Our study shows that, knockdown of both miR-221 and AEG-1 inhibits cell migration, invasion, proliferation, and angiogenesis, and enhances apoptosis and cell cycle arrest in sub G0-G1/G2-M phases by upregulating PTEN, p57, p53, and RB and downregulating LSF, MMP9, BCL2, PI3K, AKT, and LC3A genes in HCC cells. Here, MTDH is linked to hepatocellular carcinoma.