Department of Radiation Oncology (Chen BY,Gu HM,Zhou SZ,Yuan JP,Qain X), Department of Gastroenterology (Li XY,Xiao MB,Jiang F),Affiliated Hospital of Nantong University,Nantong 226001,China
Abstract： Objective To investigate the effects of high-energy X-ray irradiation on collagen expression in human fetal lung fibroblasts (HFL-1) and identify the genes related to collagen production induced by X-ray, and to provide a theoretical basis for the clinical treatment of radiation-induced pulmonary fibrosis. Methods HFL-1 cells cultured in vitro were randomly divided into control group and irradiation group. The irradiation group was irradiated with 6 MV X-ray at a single dose of 5 Gy. At 24 hours after irradiation, the expression of hydroxyproline (HYP) in the two groups was determined by the digestion method. The expression of collagen types Ⅰ and Ⅲ at mRNA and protein levels was measured by RT-PCR and Western blot. And gene chip was used to identify the differentially expressed genes between the two groups. Results At 24 hours after irradiation, the irradiation group had significantly higher expression of HYP than the control group (1.834 μg/ml vs.4.912 μg/ml, P=0.000), and the irradiation group had significantly higher mRNA and protein expression of collagen types Ⅰ and Ⅲ than the control group (18.535∶vs. 86.779, P=0.000;4.337∶vs. 4.425, P=0.000). A total of 1879 genes were differentially expressed between the two groups, including 771 up-regulated genes and 1108 down-regulated genes in the irradiation group. The genes involved in fibrosis which were up-regulated more than 5-fold were transforming growth factor (TGF)-beta 1, TGF-beta 3, matrix metalloproteinase (MMP)-28, MMP-26, MMP-27, and SMAD6. Conclusions High-energy X-ray can induce collagen production in HFL-1 cells. Gene chip can effectively identify the genes related to collagen production induced by X-ray. Many different genes areinvolved in the development and progression of radiation-induced pulmonary fibrosis.
Qian Xia,Li Xiaoyan,Xiao Mingbing et al. Application of gene chip in identifying genes related to collagen production induced by X-ray in HFL-1 cells[J]. Chinese Journal of Radiation Oncology, 2017, 26(3): 329-334.
[1] Ding NH,Li JJ,Sun LQ.Molecular mechanisms and treatment of radiation-induced lung fibrosis[J].Curr Drug Targets,2013,14(11):1347-1356.DOI:10.2174/13894501113149990198. [2] Marks LB,Bentzen SM,Deasy JO,et al. Radiation dose-volume effects in the lung[J].Int J Radiat Oncol Biol Phys,2010,76(3 Suppl):S70-76.DOI:10.1016/j.ijrobp.2009.06.091 [3] Yarnold J,Brotons MCV.Pathogenetic mechanisms in radiation fibrosis[J].Radiother Oncol,2010,97(1):149-161.DOI:10.1016/j.radonc.2010.09.002. [4] Sakai N,Tager AM.Fibrosis of two:epithelial cell-fibroblast interactions in pulmonary fibrosis[J].Biochim Biophys Acta,2013,1832(7):911-921.DOI:10.1016/j.bbadis.2013.03.001. [5] 陈不尤,李曙光,肖明兵,等.放射性肺炎方抗大鼠放射性肺纤维化作用机制的研究[J].中华放射医学与防护杂志,2012,32(5):475-480.DOI:10.3760/cma.j.issn.0254-5098.2012.05.006. Chen BY,Li SG,Xiao MB,et al. The mechanisms of ARPD in treating radiation-induced lung fibrosis in rats[J].Chin J Radiol Med Prot,2012,32(5):475-480.DOI:10.3760/cma.j.issn.0254-5098.2012.05.006. [6] Van Hoozen BE,Grimmer KL,Marelich GP,et al. Early phase collagen synthesis in lungs of rats exposed to bleomycin[J].Toxicology,2000,147(1):1-13.DOI:10.1016/S0300-483X (00)00142-6. [7] Trost B,Moir CA,Gillespie ZE,et al. Concordance between RNA-sequencing data and DNA microarray data in transcriptome analysis of proliferative and quiescent fibroblasts[J].R Soc Open Sci,2015,2(9):150402.DOI:10.1098/rsos.150402. [8] Ciccia F,Guggino G,Rizzo A,et al. Interleukin (IL)-22 receptor 1 is over-expressed in primary Sjogren′s syndrome and Sjgren-associated non-Hodgkin lymphomas and is regulated by IL-18[J].Clin Exp Immunol,2015,181(2):219-229.DOI:10.1111/cei.12643. [9] Song ZW.Roles of the nucleotide sugar transporters (SLC35 family) in health and disease[J].Mol Aspects Med,2013,34(2-3):590-600.DOI:10.1016/j.mam.2012.12.004. [10] Ladd AN,Charlet-B N,Cooper TA.The CELF family of RNA binding proteins is implicated in cell-specific and developmentally regulated alternative splicing[J].Mol Cell Biol,2001,21(4):1285-1296.DOI:10.1128/MCB.21.4.1285-1296.2001. [11] Xiao J,Ho CT,Liong EC,et al. Epigallocatechin gallate attenuates fibrosis,oxidative stress,and inflammation in non-alcoholic fatty liver disease rat model through TGF/SMAD,PI3 K/Akt/FoxO1,and NF-kappa B pathways[J].Eur J Nutr,2014,53(1):187-199.DOI:10.1007/s00394-013-0516-8. [12] Yao Q,Pawlaczyk K,Ayala ER,et al. The role of the TGF/Smad signaling pathway in peritoneal fibrosis induced by peritoneal dialysis solutions[J].Nephron Exp Nephrol,2008,109(2):e71-78.DOI:10.1159/000142529. [13] Chang KC,Petrash JM.Aldose reductase mediates transforming growth factor β2(TGF-β2)-induced migration and epithelial-to-mesenchymal transition of lens-derived epithelial cells[J].Invest Ophthalmol Vis Sci,2015,56(8):4198-4210.DOI:10.1167/iovs.15-16557. [14] Edwards DR,Murphy G,Reynolds JJ,et al. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor[J].EMBO J,1987,6(7):1899-1904. [15] Rubin P,Johnston CJ,Williams JP,et al. A perpetual cascade of cytokines postirradiation leads to pulmonary fibrosis[J].Int J Radiat Oncol Biol Phys,1995,33(1):99-109.DOI:10.1016/0360-3016(95)00095-G. [16] Nayak RR,Kearns M,Spielman RS,et al. Coexpression network based on natural variation in human gene expression reveals gene interactions and functions[J].Genome Res,2009,19(11):1953-1962.DOI:10.1101/gr.097600.109.
2017, Vol. 26 
