title: Endoplasmic reticulum stress mediates sulforaphane-induced apoptosis of HepG2 human hepatocellular carcinoma cells.
abstract: Sulforaphane (SFN) is a naturally occurring chemopreventive agent, which effectively inhibits proliferation of HepG2 human hepatocellular carcinoma cells via mitochondria‑mediated apoptosis. Endoplasmic reticulum stress is considered the most important cause of cell apoptosis; therefore, the present study aimed to determine whether the endoplasmic reticulum pathway was involved in SFN-induced apoptosis of HepG2 cells. An MTT assay was used to detect the inhibitory effects of SFN on HepG2 cells. Fluorescence microscopy was used to observe the morphological changes in apoptotic cells, and western blot analysis was conducted to detect the expression of binding immunoglobulin protein (Bip)/glucose-regulated protein 78 (GRP78), X‑box binding protein‑1 (XBP‑1) and BH3 interacting domain death agonist (Bid). Furthermore, flow cytometry was used to determine the apoptotic rate of HepG2 cells, and the protein expression of C/EBP homologous protein (CHOP)/growth arrest‑ and DNA damage‑inducible gene 153 (GADD153) and caspase-12 in HepG2 cells. The results indicated that SFN significantly inhibited the proliferation of HepG2 cells; the half maximal inhibitory concentration values were 32.03±0.96, 20.90±1.96 and 13.87±0.44 µmol/l, following treatment with SFN for 24, 48 and 72 h, respectively. Following 48 h of SFN treatment (10, 20 and 40 µmol/l), the apoptotic rates of HepG2 cells were 31.8, 61.3 and 77.1%, respectively. Furthermore, after 48 h of exposure to SFN, the cells presented typical morphological alterations of apoptosis, as detected under fluorescence microscopy. Treatment with SFN for 48 h also significantly upregulated the protein expression levels of Bip/GRP78, XBP‑1, caspase‑12, CHOP/GADD153 and Bid in HepG2 cells. In conclusion, endoplasmic reticulum stress may be considered the most important mechanism underlying SFN-induced apoptosis in HepG2 cells.
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Ministry of Education, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zou
authors.affiliation: College of Pharmacy, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Zhongyuan
authors.initials: Z
authors.lastname: Qu
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Ministry of Education, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Yueni
authors.initials: Y
authors.lastname: Fang
authors.affiliation: Post Doctoral Research Center of Materia Medica, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Shi
authors.affiliation: Post Doctoral Research Center of Materia Medica, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Yubin
authors.initials: Y
authors.lastname: Ji
mesh_terms: D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D053454:Caspase 12 / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D006360:Heat-Shock Proteins / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D017879:Isothiocyanates / Q000494:pharmacology*; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D013454:Sulfoxides; D051742:Transcription Factor CHOP / Q000378:metabolism; D000071717:X-Box Binding Protein 1 / Q000378:metabolism
chemical_list: D016588:Anticarcinogenic Agents; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; C497249:DDIT3 protein, human; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D017879:Isothiocyanates; D013454:Sulfoxides; D000071717:X-Box Binding Protein 1; D051742:Transcription Factor CHOP; D053454:Caspase 12; C016766:sulforaphane