Identification of the proteins related to p53-mediated radioresponse in nasopharyngeal carcinoma by proteomic analysis.
Identification of the proteins related to p53-mediated radioresponse in nasopharyngeal carcinoma by proteomic analysis.
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DOI：
10.1016/j.jprot.2011.02.012
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发表时间：
2011-11
影响因子：
3.3
通讯作者：
Guqing Zeng;Hong Yi;Xin-Hui Li;Hui-Ying Shi;Cui Li;Maoyu Li;Pengfei Zhang;Xue-Ping Feng;Xun-Xun Wan-Xun;Jia-Quan Qu;Yan Xu;Yi Sun;Zhu-chu Chen;Zhi-Qiang Xiao
中科院分区：
文献类型：
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作者：
Guqing Zeng;Hong Yi;Xin-Hui Li;Hui-Ying Shi;Cui Li;Maoyu Li;Pengfei Zhang;Xue-Ping Feng;Xun-Xun Wan-Xun;Jia-Quan Qu;Yan Xu;Yi Sun;Zhu-chu Chen;Zhi-Qiang Xiao
放射治疗是鼻咽癌的主要治疗手段，p53与鼻咽癌的放射敏感性密切相关，但p53介导鼻咽癌放射反应的分子机制尚不清楚。我们建立了p53基因敲低的鼻咽癌细胞系CNE 2sip 53和配对的对照细胞系CNE 2/pSUPER，为研究p53介导的鼻咽癌放射反应机制提供了细胞模型。在这项研究中，我们首先比较了CNE 2sip 53和CNE 2/pSUPER的放射敏感性的克隆形成生存试验，细胞生长试验，Hoechst 33258染色和流式细胞术分析的凋亡细胞。结果显示，CNE 2 sip 53的放射敏感性明显低于CNE 2/pSUPER，提示p53在鼻咽癌放射敏感性中起介导作用。为寻找p53介导的鼻咽癌放射反应相关蛋白，采用蛋白质组学方法分别对CNE 2sip 53和CNE 2 p/SUPER中的放射反应蛋白进行鉴定，并比较CNE 2sip 53和CNE 2 p/SUPER放射反应蛋白的差异。结果在两株细胞中共鉴定出14个差异辐射反应蛋白，其中4个蛋白经Western blot证实。其中，9个和5个蛋白质分别从CNE 2 p/SUPER和CNE 2sip 53中单独鉴定。蛋白质相互作用分析表明，仅在CNE 2 p/SUPER中发现的7个差异辐射反应蛋白与p53蛋白有关。结果提示，CNE 2 p/SUPER特异性差异蛋白可能参与p53介导的鼻咽癌放射反应，这将有助于阐明p53介导的鼻咽癌细胞放射反应机制。
Radiotherapy is the primary treatment for nasopharyngeal cancer (NPC), and p53 is closely associated with the radiosensitivity of cancer, but the molecular mechanisms of p53-mediated radioresponse in NPC remains unclear. We previously established NPC CNE2sip53 cell line with p53 knockdown and paired control cell line CNE2/pSUPER, which provides a cell model system to investigate mechanisms of p53-mediated radioresponse in NPC. In this study, we first compared the radiosensitivity of CNE2sip53 and CNE2/pSUPER by a clonogenic survival assay, cell growth assay, and Hoechst 33258 staining and flow cytometry analysis of apoptotic cells. The results showed that the radiosensitivity of CNE2sip53 was significantly lower than that of CNE2/pSUPER, indicating that p53 plays a role in mediating NPC radiosensitivity. To search for the proteins associated with the p53-mediated radioresponse in NPC, a proteomic approach was performed to identify the radioresponsive proteins in CNE2sip53 and CNE2p/SUPER, respectively, and then the difference of radioresponsive proteins in CNE2sip53 and CNE2p/SUPER was compared. As a result, 14 differential radioresponsive proteins were identified in the two cell lines, 4 proteins of which were conformed by Western blot. Among them, 9 and 5 proteins were identified solely from CNE2p/SUPER and CNE2sip53, respectively. Furthermore, protein–protein interaction analysis showed that 7 differential radioresponsive proteins identified only in CNE2p/SUPER were related to p53 protein. Our results suggest that the differential radioresponsive proteins unique to CNE2p/SUPER may be involved in p53-mediated radioresponse in NPC, which will be helpful for elucidating the mechanisms of p53-mediated NPC cellular response to radiotherapy.