Dataset Information

Semaphorin 5A modulates focal adhesion pathway and lamellipodia formation in melanoma

ABSTRACT: Introduction Metastatic potential and rising incidence make cutaneous melanoma a major health concern worldwide. Although the therapies currently in use have proven effective, their use is limited by side effects arising and development of drug resistance. For this reason, research for new therapeutic targets and predictive biomarkers is crucial. Semaphorin 5A (SEMA5A) is a member of the Semaphorin family with involvement in cancer biology and neural development. We previously demonstrated that this protein promotes different in vitro melanoma aggressive features. In this work we further investigated the role and the mechanism of action of SEMA5A in melanoma formation and progression. Methods We performed proteomic analysis of stable knockdown SEMA5A human melanoma cells, and in silico analysis to identify cellular pathways modulated by SEMA5A. In vitro clonogenic assay and cell viability assay under different substrate conditions were conducted by using both human and murine melanoma SEMA5A knockdown clones. Western blotting analysis and immunofluorescence have been employed to investigate focal adhesion pathway regulation and lamellipodia formation after SEMA5A depletion. Nocodazole assay was used to evaluate focal adhesion dynamic. Xenograft mouse model of melanoma was used to investigate the effect of SEMA5A depletion in tumor formation and growth. Results Ingenuity pathway analysis showed that SEMA5A effects several different cellular pathways essential for cell viability, cell proliferation, and cytoskeletal dynamics, including focal adhesion and lamellipodia formation. We demonstrated that SEMA5A depletion reduced clonogenic properties and cell viability of cells in both low and high stiffness condition of culture. SEMA5A affected Focal adhesion kinase and Paxillin phosphorylation, integrin beta 1 activation, thus regulating focal adhesion dynamics, and lamellipodia formation, regulating Arp3 nucleation and expression. Delay of tumor formation and reduced tumor growth was observed in mice bearing tumor expressing low level of SEMA5A. Conclusions These findings demonstrate that SEMA5A is a novel player of melanoma aggressiveness and progression, being involved in modulating cell viability, focal adhesion signaling and lamellipodia in vitro and tumor growth in vivo.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Skin Of Leg

DISEASE(S): Melanoma

SUBMITTER: Maria Concetta Cufaro

LAB HEAD: Piero Del Boccio

PROVIDER: PXD065661 | Pride | 2025-12-01

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20221111_A375_clone5_CTRL_01_50ug.raw	Raw
	20221111_A375_clone5_CTRL_02_50ug.raw	Raw
	20221111_A375_clone5_CTRL_03_50ug.raw	Raw
	20221111_A375_clone5_shSEMA5A_02_50ug.raw	Raw
	20221111_A375_clone5_shSEMA5A_03_50ug.raw	Raw

Items per page:

1 - 5 of 7

Publications

Semaphorin 5A modulates focal adhesion pathway and lamellipodia formation in melanoma.

Brignone Matteo M Cufaro Maria Concetta MC Ercolani Cristiana C Masi Ilenia I Di Ferdinando Fabio F Palange Aldo A Valentini Elisabetta E Di Martile Marta M Del Boccio Piero P Del Bufalo Donatella D Tamagnone Luca L Rosanò Laura L D'Aguanno Simona S

Cell communication and signaling : CCS 20251127 1

<h4>Introduction</h4>Metastatic potential and rising incidence make cutaneous melanoma a major health concern worldwide. While current therapies have proven effective, they are limited by side effects arising and by the development of drug resistance. For this reason, identifying new therapeutic targets and predictive biomarkers is crucial. Semaphorin 5 A (SEMA5A) is a member of the Semaphorin family, implicated in both cancer biology and neural development. We have previously demonstrated that ...[more]

PMID: 41310710

Similar Datasets

Project description:Purpose: Core 3 derived glycans, a major type of O-glycan expressed by normal epithelial cells of the gastrointestinal tract, are downregulated during malignancy, because of loss of expression of functional β3-N-acetylglucosaminyltransferase-6 (core 3 synthase). We investigated the expression of core 3 synthase in normal pancreas and pancreatic cancer and evaluated the biological effects of re-expressing core 3 synthase in pancreatic cancer cells that had lost expression. Experimental Design: We determined that pancreatic tumors and tumor cell lines have lost expression of core 3 synthase. We therefore re-expressed in human pancreatic cancer cells (Capan-2 and FG) to investigate the contribution of core 3 glycans to malignant progression. Results: Pancreatic cancer cells expressing core 3 synthase showed reduced in vitro cell proliferation, migration and invasion compared with vector control cells. Expression of core 3 O-glycans induced altered expression of β1 integrin, decreased activation of focal adhesion kinase, led to the down regulation of expression of several genes including REG1α and FGFR3, and altered lamellipodia formation. The addition of a GlcNAc residue by core 3 synthase leads to the extension of the tumor associated Tn structure on MUC1. Orthotopic injection of FG cells expressing core 3 synthase into the pancreas of nude mice produced significantly smaller tumors and decreased metastasis to the surrounding tissues compared to vector control FG cells. Conclusions: These findings indicate that expression of core 3 derived O-glycans in pancreatic cancer cells suppresses tumor growth and metastasis through modulation of glycosylation of mucins and other cell surface and extracellular matrix proteins. Two-condition experiment, Core 3 synthase stable expression (C3) vs. vector control (PLVX) cells. Biological replicates: 3 Core 3 synthase stable expression, 3 vector control, independently grown and harvested. One replicate per array.