Surgical Department of Head and Neck, The Third Affiliated Hospital, Kunming Medical University, Kunming, Yunnan, P. R. China.
Proteomics. 2011 Jul;11(14):2911-20. doi: 10.1002/pmic.201000483.
Metastasis to secondary sites remains the leading cause of nasopharyngeal carcinoma (NPC)-associated death. In order to identify the candidate protein(s) responsible for the differential metastatic capacity, the protein expression profiling between NPC cell line CNE-2 and its highly metastatic subclone S-18 were compared by 2-DE. In total, 18 spots were differentially expressed between these two cell lines. Among all, seven proteins were identified with further MS analysis. Western blotting further validated upregulation of HSP27 and ezrin, and downregulation of valosin containing protein and keratin 18 in S-18. Moreover, the knockdown of HSP27 was found to significantly decrease the invasive ability of S-18. On the other hand, overexpression of HSP27 in NP460 cells, which generated little endogenous HSP27 and less invasive, was noted to gain enhanced metastatic capability. Real-time PCR confirmed that the transcriptional levels of NF-κB and MMP9, MMP11 were downregulated after inhibition of HSP27 in S-18, which implicated that HSP27 enhanced the metastatic property of NPC cells probably via the NF-κB-mediated activation of MMPs. The findings in this work provided us a platform for further elucidating the underlying mechanisms of NPC metastasis and demonstrated that HSP27 would be a valid target for anti-cancer drug development.
转移到继发部位仍然是导致鼻咽癌（NPC）相关死亡的主要原因。为了确定导致差异转移能力的候选蛋白，通过 2-DE 比较 NPC 细胞系 CNE-2 和其高转移性亚克隆 S-18 之间的蛋白质表达谱。在这两种细胞系之间，总共鉴定出 18 个差异表达的斑点。其中，通过 MS 分析进一步鉴定了 7 种蛋白质。Western blotting 进一步验证了 HSP27 和 ezrin 的上调，以及 valosin 蛋白和角蛋白 18 的下调在 S-18 中。此外，发现 HSP27 的敲低显著降低了 S-18 的侵袭能力。另一方面，在 NP460 细胞中过表达 HSP27，该细胞产生的内源性 HSP27 较少，侵袭性较弱，被认为获得了增强的转移能力。实时 PCR 证实，抑制 S-18 中的 HSP27 后，NF-κB 和 MMP9、MMP11 的转录水平下调，这表明 HSP27 可能通过 NF-κB 介导的 MMPs 激活增强 NPC 细胞的转移特性。本研究的结果为我们进一步阐明 NPC 转移的潜在机制提供了一个平台，并表明 HSP27 将成为抗癌药物开发的有效靶点。
