OBJECTIVETo investigate the effects of PsL5F (ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid, an extract from Pteris semipinnata L) on the expression of nuclear receptor subfamily 1, group D, member 1 (Nr1d1) in highly metastatic ovarian carcinoma HO-8910PM cells, and its mechanisms.

METHODMicroarray Chip was used to examine the level of Nr1d1 mRNA expression on HO-8910PM cells treated with PsL5F. Fluorescent quantitative real-time PCR assay and Western blot were performed to verify the effects of PsL5F on Nr1d1 mRNA and protein expression.

RESULTAfter 24 h treatment of 100 micromol x L(-1) PsL5F, the mRNA and protein levels of Nr1d1 in HO-8910PM cells were 35.34 +/- 1.07 and 7.71 +/- 0.43 times compared to those of control group (P < 0.01, P < 0.01), respectively.

CONCLUSIONPsL5F can up-regulate significantly the expression of Nr1d1 in HO-8910PM cells. Antitumor effects and its mechanisms of PsL5F in HO-8910PM cells may be involved in the up-regulation of Nr1d1 expression.

Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Female ; Gene Expression ; drug effects ; Humans ; Neoplasm Invasiveness ; Ovarian Neoplasms ; pathology ; Piperidones ; pharmacology ; Pteris ; chemistry ; RNA, Messenger ; drug effects ; metabolism

OBJECTIVETo identify the role of reactive oxygen species (ROS) formation on cell death induced by Ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F) in HepG2 cells.

METHODMTT assay was used to determine the effect of 5F on proliferation of HepG2 cells, and apoptotic morphological changes were assessed using Hoechst/PI assay. To evaluate intracellular ROS levels, a GENMED kit was used. HepG2 cells were treated with 5F for 24 h or with 1 mmol x L(-1) GSH for 1 h prior to treatment with 5F for 24 h, then cytoplasmic mono- and oligonucleosomes were assessed with Cell Death Detection ELISA kit.

RESULTThe cytotoxicity of 5F on HepG2 cells was elevated with increasing 5F concentrations, as evidenced by the cell viability assay, and the apoptotic changes such as chromatin condensation were confirmed by Hoechst/PI staining. The decrease in ROS generation was observed in HepG2 cells following treatment with 5F. Cytoplasmic mono- and oligonucleosomes induced by 5F were not changed by decreasing basal level of ROS-mediated signaling with GSH. Further more, induction of ROS production by cisplatinum (CDDP) was canceled by treatment with 5F and 5F revealed a additive effect to cell killing by CDDP.

CONCLUSION5F can not only induce apoptosis through non-ROS-depandent pathway, and can abate oxidant stress.

Apoptosis ; drug effects ; Cell Death ; drug effects ; Diterpenes ; toxicity ; Drugs, Chinese Herbal ; toxicity ; Hep G2 Cells ; Humans ; Pteris ; chemistry ; Reactive Oxygen Species ; metabolism