Im Chang-Nim, Kim Byeong Mo, Moon Eun-Yi, Hong Da-Won, Park Joung Whan, Hong Sung Hee
Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, Seoul 139-706, Korea.
Department of Bioscience and Biotechnology, Sejong University, Seoul 143-747, Korea.
Genomics Inform. 2013 Dec;11(4):245-53. doi: 10.5808/GI.2013.11.4.245. Epub 2013 Dec 31.
A radioresistant cell line was established by fractionated ionizing radiation (IR) and assessed by a clonogenic assay, flow cytometry, and Western blot analysis, as well as zymography and a wound healing assay. Microarray was performed to profile global expression and to search for differentially expressed genes (DEGs) in response to IR. H460R cells demonstrated increased cell scattering and acidic vesicular organelles compared with parental cells. Concomitantly, H460R cells showed characteristics of increased migration and matrix metalloproteinase activity. In addition, H460R cells were resistant to IR, exhibiting reduced expression levels of ionizing responsive proteins (p-p53 and γ-H2AX); apoptosis-related molecules, such as cleaved poly(ADP ribose) polymerase; and endoplasmic reticulum stress-related molecules, such as glucose-regulated protein (GRP78) and C/EBP-homologous protein compared with parental cells, whereas the expression of anti-apoptotic X-linked inhibitor of apoptosis protein was increased. Among DEGs, syntrophin beta 2 (SNTB2) significantly increased in H460R cells in response to IR. Knockdown of SNTB2 by siRNA was more sensitive than the control after IR exposure in H460, H460R, and H1299 cells. Our study suggests that H460R cells have differential properties, including cell morphology, potential for metastasis, and resistance to IR, compared with parental cells. In addition, SNTB2 may play an important role in radioresistance. H460R cells could be helpful in in vitro systems for elucidating the molecular mechanisms of and discovering drugs to overcome radioresistance in lung cancer therapy.
通过分次电离辐射（IR）建立了一种耐辐射细胞系，并通过克隆形成试验、流式细胞术、蛋白质免疫印迹分析、酶谱分析和伤口愈合试验进行评估。进行基因芯片分析以描绘全局表达情况，并寻找对IR有差异表达的基因（DEG）。与亲代细胞相比，H460R细胞表现出细胞散射增加和酸性囊泡细胞器增多。同时，H460R细胞表现出迁移增加和基质金属蛋白酶活性增强的特征。此外，H460R细胞对IR具有抗性，与亲代细胞相比，其电离反应蛋白（p-p53和γ-H2AX）、凋亡相关分子（如裂解的聚（ADP核糖）聚合酶）以及内质网应激相关分子（如葡萄糖调节蛋白（GRP78）和C/EBP同源蛋白）的表达水平降低，而抗凋亡的X连锁凋亡抑制蛋白的表达增加。在DEG中，肌养蛋白β2（SNTB2）在H460R细胞中对IR的反应显著增加。在H460、H460R和H1299细胞中，IR暴露后，用小干扰RNA（siRNA）敲低SNTB2比对照组更敏感。我们的研究表明，与亲代细胞相比，H460R细胞具有不同的特性，包括细胞形态、转移潜能和对IR的抗性。此外，SNTB2可能在辐射抗性中起重要作用。H460R细胞可能有助于体外系统阐明肺癌治疗中辐射抗性的分子机制并发现克服辐射抗性的药物。
