Higher levels of TIMP-1 expression are associated with a poor prognosis in triple-negative breast cancer.
Higher levels of TIMP-1 expression are associated with a poor prognosis in triple-negative breast cancer.
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较高水平的 TIMP-1 表达与三阴性乳腺癌的不良预后相关
DOI：
10.1186/s12943-016-0515-5
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发表时间：
2016-04-30
期刊：
影响因子：
37.3
通讯作者：
Sun X
中科院分区：
文献类型：
--
作者：
Cheng G;Fan X;Hao M;Wang J;Zhou X;Sun X
研究背景金属蛋白酶组织抑制剂-1（TIMP-1）是一种多功能蛋白质，可直接调控细胞凋亡和肿瘤转移。在这项研究中，我们研究了TIMP-1影响三阴性乳腺癌（TNBC）的功能和分子机制。采用Kaplan-Meier分析评估了根据TIMP-1表达水平分层的不同分子亚型乳腺癌患者的总生存期。硫酸氢盐测序PCR（BSP）分析TIMP-1启动子甲基化状态。实时-PCR（RT-PCR），蛋白质印迹和ELISA测定用于评估基因和蛋白质在细胞系和人体组织标本中的表达。此外，TIMP-1的功能进行了分析，使用一系列的在体外和体内试验与细胞中，TIMP-1被抑制使用RNAi或中和antibodies.ResultsWe发现，血清TIMP-1水平强烈增强与TNBC患者和TIMP-1水平升高与TNBC预后不良。然而，TIMP-1水平与其他亚型乳腺癌或乳腺癌患者总人群的总生存期无显著相关性。我们还报告了第一个证据表明，与非TNBC细胞系相比，TIMP-1启动子在TNBC细胞系中是低甲基化的，这表明TNBC中TIMP-1的异常表达是由DNA甲基化降低引起的。RNAi介导的TNBC细胞中TIMP-1的沉默诱导细胞周期停滞在G1期并降低细胞周期蛋白D1的表达。此外，机制分析显示，p-Akt和p-NF-κB信号通路，而不是GSK-3β和MAPK 1/2通路，与TNBC细胞中TIMP-1过表达相关。此外，针对TIMP-1的中和抗体显着降低了肿瘤生长的速度在vivo.ConclusionsOur的研究结果表明，TIMP-1是一个生物标志物，表明在TNBC患者的预后不良，靶向TIMP-1可以提供一个有吸引力的治疗干预，特别是三阴性乳腺癌患者。
BackgroundTissue inhibitor of metalloproteinases-1 (TIMP-1) is a multifunctional protein that can directly regulate apoptosis and metastasis. In this study, we investigated the functional and molecular mechanisms by which TIMP-1 influences triple-negative breast cancer (TNBC).MethodsThe expression level of TIMP-1 in breast cancer tissues was analyzed using the ONCOMINE microarray database. The overall survival of patients with distinct molecular subtypes of breast cancer stratified by TIMP-1 expression levels was evaluated using Kaplan–Meier analysis. Bisulfate sequencing PCR (BSP) was used to analyze the methylation status of the TIMP-1 promoter. Real-time-PCR (RT-PCR), Western blot and ELISA assays were used to evaluate gene and protein expression in cell lines and human tissue specimens. In addition, TIMP-1 function was analyzed using a series of in vitro and in vivo assays with cells in which TIMP-1 was inhibited using RNAi or neutralizing antibodies.ResultsWe found that serum TIMP-1 levels were strongly enhanced in patients with TNBC and that elevated TIMP-1 levels were associated with a poor prognosis in TNBC. However, TIMP-1 levels were not significantly associated with overall survival in other subtypes of breast cancer or in the overall population of breast cancer patients. We also report the first evidence that the TIMP-1 promoter is hypomethylated in TNBC cell lines compared with non-TNBC cell lines, suggesting that aberrant TIMP-1 expression in TNBC results from reduced DNA methylation. RNAi-mediated silencing of TIMP-1 in TNBC cells induced cell cycle arrest at the G1 phase and reduced cyclin D1 expression. In addition, mechanistic analyses revealed that the p-Akt and p-NF-κB signaling pathways, but not the GSK-3β and MAPK1/2 pathways, are associated with TIMP-1 overexpression in TNBC cells. Moreover, neutralizing antibodies against TIMP-1 significantly decreased the rate of tumor growth in vivo.ConclusionsOur findings suggest that TIMP-1 is a biomarker indicative of a poor prognosis in TNBC patients and that targeting TIMP-1 may provide an attractive therapeutic intervention specifically for triple-negative breast cancer patients.