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Endoplasmic reticulum stress induced by an ethanol extract of Coicis semen in Chang liver cells.

ABSTRACT: It is well known that endoplasmic reticulum (ER) stress plays a huge role in development of metabolic diseases. Specially, ER stress-induced cellular dysfunction has a significant involvement in the pathogenesis of human chronic disorders. This study was designed to study to assess whether an ethanol extract of Coicis Semen (CSE) and coixol induces the ER stress in Chang liver cells.Coicis Semen was mixed with 95% ethanol at a ratio of 1:10 (w/v) and freeze dried. Chang liver cells were seeded to 96-well plates and treated with or without CSE (100, 200, 300, 500, or 1000 ?g/mL) or coixol (100, 200, 300, 500, 750, or 1000 ?g/mL). cell viability was analyzed with MTT assay. Effects of CSE and coixol on expression of the genes for ER stress markers were determined with qRT-PCR and the expression of the protein levels of ER stress markers were determined with western blotting.The concentration causing 50% inhibition (IC50) for CSE and coixol was 250 and 350 ?g/mL, respectively. The CSE and coixol increased the gene expression of BiP and CHOP in a dose-dependent manner. Furthermore, CSE and coixol dose-dependently increased the the expression of XBP1.CSE or coixol may have cytotoxic effect to Chang liver cells and, may induce ER stress and stimulate the UPR via activation of the PERK and IRE1 pathways in normal liver cells.

SUBMITTER: Kim HY

PROVIDER: S-EPMC5859727 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Endoplasmic reticulum stress induced by an ethanol extract of Coicis semen in Chang liver cells.

Kim Hwa Yeon HY Song Ha Na HN Davaatseren Munkhtugs M Chang Hyun Joo HJ Chun Hyang Sook HS

BMC complementary and alternative medicine 20180320 1

<h4>Background</h4>It is well known that endoplasmic reticulum (ER) stress plays a huge role in development of metabolic diseases. Specially, ER stress-induced cellular dysfunction has a significant involvement in the pathogenesis of human chronic disorders. This study was designed to study to assess whether an ethanol extract of Coicis Semen (CSE) and coixol induces the ER stress in Chang liver cells.<h4>Methods</h4>Coicis Semen was mixed with 95% ethanol at a ratio of 1:10 (w/v) and freeze dri ...[more]

PMID: 29554897

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Project description:BackgroundMarchantia polymorpha L. is a kind of Chinese herbal medicine and has various biological activities including antioxidant and antifungal. However, it is not clear about the antitumor effect and mechanism of M. polymorpha. We prepared M. polymorpha ethanol extract (MPEE) and investigated its antitumor effect on hepatocellular carcinoma cells both in vitro and in vivo.MethodsThe viability of hepatocellular carcinoma cells was detected by MTT assay. The distribution of cell cycle was analyzed by propidium iodide (PI) staining. The morphology of nuclei was observed by Hoechst 33258 staining. Apoptosis was detected by Annexin V/PI staining. JC-1 fluorescent probe and DCFH-DA were used to detect the mitochondrial membrane potential (ΔψM) and the level of reactive oxygen species (ROS), respectively. Caspase inhibitors were used to test the function of caspase in the induction of apoptosis. Quantitative real time polymerase chain reaction (qRT-PCR) and Western blot were used to evaluate the levels of mRNA and protein, respectively. Differentially expressed genes and signaling pathways were identified by transcriptome analysis. The H22 tumor mouse model was used to detect the antitumor effect of the extract.ResultsMPEE significantly suppressed the migration and growth of BEL-7404, HepG2 and H22 cells in a dose- and time-dependent manner through induction of apoptosis characterized by chromosomal condensation and cell cycle arrest at G0/G1 and G2/M phases. MPEE induced mitochondria-dependent apoptosis via upregulation of Bax and downregulation of Bcl-2 to reduce mitochondrial membrane potential and increase the release of cytochrome c. The levels of cleaved caspase-8 and -9 were significantly increased, which sequentially activated caspase-3 to cleave PARP. We further found that MPEE significantly increased ROS production and activated endoplasmic reticulum (ER) stress associated-apoptotic signaling pathway. Moreover, MPEE significantly inhibited H22 tumor growth in mouse model and improved the survival of tumor mice.ConclusionThese results suggested that MPEE suppressed hepatocellular carcinoma cell growth through induction of apoptosis via intrinsic- and ER stress-associated pathways.

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Endoplasmic reticulum stress induced by an ethanol extract of Coicis semen in Chang liver cells.

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Endoplasmic reticulum stress induced by an ethanol extract of Coicis semen in Chang liver cells.

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