Compound Astragalus and Salvia miltiorrhiza extract exerts anti-tumor activity by intervening the interaction of microRNA-145/microRNA-21 and Smad3C/3L phos-phorylation in hepatocellular carcinoma / 中国药理学与毒理学杂志

ABSTRACT

OBJECTIVBE To investigate the intervention of compound Astragalus and Salvia milt-iorrhiza extract (CASE) consisted of astragalosides, astragalus polysaccharides and salvianolic acids on the interaction of microRNA-145/microRNA-21 (miR-145/miR-21) and Smad3C/3L phosphorylation (pSmad3C/pSmad3L) down-stream of transforming growth factor-β (TGF-β)/mitogen activated protein kinase (MAPK) signaling in hepatocellular carcinoma (HCC) progression by in vitro and in vivo experi-ments. METHODS In HepG2 cells and xenografts of nude mice, antagomir/agomir and plasmids of Smad3C/3L phosphorylation site mutation (Smad3 3S-A/Smad3 EPSM) were used to intervene miR-145/miR-21 and pSmad3C/pSmad3L expression respectively,then incorporative CASE treatment. Cell proliferation, migration, apoptosis, tumor growth and histopathologic characteristics of xenografts, relevant proteins of TGF-β/Smad pathway and miR-145/miR-21 were evaluated.RESULTS CASE up-regulated miR-145 while down-regulated miR-21, inhibited cell proliferation,migration and tumor growth, accelerated cell apoptosis in HepG2 cells respectively transfected with Smad3 WT, Smad3 EPSM,Smad3 3S-A plasmids in cultured dishes and xenografts of nude mice,the above effects were more evident in HepG2 cells with increased pSmad3C.In TGF-β1-stimulated HepG2 cells and xenografts of nude mice, CASE antagonized the facilitating effects of miR-145 antagomir/miR-21 agomir on cell migration,proliferation,tumor growth and inhibiting effects of miR-145 antagomir/miR-21 agomir on cell apoptosis; CASE increased miR-145 down-regulated by miR-145 antagomir and decreased miR-21 up-regulated by miR-21 agomir,reduced protein level of pSmad3L and their proteins including TβRⅡ, pERK1/2, pJNK1/2 and pp38 while elevated pSmad3C expression. CONCLUSION These results suggest that pSmad3C/pSmad3L maybe interact with miR-145/miR-21 in HCC progression,which may be one of important molecular mechanisms of CASE's anti-HCC effects.