title: Goniothalamin induces mitochondria-mediated apoptosis associated with endoplasmic reticulum stress-induced activation of JNK in HeLa cells.
abstract: Goniothalamin, a natural occurring styryl-lactone isolated fromGoniothalamus macrophyllus(Blume) Hook. f. & Thomson var.macrophyllus, can trigger cancer cell death in various types of cancer cell. The present study focused on elucidation of the mitochondria-mediated apoptosis associated with endoplasmic reticulum (ER) stress-induced activation of c-Jun NH2-terminal kinase (JNK) by goniothalamin in HeLa cervical cancer cells. Cell viability was determined using an MTT assay, and DNA condensation and loss of mitochondrial membrane potential were determined using Hoechst 33342 and JC-1 staining, respectively. Flow cytometry was used for cell cycle and phosphatidyl-serine exposure analyses. Apoptotic-associated ER stress signaling pathways were determined using immunoblotting, reverse transcription-polymerase chain reaction (RT-PCR) and RT-quantitative PCR analyses. The results suggested that goniothalamin suppressed cell proliferation in a time- and dose-dependent manner. The induction of apoptosis was confirmed by increased DNA condensation, loss of mitochondrial membrane potential and cell surface phosphatidyl-serine presentation. The cell cycle analysis demonstrated that the goniothalamin-treated HeLa cells were in G2/M arrest. Determination of the caspase cascade and apoptotic proteins indicated the induction of apoptosis through the intrinsic pathway. In addition, the levels of phosphorylated JNK and the transcription factor, C/EBP homologous protein (CHOP), an ER stress-associated apoptotic molecule, were increased in the goniothalamin-treated cells. These data indicated that goniothalamin exerted a cytotoxic effect against HeLa cells via the induction of mitochondria-mediated apoptosis, associated with ER stress-induced activation of JNK.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Srinkharinwirot University, Bangkok 10110, Thailand.
authors.forename: Thanet
authors.initials: T
authors.lastname: Sophonnithiprasert
authors.affiliation: Department of Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand.
authors.forename: Wilawan
authors.initials: W
authors.lastname: Mahabusarakam
authors.affiliation: Department of Orthopaedic Surgery, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Srinkharinwirot University, Bangkok 10110, Thailand.
authors.forename: Ramida
authors.initials: R
authors.lastname: Watanapokasin
keywords: apoptosis; c-Jun NH2-terminal kinase; cervical cancer; endoplasmic reticulum stress; goniothalamin; transcription factor C/EBP homologous protein