蛋白质组学方法识别nm23-H1为鼻咽癌的转移抑制蛋白和预后因子(英文)

Identification of nm23-H1 as a metastatic suppressor and prognostic factor in nasopharyngeal carcinoma by proteomic analysis

目的:筛选鼻咽癌(NPC)转移相关蛋白质,为鼻咽癌防治提供科学依据。方法:采用二维电泳和质谱技术筛选不同转移潜能鼻咽癌细胞系(5-8F和6-10B)的差异表达蛋白质,并应用Western印迹对部分差异蛋白质进行验证;使用siRNA抑制差异蛋白质nm23-H1的表达,分析nm23-H1表达水平对鼻咽癌细胞体外侵袭能力的影响;应用免疫组织化学染色分析nm23-H1表达水平与鼻咽癌临床病理特征和预后的关系。结果:在不同转移潜能鼻咽癌细胞系中鉴定了15个差异表达的蛋白质,并选择性证实3个差异蛋白质;siRNA下调nm23-H1的表达能增强6-10B鼻咽癌细胞的体外侵袭能力;淋巴结转移鼻咽癌组织中nm23-H1的水平显著低于原发性鼻咽癌;nm23-H1的表达水平与鼻咽癌淋巴结和远处转移、临床分期和复发正相关;生存分析显示nm23-H1低表达的鼻咽癌患者预后差;多因素分析显示nm23-H1表达水平是鼻咽癌患者独立的预后因子。结论:nm23-H1是鼻咽癌的转移抑制蛋白和预后因子。

Objective： To identify proteins associated with nasopharyngeal carcinoma（NPC） metastasis,and provide scientific basis for the prevention and cure of NPC. Methods： A two-dimensional gel electrophoresis and mass spectrometry were performed to screen for differential proteins between highly metastatic 5-8F and non-metastatic 6-10B NPC cell lines.Western blot was used to confirm the differential proteins.We used siRNA to inhibit the expression of differential protein nm23-H1 to determine the association of nm23-H1 with NPC in vitro invasive ability.Immunohistochemistry and statistics were used to evaluate the correlation of nm23-H1 expression with clinicopathological features and clinical outcomes in paraffin-embedded archival tissues including 93 cases of primary NPC and 20 cases of cervical lymphonode metastatic NPC（LMNPC）. Results： A total of 15 differential proteins in the 2 cell lines were identified by a proteomic approach,and 3 differential proteins were selectively confirmed.Downregulation of nm23-H1 by siRNA significantly increased the in vitro invasive ability of 6-10B.Significant nm23-H1 downregulation was observed in LMNPC compared with primary NPC.nm23-H1 downregulation in primary NPC was positively correlated with lymphonode and distant metastasis,advanced clinical stage and recurrence.Survival curves showed that patients with nm23-H1 downregulation in primary NPC had a poor prognosis.Multivariate analysis confirmed that nm23-H1 expression level in primary NPC was an independent prognostic indicator.Conclusion： nm23-H1 behaves as a metastasis suppressor in NPC,and nm23-H1 downregulation is a biomarker for poor NPC prognosis.