hirsutanol-a and Liver-Neoplasms

To investigate the effect of Nrf2 gene knockdown on hirsutanols A-induced cytotoxicity in cancer cells.. The changes in the cell viability following treatment with different concentrations of hirsutanols A was detected by MTT assay, and the generation of reactive oxygen species (ROS) was assayed using flow cytometry. AnnexinV-FITC apoptosis kit was used to detect the cell apoptosis. Nrf2 protein expression in HepG2 and SW480 cells transfected with the siRNA targeting Nrf2 was analyzed with Western blotting.. At the concentrations of 1.25, 2.5, 5, 10, 20 and 40 µmol/L, hirsutanols A obviously inhibited the cell proliferation of human liver cancer HepG2 and colon cancer SW480 cells in a concentration-dependent manner. The levels of hydrogen peroxide increased rapidly after hirsutanols A treatment in both HepG2 (30 µmol/L) and SW480 (15 µmol/L) cells. Hirsutanols A also induced apoptosis of the two cells. Pretreatment with 5 mmol/L NAC totally inhibited apoptosis and ROS accumulation in the two cells induced by hirsutanols A. Transfection of HepG2 and SW480 cells with the siRNA caused a significant reduction in Nrf2 protein expression, which resulted in an increased sensitivity of the cells to hirsutanols A.. Hirsutanols A can induce apoptosis in HepG2 and SW480 cells by promoting ROS production and up-regulating Nrf2 expression. Nrf2 knockdown by siRNA can increase the sensitivity of the cancer cells to hirsutanols A in vitro.

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Liver Neoplasms; NF-E2-Related Factor 2; Reactive Oxygen Species; RNA, Small Interfering; Sesquiterpenes

Hirsutanol A is a novel sesquiterpene compound purified from fungus chondrostereum sp in Sarcophyton tortuosum. Its pharmacologic effect has not been reported yet. This study aimed to investigate cytotoxic effect of Hirsutanol A on hepatocellular carcinoma (HCC) cells and its mechanism.. Hep3B cells were treated with different concentrations of Hirsutanol A. Cell proliferation was detected by MTT assay. The protein expression of LC3 was determined by Western blot. The generation of reactive oxygen species (ROS) was monitored by flow cytometry.. Hirsutanol A significantly inhibited proliferation of Hep3B cells with 50% inhibition concentrations (IC50) of 14.54, 6.71, and 3.59 micromol/L when exposed to Hirsutanol A for 24, 48, and 72 h, respectively. Incubation of Hep3B cells with Hirsutanol A markedly increased the level of ROS and the autophagy marker MAP-LC3 conversion from type I to type II. Pre-incubation with an antioxidant N-acetyl cystein (NAC) decreased the level of ROS, and reduced MAP-LC3 I-II conversion, and suppressed cell death. Blocking autophagy with a specific autophagy inhibitor 3-methyladenine (3-MA), the cytotoxic effect of this compound was attenuated.. Hirsutanol A has potent cytotoxic effect, and can induce autophagic cell death via increasing ROS production.

Topics: Acetylcysteine; Adenine; Agaricales; Antineoplastic Agents; Autophagy; Carcinoma, Hepatocellular; Cell Death; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Free Radical Scavengers; Humans; Liver Neoplasms; Microtubule-Associated Proteins; Reactive Oxygen Species; Sesquiterpenes