VEGF-A/Neuropilin 1 Pathway Confers Cancer Stemness via Activating Wnt/β-Catenin Axis in Breast Cancer Cells

Abstract

Background/Aims: Targeting cancer stem cells (CSCs) is emerging as a promising method for cancer treatment. We previously indicated that knockdown of Neuropilin 1(NRP-1) could inhibit breast cancer cell proliferation. Here, we continue exploring the roles and mechanisms of VEGF-A/NRP-1 axis in breast CSCs formation. Methods: qRT-PCR was used to detect the levels of VEGF-A and NRP-1 in breast cancer sphere cells and wild-type cells. Mammospheres formation, flow cytometry, soft agar colony and tumor formation assays were performed to evaluate the effects of VEGF-A/NRP-1 on breast cancer stemness. Further HUVECs tube formation, cell invasion assays were carried out to detect the effects of VEGF-A/NRP-1 on breast cancer spheres-induced angiogenesis. Finally, Annexin V/PI apoptosis and CCK8 assays were used to detect the effects of VEGF-A/NRP-1 on chemoresistance. Results: Overexpression of VEGF-A or NRP-1 conferred CSCs-related traits in MCF-7 cells, while knockdown of VEGF-A or NRP-1 reduced CSCs-related traits in MDA-MB-231 cells in vitro and in vivo. Notably, VEGF-A acted in a NRP-1 dependent way. Mechanistically, the VEGF-A/NRP-1 axis conferred CSCs phenotype via activating Wnt/β-catenin pathway. Conclusion: our results suggest that VEGF-A/NRP-1 axis could confer CSCs-related traits and chemoresistance.

miR-376a inhibits breast cancer cell progression by targeting neuropilin-1 NR

“…Notably, a previous research has reported that peptides inhibiting the binding of VEGF-A/NRP-1 could inhibit breast cancer progression,9 and two groups have demonstrated that a monoclonal antibody targeting neuropilin-1 or a neuropilin-1 antagonist could exert anticancer effects in breast cancer via in vitro and in vivo experiments 10,11. Our studies have previously shown that RNA interference-mediated NRP-1 silencing could inhibit breast cancer cell proliferation and promote cell apoptosis, and VEGF-A/NRP-1 pathway could confer cancer stemness via activating Wnt/β-Catenin axis in breast cancer cells 12,13. However, the mechanisms by which NRP-1 is regulated in breast cancer cells are still not clear.…”

“…Since other and our groups had previously reported the promotive effects of NRP-1 on breast cancer progression,7,8,12,13 here we sought to reveal the mechanisms by which NRP-1 was regulated in breast cancer cells. miRNA.org (http://34.236.212.39/microrna/getMrna.do?gene=8829&utr=20945&organism=9606) was used to predict the potential miRNAs that could target NRP-1.…”

“…The NRP-1 overexpression lentivirus vector was constructed in our previous study and referred to as LV-NRP-1 12. We thank RiboBio Inc. (Guangzhou, China) for constructing miR-376a overexpression lentivirus vector, LV-miR-376a and miR-control lentivirus (control).…”

“…The detailed procedure is mentioned in our previous study 12. The primary antibodies against NRP-1 (ab81321), E-cadherin (ab1416), Vimentin (ab8978), Wnt3a (ab28472), β-catenin (ab32572), anti-cleaved caspase 3 (ab32042), anti-cleaved PARP (ab32064), Caspase 3 (ab13847), PARP (ab74290) and β-actin (ab8226) were purchased from Abcam (Cambridge, UK).…”

HOXD3 Plays a Critical Role in Breast Cancer Stemness and Drug Resistance

“…Acquired resistance is thought to occur when the subpopulation of CSCs that survives a course of treatment accumulate mutations that confer a drug resistance phenotype [21, 22]. Breast cancer stem cells have specifically been shown to acquire therapeutic resistance through a variety of signaling pathways, including those dependent on MEK/ERK [23], TGF-β [24], VEGF [25] and Wnt/β-catenin [26]. There are a variety of genes, including homeobox genes, that play crucial roles in tumorigenesis and that do so by interacting with signaling pathways [27, 28].…”