Patterns of protein expression in human head and neck cancer cell lines differ after proton vs photon radiotherapy
Li Wang MD, PhD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorLiuqing Yang PhD
Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorShichao Han MD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorJinming Zhu MD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorYuting Li PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorZeming Wang BS
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorYou-Hong Fan BS
Department of Thoracic/Head & Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorEric Lin MS
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorRuiping Zhang MS
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorNarayan Sahoo PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorYupeng Li MS
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorXiaodong Zhang PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorXiaochun Wang PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorTengfei Li PhD
Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorXiaorong R. Zhu PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorHongtu Zhu PhD
Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorJohn V. Heymach MD, PhD
Department of Thoracic/Head & Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorJeffrey N. Myers MD, PhD
Department of Head & Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorCorresponding Author
Steven J. Frank MD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Correspondence
Steven J. Frank, MD, Department of Radiation Oncology, Unit 1150, The University of Texas MD Anderson Cancer Center, 1840 Old Spanish Trail, Houston, TX 77054-1901.
Email: [email protected]
Search for more papers by this authorLi Wang MD, PhD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorLiuqing Yang PhD
Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorShichao Han MD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorJinming Zhu MD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorYuting Li PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorZeming Wang BS
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorYou-Hong Fan BS
Department of Thoracic/Head & Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorEric Lin MS
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorRuiping Zhang MS
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorNarayan Sahoo PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorYupeng Li MS
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorXiaodong Zhang PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorXiaochun Wang PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorTengfei Li PhD
Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorXiaorong R. Zhu PhD
Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorHongtu Zhu PhD
Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorJohn V. Heymach MD, PhD
Department of Thoracic/Head & Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorJeffrey N. Myers MD, PhD
Department of Head & Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, Texas
Search for more papers by this authorCorresponding Author
Steven J. Frank MD
Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas
Correspondence
Steven J. Frank, MD, Department of Radiation Oncology, Unit 1150, The University of Texas MD Anderson Cancer Center, 1840 Old Spanish Trail, Houston, TX 77054-1901.
Email: [email protected]
Search for more papers by this authorFunding information: The University of Texas MD Anderson Cancer Center; Institutional Research Grant program; Sister Institution Network Fund; Cancer Center Support (Core) Grant, Grant/Award Number: P30CA016672
Abstract
Background
Proton radiotherapy (PRT) may be a less toxic alternative to photon radiotherapy (XRT) for patients with head and neck squamous cell carcinoma (HNSCC). However, the molecular responses of HNSCC cells to PRT vs XRT are unclear.
Methods
Proteomics analyses of protein expression profiles by reverse-phase protein arrays were done for two human papillomavirus [HPV]-negative and two HPV+ cell lines. Expression patterns of 175 proteins involved in several signaling pathways were tested.
Results
Compared with PRT, XRT tended to induce lower expression of DNA damage repair—and cell cycle arrest-related proteins and higher expression of cell survival- and proliferation-related proteins.
Conclusions
Under these experimental conditions, PRT and XRT induced different protein expression and activation profiles. Further preclinical verification is needed, as are studies of tumor pathway mutations as biomarkers for choice of treatment or as radiosensitization targets to improve the response of HNSCC to PRT or XRT.
CONFLICTS OF INTEREST
Steven J. Frank is a scientific advisory board member of Breakthrough Chronic Care; he has received research grants from C4 Imaging, Eli Lilly, Elekta, and Hitachi; and he has reported personal fees from Varian Medical Systems, Inc. (Consultant/Advisory Board), C4 Imaging (Founder and Director), Hitachi (Honoraria/Advisory Board), Augmenix (Honoraria), and National Comprehensive Cancer Center (Board member). John V. Heymach is advisory committees of AstraZeneca, BioTree, Boehringer Ingelheim, Exelixis, Genentech, GlaxoSmithKline, Guardant Health, Hengrui Medicine, Eli Lilly, Novartis, Spectrum Pharmaceuticals, Inc., EMD Serono, and Synta Pharmaceuticals; he has received research support from AstraZeneca, Bayer, GlaxoSmithKline and Spectrum Pharmaceuticals, Inc.; he also has received royalties and licensing fees from Spectrum Pharmaceuticals, Inc. and BioTree. Other authors declare no conflicts of interest regarding the content of this article.
