Fig. 5

The metalloproteinase activity of ADAMTS1 is pivotal in stimulating the secretion of cleaved versican, activating the epidermal growth factor receptor (EGFR), promoting invasion, and conferring anoikis resistance in renal cell carcinoma (RCC) cells. A A schematic representation depicting two variants of the recombinant ADAMTS1 protein: one containing the metalloproteinase domain (ZnMc) and the other lacking it (TSP), along with a mutant ADAMTS1 expression construct (E402Q). B–E Treatment of Caki-1 cells with or without the recombinant ZnMc or TSP protein (40 nM) for 24 or 48 h. Subsequently, secretion of cleaved versican, activation of EGFR signal cascades, invasive abilities, and anoikis were respectively assessed using dot blot (B), western blotting (C), Matrigel invasion (D), and CCK8 assays. F–I Cleaved versican secretion (F), EGFR activation (G), invasive abilities (H), and anoikis (I) were evaluated in Caki-1 and 786-O cells after transducing wild-type (WT) ADAMTS1, ADAMTS1/E402Q, or a control vector. In D, E, H, and I, values are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, compared with the control group; ^#p < 0.05, ^###p < 0.001, compared with the WT ADAMTS1-overexpressing group