Neuropilin-1 Knockout and Rescue Confirms Its Role to Promote Metastasis in MDA-MB-231 Breast Cancer Cells

Abstract

Breast cancer (BC) metastasis remains a leading cause of female mortality. Neuropilin-1 (NRP-1) is a glycoprotein receptor that plays ligand-dependent roles in BC. Clinical studies indicate its correlation with metastatic disease; however, its functional role in BC metastasis remains uncertain. CRISPR-Cas9 was used to knockout the NRP-1 gene in MDA-MB-231 BC cells, and the effects on metastasis were determined using an orthotopic mouse engraftment model. NRP-1 expression in knockout cells was rescued using a recombinant cDNA with a silent mutation in the sgRNA target-adjacent PAM sequence. Differentially expressed genes between NRP-1 knockout and control cells were determined using whole-transcriptome sequencing and validated using real-time PCR. NRP-1KO cells showed a pronounced reduction in the metastasis to the lungs. KEGG pathway analysis of the transcriptome data revealed that PI3K and ECM receptor interactions were among the top altered pathways in the NRP-1KO cells. In addition, reduction in metastasis enhancers proteins, Integrin-β3 and Tenascin-C, and genes CCL20 and FN1 and upregulation of metastasis suppressor genes, ACVRL and GPX3 in NRP-1KO were detected. These findings provide evidence for a functional role for NRP-1 in BC metastasis, supporting further exploration of NRP-1 and the identified genes as targets in treating metastatic BC.

Identification of key modules and hub genes involved in regulating the feather follicle development of Wannan chickens using WGCNA

“…The ECM-receptor interaction signaling pathway plays an important role in tumor shedding, adhesion, degradation, movement, and proliferation ( Bao, et al, 2019 ; Huang, et al, 2024 ). The role of ECM in other cancers has also as well been discovered ( Cui, et al, 2018 ; Yan, et al, 2018 ; Zhang, et al, 2022 ; Al-Zeheimi, et al, 2023 ). However, the ECM-receptor interaction signaling pathway has also been found to play an important role in feather follicle development.…”

Low NT5DC2 expression predicts favorable prognosis and suppresses soft tissue sarcoma progression via ECM-receptor interaction pathway

“…Zhang et al [30] reported that the activation of the ECM receptor interaction pathway can promote hepatocellular carcinoma proliferation and invasion. In breast cancer, the ECM-receptor interaction pathway plays an important role in cancer progression and metastasis [31] . The important functions of the ECM-receptor interaction pathway in sarcoma have also been confirmed.…”

Natural Product Cordycepin (CD) Inhibition for NRP1/CD304 Expression and Possibly SARS-CoV-2 Susceptibility Prevention on Cancers

“…These specific domains participated in different signaling pathways and versatile roles controlling survival, migration, and invasion, as well as angiogenesis and axon guidance, through binding ligands to co-receptors, including VEGF and semaphorin family members [3,4]. For example, in breast cancer cells MDA-MB-231, CRISPR-Cas9 knocking out the NRP1 gene was reported to have a pronounced reduction in lung metastasis [6].…”