Abstract：Objective To establish radiation-resistant lung carcinoma cell lines, and to investigate the changes in morphology, apoptosis, invasive migration, and epithelial-mesenchymal transition (EMT) in cells. Methods The radiation-resistant lung carcinoma cell lines were obtained by exposure of lung carcinoma cell lines, A549 and H1299, to radiation with a low dose in fractions, a sublethal dose, or a gradually increasing dose. The morphological changes in cells, radiosensitivity, survival rates after exposure, apoptosis rates, changes in invasive migration, and expression of EMT marker proteins were evaluated using microscopy, colony formation assay, CCK-8 assay, flow cytometry, transwell migration assay, and Western blot, respectively. Results Radiation with a gradually increasing dose successfully induced the radiation-resistant cell lines, A549R and H1299R. The morphological study showed that the morphology of radiation-resistant cells was converted to the morphology of mesenchymal cells. Compared with A549 and H1299 cells, the values of D0, Dq, and SF2 were significantly increased in A549R (P=0.017, P=0.001, P=0.000) and H1299R (P=0.033, P=0.000, P=0.008) cells, respectively;the values of α and α/β were significantly reduced in A549R (P=0.018;P=0.007) and H1299R (P=0.001;P=0.009) cells, respectively. The survival rates in A549R and H1299R cells after exposure to radiation with various doses were significantly higher than those in the control groups (all P<0.05). After exposure, the apoptosis rates were significantly reduced in A549R and H1299R cells (P=0.02, P=0.01);the invasion and migration rates were significantly increased in A549R (P=0.000;P=0.001) and H1299R (P=0.001, P=0.002) cells;the expression of E-cadherin was significantly down-regulated in A549R and H1299R cells (P=0.00, P=0.01), while the expression of vimentin was significantly elevated in A549R and H1299R cells (P=0.02, P=0.01). Conclusions The radiation-resistant lung carcinoma cell lines are successfully established. Both cell lines show enhanced invasion and migration, which may be associated with EMT.
Qiu Xiangnan,Zhang Wei,Li Hao et al. Establishment and biological characterization of radiation-resistant lung carcinoma cell lines[J]. Chinese Journal of Radiation Oncology, 2015, 24(6): 703-707.
[1] 国家癌症中心,卫生部疾病预防控制局.2013中国肿瘤登记年报[M].北京:军事医学科学出版社,2014. [2] Yang HJ,Kim N,Seong KM,et al. Investigation of radiation-induced transcriptome profile of radioresistant non-small cell lung cancer A549 cells using RNA-seq[J/OL].PLoS ONE,2013,8(3):e59319[2014-10-10].http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0059319.DOI:10.1371/journal.pone.0059319. [3] Moncharmont C,Levy A,Gilormini M,et al. Targeting a cornerstone of radiation resistance:cancer stem cell[J].Cancer Letter,2012,322(2):139-147.DOI:10.1016/j.canlet.2012.03.024. [4] Grosso S,Doyen J,Parks SK,et al. MiR-210 promotes a hypoxic phenotype and increases radioresistance in human lung cancer cell lines[J/OL].Cell Death Dis,2013,4:e544.http://www.nature.com/cddis/journal/v4/n3/full/cddis201371a.html. DOI:10.1038/cddis.2013.71. [5] Lee YS,Oh JH,Yoon S,et al. Differential gene expression profiles of radioresistant non–small-cell lung cancer cell lines established by fractionated irradiation:tumor protein p53-inducible protein 3 confers sensitivity to ionizing radiation[J].Int J Radiat Oncol Biol Phys,2010,77(3):858-866.DOI:10.1016/j.ijrobp.2009.12.076. [6] 李果,刘勇,粟忠武,等.放射线照射诱导鼻咽癌细胞上皮-间质转化的作用研究[J].中华耳鼻咽喉头颈外科杂志,2013,48(8):662-667. [7] Meng MB,Wang HH,Guo WH,et al. Targeting pyruvate kinase M2 contributes to radiosensitivity of non-small cell lung cancer cells in vitro and in vivo[J].Cancer Lett,2014,356(2):985-993.DOI:10.1016/j.canlet.2014.11.016. [8] Li J,Yang CX,Mei ZJ,et al. Involvement of cdc25c in cell cycle alteration of a radioresistant lung cancer cell line established with fractionated ionizing radiation[J].Asian Pac J Cancer Prev,2013,14(10):5725-5730. [9] Lee SY,Park HR,Rhee J,et al. Therapeutic effect of oncolytic adenovirus expressing relaxin in radioresistant oral squamous cell carcinoma[J].Oncol Res,2013,20(9):419-425.DOI:10.3727/096504013X13657689383139. [10] Hardee ME,Marciscano AE,Medina-Ramirez CM,et al. Resistance of glioblastoma initiating cells to radiation mediated by the tumor microenvironment can be abolished by inhibiting transforming growth factor-β(TGFβ)[J].Cancer Res,2012,72(16):4119-4129.DOI:10.1158/0008－5472.CAN-12-0546. [11] Gomez-Casal R,Bhattacharya C,Ganesh N,et al. Non-small cell lung cancer cells survived ionizing radiation treatment display cancer stem cell and epithelial-mesenchymal transition phenotypes[J].Mol Cancer,2013,12(1):94.DOI:10.1186/1476－4598-12-94. [12] Sofia Vala I,Martins LR,Imaizumi N,et al. Low doses of ionizing radiation promote tumor growth and metastasis by enhancing angiogenesis[J/OL].PloS One,2010,5(6):e11222.http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0011222.DOI:10.1371/journal.pone.0011222. [13] Karamitopoulou E.Role of epithelial-mesenchymal transition in pancreatic ductal adenocarcinoma:is tumor budding the missing link?[J].Front Oncol,2013,3:221.DOI:10.3389/fonc.2013.00221. [14] Kalluri R,Weinberg RA.The basics of epithelial-mesenchymal transition[J].J Clin Invest,2009,119(6):1420-1428.DOI:10.1172/JCI39104. [15] Thiery JP,Lim CT.Tumor Dissemination:an EMT affair[J].Cancer Cell,2013,23(3):272-273.DOI:10.1016/j.ccr.2013.03.004. [16] Jordan NV,Johnson GL,Abell AN.Tracking the intermediate stages of epithelial-mesenchymal transition in epithelial stem cells and cancer[J].Cell Cycle,2011,10(17):2865-2873. [17] Scanlon CS,Van Tubergen EA,Inglehart RC,et al. Biomarkers of epithelial-mesenchymaltransition in squamous cell carcinoma[J].J Dent Res,2013,92(2):114-121.DOI:10.1177/0022034512467352.
2015, Vol. 24 
