Chen Zhen, Long Lu, Wang Kun, Cui Facai, Zhu Lepan, Tao Ya, Wu Qiong, Xiang Manlin, Liang Yunlai, Qiu Shiyang, Xiao Zhiqiang, Yi Bin
Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
The Higher Educational Key Laboratory for Cancer Proteomics and Translational Medicine of Hunan Province, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Oncotarget. 2016 Jun 7;7(23):34022-37. doi: 10.18632/oncotarget.9067.
To identify metastasis-related proteins in nasopharyngeal carcinoma (NPC), iTRAQ-tagging combined with 2D LC-MS/MS analysis was performed to identify the differentially expressed proteins (DEPs) in high metastatic NPC 5-8F cells and non-metastatic NPC 6-10B cells, and qRT-PCR and Western blotting were used to confirm DEPs. As a result, 101 DEPs were identified by proteomics, and 12 DEPs were selectively validated. We further detected expression of three DEPs (RAN, SQSTM1 and TRIM29) in a cohort of NPC tissue specimens to assess their value as NPC metastatic biomarkers, and found that combination of RAN, SQSTM1 and TRIM29 could discriminate metastatic NPC from non-metastatic NPC with a sensitivity of 88% and a specificity of 91%. TRIM29 and RAN expression level were closely correlated with lymph node and distant metastasis and clinical stage (P <0.05) in NPC patients. Finally, a combination of loss-of-function and gain-of-function approaches was performed to determine the effects of TRIM29 on NPC cell proliferation, migration, invasion and metastasis. The results showed that TRIM29 knockdown significantly attenuated while TRIM29 overexpression promoted NPC cell in vitro proliferation, migration and invasion and in vivo metastasis. The present data first time show that SQSTM1, RAN and TRIM29 are novel potential biomarkers for predicting NPC metastasis, demonstrate that TRIM29 is a metastasis-promoted protein of NPC.
为了鉴定鼻咽癌（NPC）中与转移相关的蛋白质，采用iTRAQ标记结合二维液相色谱-串联质谱（2D LC-MS/MS）分析来鉴定高转移NPC 5-8F细胞和低转移NPC 6-10B细胞中差异表达的蛋白质（DEPs），并通过qRT-PCR和蛋白质免疫印迹法对DEPs进行验证。结果，通过蛋白质组学鉴定出101个DEPs，并选择性验证了12个DEPs。我们进一步检测了一组NPC组织标本中三种DEPs（RAN、SQSTM1和TRIM29）的表达，以评估它们作为NPC转移生物标志物的价值，发现RAN、SQSTM1和TRIM29的联合检测能够区分转移性NPC和非转移性NPC，灵敏度为88%，特异性为91%。在NPC患者中，TRIM29和RAN的表达水平与淋巴结转移、远处转移及临床分期密切相关（P<0.05）。最后，采用功能丧失和功能获得相结合的方法来确定TRIM29对NPC细胞增殖、迁移、侵袭和转移的影响。结果表明，敲低TRIM29可显著减弱NPC细胞的体外增殖、迁移和侵袭以及体内转移，而过表达TRIM29则可促进这些过程。本研究首次表明，SQSTM1、RAN和TRIM29是预测NPC转移的新型潜在生物标志物，证明TRIM29是一种促进NPC转移的蛋白质。
