Department of Histology and Embryology, Xiangya School Medicine, Central South University, Changsha, 410008, China.
J Cancer Res Clin Oncol. 2012 Dec;138(12):2117-25. doi: 10.1007/s00432-012-1293-0. Epub 2012 Jul 31.
To identify the proteins involved in radioresistance in nasopharyngeal cancer (NPC) cells.
Sublethal ionizing radiation was applied to establish a radioresistant NPC cell line from its parental NPC cell line CNE1. Clonogenic survival assay, cell growth assay and flow cytometry analysis were used to examine the difference of radiosensitivity in the radioresistant CNE1 cells (CNE1-IR) and control CNE1 cells. Comparative proteomics was performed to identify the differential proteins in the two cell lines. Association of HSP27, one of upregulated proteins in CNE1-IR cells, with NPC cell radioresistance was selected for further investigation using antisense oligonucleotides (ASOs), clonogenic survival assay, Hoechst 33258 staining of apoptotic cells and MTT assay of cell viability.
Radioresistant NPC cell line CNE1-IR derived from its parental cell line CNE1 was established. Thirteen differential proteins in the CNE1-IR and CNE1 cells were identified by proteomics, and differential expression of HSP27, one of identified proteins, was selectively confirmed by western blot. Inhibition of HSP27 expression by HSP27 ASOs decreased clonogenic survival and cell viability and increased cell apoptosis of CNE1-IR cells after irradiation, that is, enhanced radiosensitivity of CNE1-IR cells.
The data suggest that HSP27 is a radioresistant protein in NPC cells, and its upregulation may be involved in the NPC radioresistance.
鉴定鼻咽癌（NPC）细胞辐射抗性相关的蛋白。
亚致死剂量电离辐射用于建立其亲本 NPC 细胞系 CNE1 的耐辐射 NPC 细胞系。集落形成实验、细胞生长实验和流式细胞术分析用于检测耐辐射 CNE1 细胞（CNE1-IR）和对照 CNE1 细胞之间放射敏感性的差异。采用比较蛋白质组学方法鉴定两种细胞系中的差异蛋白。选择 HSP27 作为 CNE1-IR 细胞中上调的蛋白之一，用反义寡核苷酸（ASOs）、凋亡细胞 Hoechst 33258 染色和细胞活力 MTT 检测进一步研究其与 NPC 细胞放射抗性的关系。
建立了源自亲本细胞系 CNE1 的耐辐射 NPC 细胞系 CNE1-IR。通过蛋白质组学鉴定出 CNE1-IR 和 CNE1 细胞中的 13 种差异蛋白，并通过 Western blot 选择性地确认了 HSP27 这一鉴定出的蛋白之一的差异表达。HSP27 ASOs 抑制 HSP27 表达可降低 CNE1-IR 细胞的集落形成能力和细胞活力，增加照射后细胞凋亡，即增强 CNE1-IR 细胞的放射敏感性。
数据表明 HSP27 是 NPC 细胞中的一种辐射抗性蛋白，其上调可能参与 NPC 的放射抗性。
