Abstract
Radiotherapy is a major method for cancer treatment, but it frequently causes various side effects such as radiation-induced pneumonia and pulmonary fibrosis. Drugs to treat these side effects are urgently needed in the clinic, since there is no clearly defined medication for treating these symptoms. Previous studies demonstrated that the herbal formula, PM014, is effective for radiation-induced lung injury and fibrosis in mice. In this study, we investigated gene expression profiles to understand the mechanism of action behind the effects of PM014 on radiation induced damage in immortalized lung epithelial cells, MLE12. We performed QuantSeq 3′ mRNA-Seq analysis on the mRNA from radiation treated MLE-12 cells in the presence and absence of PM014. Transcriptome analysis found that 217 genes were significantly affected by PM014. Among them (217 genes, > twofold, p value < 0.05, 4 normalize), 77 genes were found to be upregulated, and 140 genes were downregulated in response to PM014 treatment in a dose dependent manner. Using the Kyoto Encyclopedia of Genes and Genomes analysis, we found that genes involved in cytokine–cytokine receptor interaction pathways were the most strongly affected by PM014. Based on these data, we selected 20 genes, and performed real-time PCR. Expression of 11 genes, including IL-18, IL-12a, Tnfrsf9, IL-17, CCR5, Blnk, Irf8, Nrros, TGF-β, Relt, and Cxcl2 was increased after irradiation, while PM014 treatment showed the reversed expression pattern of these genes. Therefore, PM014 may be useful for the treatment of radiation induced lung injury by modulating genes involved in cytokine–cytokine receptor interaction pathway.
Similar content being viewed by others
References
Agostini C, Gurrieri C (2006) Chemokine/cytokine cocktail in idiopathic pulmonary fibrosis. Proc Am Thorac Soc 3:357–363. https://doi.org/10.1513/pats.200601-010TK
Borthwick LA, Wynn TA, Fisher AJ (2013) Cytokine mediated tissue fibrosis. Biochem Biophys Acta 1832:1049–1060. https://doi.org/10.1016/j.bbadis.2012.09.014
Branton MH, Kopp JB (1999) TGF-beta and fibrosis. Microbes Infect 1:1349–1365
Castranova V, Rabovsky J, Tucker JH, Miles PR (1988) The alveolar type II epithelial cell: a multifunctional pneumocyte. Toxicol Appl Pharmacol 93:472–483
Chiang CH, Chuang CH, Liu SL (2014) Transforming growth factor-beta1 and tumor necrosis factor-alpha are associated with clinical severity and airflow limitation of COPD in an additive manner. Lung 192:95–102. https://doi.org/10.1007/s00408-013-9520-2
Gentleman RC et al (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome biol 5:R80. https://doi.org/10.1186/gb-2004-5-10-r80
Jordan JA et al (2001) Role of IL-18 in acute lung inflammation. J Immunol 167:7060–7068
Jung KH et al (2014) The effects of the standardized herbal formula PM014 on pulmonary inflammation and airway responsiveness in a murine model of cockroach allergen-induced asthma. J Ethnopharmacol 155:113–122. https://doi.org/10.1016/j.jep.2014.04.029
Kapanci Y, Desmouliere A, Pache JC, Redard M, Gabbiani G (1995) Cytoskeletal protein modulation in pulmonary alveolar myofibroblasts during idiopathic pulmonary fibrosis. Possible role of transforming growth factor beta and tumor necrosis factor alpha. Am J Respir Crit Care Med 152:2163–2169. https://doi.org/10.1164/ajrccm.152.6.8520791
Kim JY et al (2017) Standardized herbal formula PM014 inhibits radiation-induced pulmonary inflammation in mice. Sci Rep 7:45001. https://doi.org/10.1038/srep45001
Langmead B, Salzberg SL (2012) Fast gapped-read alignment with Bowtie 2. Nat methods 9:357–359. https://doi.org/10.1038/nmeth.1923
Lee H et al (2012) Herbal formula, PM014, attenuates lung inflammation in a murine model of chronic obstructive pulmonary disease. Evid Based Complement Altern Med 2012:769830. https://doi.org/10.1155/2012/769830
Marks LB et al (2010) Radiation dose-volume effects in the lung. Int J Radiat Oncol Biol Phys 76:S70–S76. https://doi.org/10.1016/j.ijrobp.2009.06.091
Nguefack-Tsague G, Zucchini W, Fotso S (2016) Frequentist model averaging and applications to bernoulli trials. Open J Stat 6:545–553
Okamura H et al (1995) Cloning of a new cytokine that induces IFN-gamma production by T cells. Nature 378:88–91. https://doi.org/10.1038/378088a0
Puchelle E, Zahm JM, Tournier JM, Coraux C (2006) Airway epithelial repair, regeneration, and remodeling after injury in chronic obstructive pulmonary disease. Proc Am Thorac Soc 3:726–733. https://doi.org/10.1513/pats.200605-126sf
Quinlan AR, Hall IM (2010) BEDTools: a flexible suite of utilities for comparing genomic features. Bioinform 26:841–842. https://doi.org/10.1093/bioinformatics/btq033
Raman K, Kaplan MH, Hogaboam CM, Berlin A, Lukacs NW (2003) STAT4 signal pathways regulate inflammation and airway physiology changes in allergic airway inflammation locally via alteration of chemokines. J Immunol 170:3859–3865
Schindler H, Lutz MB, Rollinghoff M, Bogdan C (2001) The production of IFN-gamma by IL-12/IL-18-activated macrophages requires STAT4 signaling and is inhibited by IL-4. J Immunol 166:3075–3082
Sime PJ, O’Reilly KM (2001) Fibrosis of the lung and other tissues: new concepts in pathogenesis and treatment. Clin Immunol 99:308–319. https://doi.org/10.1006/clim.2001.5008
Sun F, Xiao G, Qu Z (2017) Isolation of murine alveolar type II epithelial cells. Bio Protoc. https://doi.org/10.21769/bioprotoc.2288
Trinchieri G, Gerosa F (1996) Immunoregulation by interleukin-12. J Leukoc Biol 59:505–511
Wu Z et al (2013) Effects of carbon ion beam irradiation on lung injury and pulmonary fibrosis in mice. Exp Ther Med 5:771–776. https://doi.org/10.3892/etm.2013.881
Yamauchi Y et al (2010) Tumor necrosis factor-alpha enhances both epithelial-mesenchymal transition and cell contraction induced in A549 human alveolar epithelial cells by transforming growth factor-beta1. Exp Lung Res 36:12–24. https://doi.org/10.3109/01902140903042589
Yoo H, Kang JW, Lee DW, Oh SH, Lee YS, Lee EJ, Cho J (2015) Pyruvate metabolism: a therapeutic opportunity in radiation-induced skin injury. Biochem Biophys Res Commun 460:504–510. https://doi.org/10.1016/j.bbrc.2015.03.060
Acknowledgements
This work was supported by the Convergence of Conventional Medicine and Traditional Korean Medicine R&D program funded by the Ministry of Health & Welfare through the Korea Health Industry Development Institute (HI15C0214).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Ethical statement
N/A.
Conflict of interest
The authors have no conflict of interest to declare.
Rights and permissions
About this article
Cite this article
Shin, D., Joo, K.G., Kang, M.J. et al. Gene expression profile of PM014 of immortalized mouse lung epithelial cells in response to the effect of PM014 on radiation-induced fibrosis. Orient Pharm Exp Med 19, 107–114 (2019). https://doi.org/10.1007/s13596-018-0350-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13596-018-0350-x