Ophiopogonin-D significantly inhibited the in vitro and in vivo growth of prostate cells via RIPK1. Ophiopogonin-D may be developed as a potential anti-prostate cancer agent.

Ophiopogonin D(OPD) 对PC3细胞展现了强大的抗肿瘤活性。通过RIPK1相关途径诱导细胞凋亡，提高了RIPK1和Bim的蛋白表达水平，降低了cleaved-RIPK1、caspase 8、cleaved-caspase 8、Bid、caspase 10、及cleaved-caspase 10的水平。OPD还增加了Bim的mRNA表达。当细胞预先用necrostatin-1[1]处理时，Bim的蛋白表达水平降低。

Ophiopogonin D(OPD)′ 以5mg/kg剂量治疗，自治疗第6天起显著抑制肿瘤生长（p = 0.034）。研究结束时（第24天），切除、拍照并称重肿瘤组织。以5mg/kg OPD′治疗结果在第24天与对照治疗相比，肿瘤生长抑制显著（p = 0.000），约79.8%。5.0 mg/kg剂量的肿瘤生长抑制作用明显强于2.5 mg/kg剂量（p = 0.000）。所有组别均未发现显著的体重减轻[1]。

The CCK-8 assay was used to assess the viability of prostate cancer cells.?The cell morphology was examined by an ultrastructural analysis via transmission electron microscopy.?Cells in apoptosis (early and late stages) were detected using an Annexin V-FITC/propidium iodide kit with a FACSCaliber flow cytometer.?JC-1, a cationic lipophilic probe, was employed to measure the mitochondrial membrane potential (MMP) of PC3 cells.?Changes in the protein expression of RIPK1, C-RIPK1, caspase 8, cleaved-caspase 8, Bim, Bid, caspase 10, and cleaved-caspase 10 were evaluated by Western blotting.?The mRNA expression of Bim was examined by quantitative real-time reverse transcription polymerase chain reaction.?Z-VAD-FMK (a caspase inhibitor) and necrostatin-1 (a specific inhibitor of RIPK1) were utilized to determine whether the cell death was mediated by RIPK1 or caspases[1].

One week after tumor cell inoculation, mice bearing palpable tumors were randomly divided into control and treatment groups (8 mice/group).?OPD′ was dissolved in the vehicle, PEG400:Saline:Ethanol (400:300:200, v/v/v), and administered (via i.p. injection) at doses of 2.5 or 5.0 mg/kg bodyweight 5 days a week for 24 days.?The control group received vehicle only.?The mice were sacrificed by cervical dislocation on Day 24, and the tumor tissues were removed and weighed[1].