Supplementary Data from ANXA1 Binds and Stabilizes EphA2 to Promote Nasopharyngeal Carcinoma Growth and Metastasis

Feng, Juan; Lu, Shan-Shan; Xiao, Ta; Huang, Wei; Yi, Hong; Zhu, Wei; Fan, Songqing; Feng, Xue-Ping; Li, Jiao-Yang; Yu, Zheng-Zheng; Gao, Song; Nie, Guo-Hui; Tang, Yao-Yun; Xiao, Zhi-Qiang

doi:10.1158/0008-5472.22425835.v1

Supplementary Data from ANXA1 Binds and Stabilizes EphA2 to Promote Nasopharyngeal Carcinoma Growth and Metastasis

https://doi.org/10.1158/0008-5472.22425835

journal contribution

posted on 2023-03-31, 03:42 authored by Juan Feng, Shan-Shan Lu, Ta Xiao, Wei Huang, Hong Yi, Wei Zhu, Songqing Fan, Xue-Ping Feng, Jiao-Yang Li, Zheng-Zheng Yu, Song Gao, Guo-Hui Nie, Yao-Yun Tang, Zhi-Qiang Xiao

<p>1.Supplementary Materials and Methods; Supplementary Table S1-4; Supplementary Figure S1-8. Table S1. The clinicopathological characteristics of 184 patients with NPC. Table S2. The primers used for the amplification of the three genes by qRT-PCR. Table S3. Correlations between the two protein expression and clinicopathological characteristics in NPC. Table S4. Univariate and cox multivariate analyses of prognostic factors for overall and disease-free survival. Figure S1. Identification of ANXA1 and Cbl as interactors of EphA2 by co-IP-mass spectrometry (MS) analysis. Figure S2. ANXA1 affects EphA2 protein but not mRNA levels in the NPC cells. Figure S3. The effect of ANXA1 deletion mutant (D28-30) on EphA2 stability. Figure S4. Models for EphA2-Cbl and EphA2-ANXA1 complexes. Figure S5. Immunoblotting showing the levels of EphA2 and ANXA1 in the endogenous ANXA1 and EphA2 knockdown (shEphA2-ANXA1) 5-8F NPC cells transfected with the indicated plasmids. Figure S6. Binding of ANXA1 to EphA2 promotes EphA2 oncogenic signaling. Figure S7. A11 suppressed the tube formation of HUVECs. Figure S8. The effect of A11 peptide on the activity of pS897-EphA2/AKT signaling pathway in the NPC cells.</p>

Funding

National Key Basic Research Program (973 Program) of China

National Natural Science Foundation of China

Shenzhen Science and Technology Program of China

History

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DOI - Is supplement to ANXA1 Binds and Stabilizes EphA2 to Promote Nasopharyngeal Carcinoma Growth and Metastasis

ARTICLE ABSTRACT

Overexpression of ANXA1 and EphA2 has been linked to various cancers and both proteins have attracted considerable attention for the development of new anticancer drugs. Here we report that ANXA1 competes with Cbl for binding EphA2 and increases its stability by inhibiting Cbl-mediated EphA2 ubiquitination and degradation in nasopharyngeal carcinoma (NPC). Binding of ANXA1 to EphA2 promoted NPC cell growth and metastasis in vitro and in vivo by elevating EphA2 levels and increasing activity of EphA2 oncogenic signaling (pS897-EphA2). Expression of ANXA1 and EphA2 was positively correlated and both were significantly higher in NPC tissues than in the normal nasopharyngeal epithelial tissues. Patients with high expression of both proteins presented poorer disease-free survival and overall survival relative to patients with high expression of one protein alone. Furthermore, amino acid residues 20-30aa and 28-30aa of the ANXA1 N-terminus bound EphA2. An 11 amino acid–long ANXA1-derived peptide (EYVQTVKSSKG) was developed on the basis of this N-terminal region, which disrupted the connection of ANXA1 with EphA2, successfully downregulating EphA2 expression and dramatically suppressing NPC cell oncogenicity in vitro and in mice. These findings suggest that ANXA1 promotes NPC growth and metastasis via binding and stabilization of EphA2 and present a strategy for targeting EphA2 degradation and treating NPC with a peptide. This therapeutic strategy may also be extended to other cancers with high expression of both proteins. These findings show that EphA2 is a potential target for NPC therapeutics and an ANXA1-derived peptide suppresses NPC growth and metastasis.