This confirms SPRED1 specifically suppresses Ras/MAPK signaling via MEK/ERK inhibition in HCC cells.​ Colony formation assays demonstrated that SPRED1 overexpression significantly inhibited Hep3B cell proliferation, and this effect was reversed by the Ras/MAPK agonist ML-098 (Fig. 3I), further supporting SPRED1's role in inhibiting HCC proliferation via Ras/MAPK suppression. Here, SPRED1 is linked to hepatocellular carcinoma.