Intracellular targeting of annexin A2 inhibits tumor cell adhesion, migration, and in vivo grafting

Abstract

Cytoskeletal-associated proteins play an active role in coordinating the adhesion and migration machinery in cancer progression. To identify functional protein networks and potential inhibitors, we screened an internalizing phage (iPhage) display library in tumor cells, and selected LGRFYAASG as a cytosol-targeting peptide. By affinity purification and mass spectrometry, intracellular annexin A2 was identified as the corresponding binding protein. Consistently, annexin A2 and a cell-internalizing, penetratin-fused version of the selected peptide (LGRFYAASG-pen) co-localized and specifically accumulated in the cytoplasm at the cell edges and cell-cell contacts. Functionally, tumor cells incubated with LGRFYAASG-pen showed disruption of filamentous actin, focal adhesions and caveolae-mediated membrane trafficking, resulting in impaired cell adhesion and migration in vitro. These effects were paralleled by a decrease in the phosphorylation of both focal adhesion kinase (Fak) and protein kinase B (Akt). Likewise, tumor cells pretreated with LGRFYAASG-pen exhibited an impaired capacity to colonize the lungs in vivo in several mouse models. Together, our findings demonstrate an unrecognized functional link between intracellular annexin A2 and tumor cell adhesion, migration and in vivo grafting. Moreover, this work uncovers a new peptide motif that binds to and inhibits intracellular annexin A2 as a candidate therapeutic lead for potential translation into clinical applications.

Structural and functional characterization of β2‐glycoprotein I domain 1 in anti‐melanoma cell migration

“…Clinical studies have shown that anxA2 is highly expressed in different tumor types . Accordingly, recent reports have shown the functional role for anxA2 in regulation of cell adhesion, migration, and invasion . In the present study, binding of β 2 ‐GPI to anxA2 was confirmed by immunofluorescence and coimmunoprecipitation assays.…”

“…36,37 Accordingly, recent reports have shown the functional role for anxA2 in regulation of cell adhesion, migration, and invasion. 38,39 In the present study, binding of β 2 -GPI to anxA2 was confirmed by immunofluorescence and coimmunoprecipitation assays. Furthermore, interaction between specific amino acids of β 2 -GPI D1 and anxA2 was shown by the molecular docking approach.…”

Annexin a2 as a target protein for chlorogenic acid in human lung cancer A549 cells

“…Moreover, the specific relationship between the above processes of ANXA2 has not been clearly clarified. The involvement of ANXA2 in these processes may be related to its own interaction with other skeletal proteins (39). In combination with our work, it is also likely to significantly inhibit the migration and cell cycle of tumor cells by blocking the original binding of ANXA2 and actin and other cytoskeleton proteins.…”

Mechanistic insights into Nav1.7‐dependent regulation of rat prostate cancer cell invasiveness revealed by toxin probes and proteomic analysis

“…Of the 64 differentially expressed proteins, eight proteins, Annexin A1, Annexin A2, Confilin‐1, Fascin, Muskelin, Moesin, Calcyclin‐binding protein, and High mobility group protein B1 (Table S3), are cytoskeleton proteins or related regulators, and they are actin‐associated proteins as revealed by KEGG analysis. It has accepted that the actin cytoskeleton is crucial to cell adhesion and migration . Studies have indicated that these proteins play important roles in cancer metastasis.…”