Chlorotoxin suppresses melanoma vasculogenic mimicry via LINC01235/miR-128-3p/YY1/ VEGFR2/NRP1 axis

Vasculogenic mimicry (VM), a process of vascular-like channel formation for nutrient acquisition, drives melanoma metastasis and poor prognosis. We demonstrate that Chlorotoxin (CTX) effectively targets VM in melanoma. In A375 and A2058 cells, CTX dose-dependently inhibits proliferation with IC50 values of 1.0 and 0.8 μM, respectively. Furthermore, sub-lethal CTX (0.4 μM) impairs cell migration and VM tube formation while downregulating VM regulators VEGFR2 and NRP1. Mechanistically, CTX suppresses the long non-coding RNA LINC01235, a competing endogenous RNA that sponges miR-128-3p. CTX-induced downregulation of LINC01235 frees miR-128-3p to target and suppress the transcription factor YY1. This reduction in YY1 diminishes its binding to the VEGFR2 promoter, ultimately decreasing VEGFR2 and NRP1 transcription. Rescue assays confirm that LINC01235 overexpression restores VM capacity. By defining the LINC01235/miR-128-3p/YY1/VEGFR2/NRP1 signaling axis disrupted by CTX, these findings establish the mechanistic basis for CTX as a promising therapeutic agent against aggressive, VM-competent melanoma.