Theaflavin 3,3′-digallate suppresses metastasis and reduces insulin-like growth factor-1-induced cancer stemness and invasiveness in human melanoma cells

Theaflavin 3,3′-digallate (TF3), a flavonoid compound, exerts antidiabetic effects, induces apoptosis, and suppresses cancer growth. However, its effects on melanoma metastasis and insulin-like growth factor 1 (IGF-1)-induced cancer stemness remain largely unexplored. This study explored key molecular signaling pathways underlying the antitumor and antimetastatic activities of TF3 in melanoma. The antimetastatic potential of TF3, as well as its impact on IGF-1-induced cancer stemness, were evaluated using wound healing assays, gelatin zymography, Matrigel invasion assays, sphere formation assays, and Western blotting. Its antiangiogenic and antimetastatic activities were further examined in vivo using subcutaneous xenograft and tail vein injection mouse models. TF3 significantly inhibited cell migration, invasion, and matrix metalloproteinase (MMP) activity in A 375 and A2058 melanoma cells. It also suppressed sphere formation, self-renewal capacity, and aldehyde dehydrogenase 1 (ALDH1) activity. In sphere-forming melanoma cells, TF3 downregulated key cancer stemness and drug resistance markers, including ABCB1, ABCG2, CD44, and CXCR4. Notably, TF3 reversed IGF-1-induced sphere formation, invasion, and MMP-9 expression. Mechanistically, TF3 suppressed IGF-1-mediated phosphorylation of focal adhesion kinase (FAK) and Src. In vivo, TF3 significantly inhibited tumor growth, reduced angiogenic marker expression, and suppressed lung metastasis. Collectively, these findings demonstrate that TF3 impedes melanoma progression by inhibiting metastasis and suppressing IGF-1-induced cancer stemness and invasiveness.