Annexin A1 May Induce Pancreatic Cancer Progression as a Key Player of Extracellular Vesicles Effects as Evidenced in the In Vitro MIA PaCa-2 Model System

Abstract

Pancreatic Cancer (PC) is one of the most aggressive malignancies worldwide. As annexin A1 (ANXA1) is implicated in the establishment of tumour metastasis, the role of the protein in PC progression as a component of extracellular vesicles (EVs) has been investigated. EVs were isolated from wild type (WT) and ANXA1 knock-out (KO) PC cells and then characterised by multiple approaches including Western blotting, Field Emission-Scanning Electron Microscopy, and Dynamic Light Scattering. The effects of ANXA1 on tumour aggressiveness were investigated by Wound-Healing and invasion assays and microscopic analysis of the Epithelial to Mesenchymal Transition (EMT). The role of ANXA1 on angiogenesis was also examined in endothelial cells, using similar approaches. We found that WT cells released more EVs enriched in exosomes than those from cells lacking ANXA1. Notably, ANXA1 KO cells recovered their metastatic potential only when treated by WT EVs as they underwent EMT and a significant increase of motility. Similarly, human umbilical vein endothelial cells (HUVEC) migrated and invaded more rapidly when treated by WT EVs whereas ANXA1 KO EVs weakly induced angiogenesis. This study suggests that EVs-related ANXA1 is able to promote cell migration, invasion, and angiogenesis, confirming the relevance of this protein in PC progression.

Extracellular Vesicles (EVs) and Pancreatic Cancer: From the Role of EVs to the Interference with EV-Mediated Reciprocal Communication

“…A recent finding indicated that ANXA1 can be incorporated into EVs. The ANXA1-containing EVs leads to an activation of the EMT process, along with the increased motility of exosome-receiving PCCs [43] (Figure 1 and Table 1). However, another study indicated the opposing role of ANXA1 in PCCs.…”

“…As stated in Section 2.2.2, ANXA1 can be incorporated in PCC-derived EVs and modulate the motility of cancer cells. Beyond this, it was unequivocally addressed that ANXA1 in PCC-derived EVs can activate ECs, and it was ascertained by monitoring proliferation, migration, invasion and tube formation of endothelial cells [43] (Figure 2 and Table 2).…”

“…It was investigated that the knockdown of ANXA1 diminishes the amounts of secreted EVs, particularly exosomes, in pancreatic cancer cells, indicating that ANXA1 positively regulates exosome biosynthesis [43]. ANXA1 is known to create the membrane contact sites and inward vesiculation, which are required for the formation of intraluminal vesicles in multivesicular bodies (MVBs) [136,137].…”

Involvement of Extracellular Vesicles in Vascular-Related Functions in Cancer Progression and Metastasis

“…The involvement of cancer-derived exosomes in EndMT remains unclear; however, relevant data are gradually accumulating, shedding light on this mechanism. For instance, it has been reported that exosomal Annexin A1 and RAC1/PAK2 induce EndEMT, whereas miRNA-200 suppresses EndEMT [85,112,113].…”

PCL/Mesoglycan Devices Obtained by Supercritical Foaming and Impregnation

“…A 24-well plate was coated with Matrigel (Becton Dickinson Labware, Franklin Lakes, NJ, USA) mixed to EGM-2 1:1 on ice and incubated at 37 • C for 30 min to allow gelation to occur, as reported in Pessolano et al, 2018 [28]. HUVEC cells were added to the top of the gel at a density of 2 × 10 4 cells/well in the presence or not of sodium MSG and PCL-derived MSG.…”