Supporting Information
| Filename | Description |
|---|---|
| hed26005-sup-0001-FigureS1.TIFTIFF image, 2 MB | Fig. S1 Cell radiation setting. |
| hed26005-sup-0002-FigureS2.TIFTIFF image, 4.6 MB | Fig. S2 The protein expression profile dynamic changes in four head and neck squamous cell carcinoma cell lines after photon (XRT) vs proton (PRT) irradiation in heat map of Figure 2A re-mapped by a similar group of genes across all three time points and a time course for genes that are differently regulated across all three time points. |
| hed26005-sup-0003-TableS1.xlsxExcel 2007 spreadsheet , 22.5 KB | Table S1 Antibosy list of the Reverse-Phase Protein Array |
| hed26005-sup-0004-TableS2.xlsxExcel 2007 spreadsheet , 13.7 KB | Table S2 Protein expression profile differences in HNSCC cells treated with X-ray vs Proton |
| hed26005-sup-0005-TableS3.xlsxExcel 2007 spreadsheet , 23.4 KB | Table S3 The protein expression profile dynamic changes of HNSCC cells treated with X-ray vs Proton |
| hed26005-sup-0006-TableS4.xlsxExcel 2007 spreadsheet , 315 KB | Table S4 Protein expression relative value of proton or X-ray to no treatment cells |
| hed26005-sup-0007-TableS5.xlsxExcel 2007 spreadsheet , 15.1 KB | Table S5 Protein expression profile in HNSCC cells with different HPV status treated with X-rays vs protons |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin. 2010; 60: 277-300.
- 2Setton J, Lee NY, Riaz N, et al. A multi-institution pooled analysis of gastrotomy tube dependence in patients with oropharyngeal cancer treated with definitive intensity-modulated radiotherapy. Cancer. 2015; 121(2): 294-301.
- 3Argiris A, Brockstein BE, Haraf DJ, et al. Competing causes of death and second primary tumors in patients with locoregionally advanced head and neck cancer treated with chemoradiotherapy. Clin Cancer Res. 2004; 10(6): 1956-1962.
- 4Mell LK, Dignam JJ, Salama JK, et al. Predictors of competing mortality in advanced head and neck cancer. J Clin Oncol. 2010; 28: 15-20.
- 5Echarri MJ, Lopez-Martin A, Hitt R. Targeted therapy in locally advanced and recurrent/metastatic head and neck squamous cell carcinoma. Cancers (Basel). 2016; 8(3): 27.
- 6Ausoni S, Boscolo-Rizzo P, Singh B, et al. Targeting cellular and molecular drivers of head and neck squamous cell carcinoma: current options and emerging perspectives. Cancer Metastasis Rev. 2016; 35: 413-426.
- 7Durante M, Loeffler JS. Charged particles in radiation oncology. Nat Rev Clin Oncol. 2010; 7: 37-43.
- 8Frese MC, Yu VK, Stewart RD, Carlson DJ. A mechanism-based approach to predict the relative biological effectiveness of protons and carbon ions in radiation therapy. Int J Radiat Oncol Biol Phys. 2012; 83: 442-450.
- 9Blanchard P, Garden AS, Gunn GB, et al. Intensity-modulated proton beam therapy (IMPT) versus intensity-modulated photon therapy (IMRT) for patients with oropharynx cancer - a case matched analysis. Radiother Oncol. 2016; 120(1): 48-55.
- 10Romesser PB, Cahlon O, Scher E, et al. Proton beam radiation therapy results in significantly reduced toxicity compared with intensity-modulated radiation therapy for head and neck tumors that require ipsilateral radiation. Radiother Oncol. 2016; 118(2): 286-292.
- 11Gridley DS, Pecaut MJ, Mao XW, Wroe AJ, Luo-Owen X. Biological effects of passive versus active scanning proton beams on human lung epithelial cells. Technol Cancer Res Treat. 2015; 14(1): 81-98.
- 12Kwon YS, Lee KS, Chun SY, Jang TJ, Nam KS. Suppressive effects of a proton beam on tumor growth and lung metastasis through the inhibition of metastatic gene expression in 4T1 orthotopic breast cancer model. Int J Oncol. 2016; 49(1): 336-342.
