DU Guifang, Hao Chengcheng, Gu Yanan, Wang Zhiyan, Jiang Wen G, He Junqi, Cheng Shan
Department of Biochemistry and Molecular Biology, Capital Medical University, Beijing, P.R. China Beijing Key Laboratory of Cancer & Metastasis Research, Capital Medical University, Beijing, P.R. China.
Department of Biochemistry and Molecular Biology, Capital Medical University, Beijing, P.R. China Beijing Key Laboratory of Cancer & Metastasis Research, Capital Medical University, Beijing, P.R. China Cardiff China Medical Research Collaborative, Cardiff University School of Medicine, Heath Park, Cardiff, U.K.
Anticancer Res. 2016 Mar;36(3):1165-73.
Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) has been reported to interact with many cancer-related proteins. We recently identified a novel NHERF1 mutation (E43G) in breast tumours.
The candidates of NHERF1 mutation were identified in breast cancer tissues by polymerase chain reaction and DNA sequencing. Wild-type NHERF1 and E43G mutation were expressed in NHERF1-knockdown cells (MCF7ΔNHERF1) and low-NHERF1-expressing cells (SKMES-1). The effects of mutated NHERF1 on cell functions were examined using in vitro methods. Glutathione S-transferase pull-down assays and western blotting were performed to study the effects of NHERF1 mutation on its interaction with cancer-related proteins.
Compared to wild-type NHERF1, expression of the mutated NHERF1 failed to suppress malignant traits in cancer cells, attenuated interaction of NHERF1 protein with epidermal growth factor receptor (EGFR), and inactivated its inhibition of EGF-induced Akt and extracellular regulated protein kinases (ERK) activation.
The results show the causal role of NHERF1 in the regulation of the EGFR pathway and the progression of breast cancer.
据报道，钠氢交换调节因子1（NHERF1）与许多癌症相关蛋白相互作用。我们最近在乳腺肿瘤中发现了一种新的NHERF1突变（E43G）。
通过聚合酶链反应和DNA测序在乳腺癌组织中鉴定NHERF1突变的候选者。野生型NHERF1和E43G突变在NHERF1敲低细胞（MCF7ΔNHERF1）和低NHERF1表达细胞（SKMES-1）中表达。使用体外方法检测突变的NHERF1对细胞功能的影响。进行谷胱甘肽S-转移酶下拉试验和蛋白质印迹分析以研究NHERF1突变对其与癌症相关蛋白相互作用的影响。
与野生型NHERF1相比，突变的NHERF1的表达未能抑制癌细胞中的恶性特征，减弱了NHERF1蛋白与表皮生长因子受体（EGFR）的相互作用，并使其对EGF诱导的Akt和细胞外调节蛋白激酶（ERK）激活的抑制失活。
结果表明NHERF1在EGFR途径调节和乳腺癌进展中起因果作用。
