Research Paper Volume 11, Issue 21 pp 9719—9737

Tanshinone IIA mediates SMAD7-YAP interaction to inhibit liver cancer growth by inactivating the transforming growth factor beta signaling pathway

Figure 2. Tan IIA can regulate TGF-β/SMADs and Hippo/YAP signaling pathway in liver cancer cells. (A) Bel-7404 and SMMC-7721 cells were cultured with DMSO or dose dependent Tan IIA (5, 10, 20, 40 μM) for 24 h and protein expression levels of Ki67, CyclinD1, Bcl2, Cleaved Caspase Substrate, E-cadherin and N-cadherin were measured by western blot assay. (B) GO biological process and KEGG pathway enrichment of SMAD7 co-expressed top 50 genes from cBioPortal database. (C) TGF-β/SMADs and Hippo/YAP signaling pathway related genes were determined by real-time quantitative PCR assay with DMSO or Tan IIA (40 μM). (D) The protein expression level of TGF-β/SMADs components TGFβR1, SMAD2, SMAD3, P-SMAD2, SMAD7 and Hippo pathway effector YAP and E3 ligase SMURF1, βTrcp were analyzed by western blot assay with DMSO, 20 μM, 40 μM Tan IIA in Bel-7404 cells. (E) Indicated protein expression levels were measured by western blot assay with or without SMAD7-Flag overexpression in Bel-7404 and SMMC-7721 cells.