- 13Ha BG, Jung SS, Shon YH. Effects of proton beam irradiation on mitochondrial biogenesis in a human colorectal adenocarcinoma cell line. Int J Oncol. 2017; 51(3): 859-866.
- 14Ha BG, Park JE, Cho HJ, Lim YB, Shon YH. Inhibitory effects of proton beam irradiation on integrin expression and signaling pathway in human colon carcinoma HT29 cells. Int J Oncol. 2015; 46(6): 2621-2628.
- 15Beheshti A, Peluso M, Lamont C, Hahnfeldt P, Hlatky L. Proton irradiation augments the suppression of tumor progression observed with advanced age. Radiat Res. 2014; 181(3): 272-283.
- 16Finnberg N, Wambi C, Ware JH, Kennedy AR, El-Deiry WS. Gamma-radiation (GR) triggers a unique gene expression profile associated with cell death compared to proton radiation (PR) in mice in vivo. Cancer Biol Ther. 2008; 7(12): 2023-2033.
- 17Nielsen S, Bassler N, Grzanka L, et al. Differential gene expression in primary fibroblasts induced by proton and cobalt-60 beam irradiation. Acta Oncol. 2017; 56(11): 1406-1412.
- 18Girdhani S, Lamont C, Hahnfeldt P, Abdollahi A, Hlatky L. Proton irradiation suppresses angiogenic genes and impairs cell invasion and tumor growth. Radiat Res. 2012; 178(1): 33-45.
- 19Narang H, Kumar A, Bhat N, Pandey BN, Ghosh A. Effect of proton and gamma irradiation on human lung carcinoma cells: gene expression, cell cycle, cell death, epithelial-mesenchymal transition and cancer-stem cell trait as biological end points. Mutat Res. 2015; 780: 35-46.
- 20Bravatà V, Minafra L, Cammarata FP, et al. Gene expression profiling of breast cancer cell lines treated with proton and electron radiations. Br J Radiol. 2018; 91(1089): 20170934.
- 21Winter M, Dokic I, Schlegel J, et al. Deciphering the acute cellular phosphoproteome response to irradiation with X-rays, protons and carbon ions. Mol Cell Proteomics. 2017; 16(5): 855-872.
- 22Bedford JS. Sublethal damage, potentially lethal damage, and chromosomal aberrations in mammalian cells exposed to ionizing radiations. Int J Radiat Oncol Biol Phys. 1991; 21(6): 1457-1469.
- 23Wang L, Wang X, Li Y, et al. Human papillomavirus status and the relative biological effectiveness of protons radiotherapy in head and neck cancer cells. Head Neck. 2017; 39(4): 708-715.
- 24Wang L, Han S, Zhu J, et al. Proton versus photon radiation-induced cell death in head and neck cancer cells. Head Neck. 2019; 41(1): 46-55.
- 25Green LM, Murray DK, Bant AM, et al. Response of thyroid follicular cells to gamma irradiation compared to proton irradiation. I. Initial characterization of DNA damage, micronucleus formation, apoptosis, cell survival, and cell cycle phase redistribution. Radiat Res. 2001; 155: 32-42.
- 26Paganetti H, Blakely E, Carabe-Fernandez A, et al. Report of the AAPM TG-256 on the relative biological effectiveness of proton beams in radiation therapy. Med Phys. 2019; 46(3): e53-e78.
- 27Chung CH, Gillison ML. Human papillomavirus in head and neck cancer: its role in pathogenesis and clinical implications. Clin Cancer Res. 2009; 15: 6758-6762.
- 28Ang KK, Harris J, Wheeler R, et al. Human papillomavirus and survival of patients with oropharyngeal cancer. N Engl J Med. 2010; 363(1): 24-35.
- 29Andreo P, Burns DT, Hohlfield K, et al. Absorbed dose determination in external beam radiotherapy: an international code of practice for dosimetry based on standards of absorbed dose to water. IAEA TRS-398 V12 International Atomic Energy Agency, IAEA in Austria, June 5, 2006 (V.12).
