cell Biol 2009, 11(8):973-979.
• [14]Ohuchida K, Mizumoto K, Murakami M, Qian LW, Sato N, Nagai E, Matsumoto K, Nakamura T, Tanaka M: Radiation to stromal fibroblasts increases invasiveness of pancreatic cancer cells through tumor-stromal interactions. Cancer Res 2004, 64(9):3215-3222.
• [15]Papadopoulou A, Kletsas D: Human lung fibroblasts prematurely senescent after exposure to ionizing radiation enhance the growth of malignant lung epithelial cells in vitro and in vivo. Int J Oncol 2011, 39(4):989-999.
• [16]Garcia-Barros M, Paris F, Cordon-Cardo C, Lyden D, Rafii S, Haimovitz-Friedman A, Fuks Z, Kolesnick R: Tumor response to radiotherapy regulated by endothelial cell apoptosis. Science 2003, 300(5622):1155-1159.
• [17]Lee Y, Wang Y, Burnette B, Wang Y, Meng Y, Beckett M, Sharma R, Chin R, Tu T, Weichselbaum RR, Fu YX: Therapeutic effects of ablative radiation on local tumor require CD8+ T cells: changing strategies for cancer treatment. Blood 2009, 114(3):589-595.
• [18]Brown JM, Koong AC: High-dose single-fraction radiotherapy: exploiting a new biology? Int J Radiat Oncol Biol Phys 2008, 71(2):324-325.
• [19]Xing JZ, Zhu L, Gabos S, Xie L: Microelectronic cell sensor assay for detection of cytotoxicity and prediction of acute toxicity. Toxicol In Vitro 2006, 20(6):995-1004.
• [20]Atienza JM, Yu N, Kirstein SL, Xi B, Wang X, Xu X, Abassi YA: Dynamic and label-free cell-based assays using the real-time cell electronic sensing system. Assay Drug Dev Technol 2006, 4(5):597-607.
• [21]Abassi YA, Xi B, Zhang W, Ye P, Kirstein SL, Gaylord MR, Feinstein SC, Wang X, Xu X: Kinetic cell-based morphological screening: prediction of mechanism of compound action and off-target effects. Chem Biol 2009, 16(7):712-723.
• [22]Eisenberg MC, Kim Y, Li R, Ackerman WE, Kniss DA, Friedman A: Mechanistic modeling of the effects of myoferlin on tumor cell invasion. Proc Natl Acad Sci USA 2011, 108(50):20078-20083.
• [23]Scanlan MJ, Raj BKM, Calvo B, Garin-Chesa P, Sanz-Moncasi MP, Healey JH, Old LJ, Rettig WJ: Molecular cloning of fibroblast activation protein alpha, a member of the serine protease family selectively expressed in stromal fibroblasts of epithelial cancers. Proc Natl Acad Sci USA 1994, 91(12):5657-5661.
• [24]Park JE, Lenter MC, Zimmermann RN, Garin-Chesa P, Old LJ, Rettig WJ: Fibroblast activation protein, a dual specificity serine protease expressed in reactive human tumor stromal fibroblasts. J Biol Chem 1999, 274(51):36505-36512.
• [25]d'Adda di Fagagna F, Reaper PM, Clay-Farrace L, Fiegler H, Carr P, von Zglinicki T, Saretzki G, Carter NP, Jackson SP: A DNA damage checkpoint response in telomere-initiated senescence. Nature 2003, 426(6963):194-198.
• [26]Efimova EV, Mauceri HJ, Golden DW, Labay E, Bindokas VP, Darga TE, Chakraborty C, Barreto-Andrade JC, Crawley C, Sutton HG, Kron SJ, Weichselbaum RR: Poly(ADP-ribose) polymerase inhibitor induces accelerated senescence in irradiated breast cancer cells and tumors. Cancer Res 2010, 70(15):6277-6282.
• [27]McCarthy JB, Basara ML, Palm SL, Sas DF, Furcht LT: The role of cell adhesion proteins-laminin and fibronectin-in the movement of malignant and metastatic cells. Cancer Metastasis Rev 1985, 4(2):125-152.
• [28]Suzuki M, Boothman DA: Stress-induced premature senescence (SIPS)-influence of SIPS on radiotherapy. J Radiat Res (Tokyo) 2008, 49(2):105-112.
• [29]Wang D, Jang DJ: Protein kinase CK2 regulates cytoskeletal reorganization during ionizing radiation-induced senescence of human mesenchymal stem cells. Cancer Res 2009, 69(20):8200-8207.
• [30]Banath JP, Klokov D, MacPhail SH, Banuelos CA, Olive PL: Residual gammaH2AX foci as an indication of lethal DNA lesions. BMC Cancer 2010, 10:4. BioMed Central Full Text
• [31]Coussens LM, Tinkle CL, Hanahan D, Werb Z: MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis. Cell 2000, 103(3):481-490.
• [32]Murray GI, Duncan ME, O'Neil P, Melvin WT, Fothergill JE: Matrix metalloproteinase-1 is associated with poor prognosis in colorectal cancer. Nat Med 1996, 2(4):461-462.
• [33]Holleran WM, Galardy RE, Gao WN, Levy D, Tang PC, Elias PM: Matrix metalloproteinase inhibitors reduce phorbol ester-induced cutaneous inflammation and hyperplasia. Arch Dermatol Res 1997, 289(3):138-144.
• [34]Sternlicht MD, Lochter A, Sympson CJ, Huey B, Rougier JP, Gray JW, Pinkel D, Bissell MJ, Werb Z: The stromal proteinase MMP3/stromelysin-1 promotes mammary carcinogenesis. Cell 1999, 98(2):137-146.
• [35]Martin MD, Matrisian LM: The other side of MMPs: protective roles in tumor progression. Cancer Metastasis Rev 2007, 26(3-4):717-724.
• [36]Kessenbrock K, Plaks V, Werb Z: Matrix metalloproteinases: regulators of the tumor microenvironment. Cell 2010, 141(1):52-67.
• [37]Desgrosellier JS, Cheresh DA: Integrins in cancer: biological implications and therapeutic opportunities. Nat Rev Cancer 2010, 10(1):9-22.
• [38]Hynes RO: Integrins: bidirectional, allosteric signaling machines. Cell 2002, 110(6):673-687.
• [39]Caswell PT, Vadrevu S, Norman JC: Integrins: masters and slaves of endocytic transport. Nat Rev Mol Cell Biol 2009, 10(12):843-853.
• [40]Teckchandani A, Toida N, Goodchild J, Henderson C, Watts J, Wollscheid B, Cooper JA: Quantitative proteomics identifies a Dab2/integrin module regulating cell migration. J Cell Biol 2009, 186(1):99-111.
• [41]Friedl P, Wolf K: Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 2003, 3(5):362-374.
• [42]Cordes N, Seidler J, Durzok R, Geinitz H, Brakebusch C: beta1-integrin-mediated signaling essentially contributes to cell survival after radiation-induced genotoxic injury. Oncogene 2006, 25(9):1378-1390.
• [43]Zhang Z, Vuori K, Reed JC, Ruoslahti E: The alpha 5 beta 1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression. Proc Natl Acad Sci USA 1995, 92(13):6161-6165.
• [44]Park CC, Zhang HJ, Yao ES, Park CJ, Bissell MJ: Beta1 integrin inhibition dramatically enhances radiotherapy efficacy in human breast cancer xenografts. Cancer Res 2008, 68(11):4398-4405.