- 30Almond PR, Biggs PJ, Coursey BM, et al. AAPM's TG 51 protocol for clinical reference dosimetry of high-energy photon and electron beams. Med Phys. 1999; 26(9): 1847-1870.
- 31Mazumdar T, Byers LA, Ng PK, et al. A comprehensive evaluation of biomarkers predictive of response to PI3K inhibitors and of resistance mechanisms in head and neck squamous cell carcinoma. Mol Cancer Ther. 2014; 13(11): 2738-2750.
- 32Byers LA, Wang J, Nilsson MB, et al. Proteomic profiling identifies dysregulated pathways in small cell lung cancer and novel therapeutic targets including PARP1. Cancer Discov. 2012; 2(9): 798-811.
- 33Pin E, Stratton S, Belluco C, et al. A pilot study exploring the molecular architecture of the tumor microenvironment in human prostate cancer using laser capture microdissection and reverse phase protein microarray. Mol Oncol. 2016; 10(10): 1585-1594.
- 34Datta J, Damodaran S, Parks H, et al. Akt activation mediates acquired resistance to fibroblast growth factor receptor inhibitor BGJ398. Mol Cancer Ther. 2017; 16(4): 614-624.
- 35Storey JD, Tibshirani R. Statistical significance for genomewide studies. Proc Natl Acad Sci U S A. 2003; 100(16): 9440-9445.
- 36Mao C, Howard TD, Sullivan D, et al. Bioinformatic analysis of coronary disease associated SNPs and genes to identify proteins potentially involved in the pathogenesis of atherosclerosis. J Proteom Genom Res. 2017; 2(1): 1-12.
- 37Dyshlovoy SA, Otte K, Venz S, et al. Proteomic-based investigations on the mode of action of the marine anticancer compound rhizochalinin. Proteomics. 2017; 17(11): 1700048.
- 38Grosse N, Fontana AO, Hug EB, et al. Deficiency in homologous recombination renders mammalian cells more sensitive to proton versus photon irradiation. Int J Radiat Oncol Biol Phys. 2014; 88(1): 175-181.
- 39Cartwright IM, Bell JJ, Maeda J, et al. Effects of targeted phosphorylation site mutations in the DNA-PKcs phosphorylation domain on low and high LET radiation sensitivity. Oncol Lett. 2015; 9(4): 1621-1627.
- 40Glorieux M, Dok R, Nuyts S. Novel DNA targeted therapies for head and neck cancers: clinical potential and biomarkers. Oncotarget. 2017; 8(46): 81662-81678.
- 41Berger FG, Berger SH. Thymidylate synthase as a chemotherapeutic drug target: where are we after fifty years? Cancer Biol Ther. 2006; 5(9): 1238-1241.
- 42Diegeler S, Hellweg CE. Intercellular communication of tumor cells and immune cells after exposure to different ionizing radiation qualities. Front Immunol. 2017; 8: 664.
- 43Twyman-Saint Victor C, Rech AJ, Maity A, et al. Radiation and dual checkpoint blockade activate non-redundant immune mechanisms in cancer. Nature. 2015; 520(7547): 373-377.
- 44Ebner DK, Tinganelli W, Helm A, et al. The immunoregulatory potential of particle radiation in cancer therapy. Front Immunol. 2017; 8: 99.
- 45Fernandez-Gonzalo R, Baatout S, Moreels M. Impact of particle irradiation on the immune system: from the clinic to Mars. Front Immunol. 2017; 8: 177.
- 46Cullen SP, Brunet M, Martin SJ. Granzymes in cancer and immunity. Cell Death Differ. 2010; 17(4): 616-623.
- 47Hermiston ML, Xu Z, Weiss A. CD45: a critical regulator of signaling thresholds in immune cells. Annu Rev Immunol. 2003; 21: 107-137.
- 48Venkataramani V, Küffer S, Cheung KCP, et al. CD31 expression determines redox status and chemoresistance in human angiosarcomas. Clin Cancer Res. 2018; 24(2): 460-473.
- 49Bristow RG, Alexander B, Baumann M, et al. Combining precision radiotherapy with molecular targeting and immunomodulatory agents: a guideline by the American Society for Radiation Oncology. Lancet Oncol. 2018; 19(5): e240-e251.
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