Lu Zongliang, Wang He, Zhu Mingxing, Song Wei, Wang Jiajia, Wu Changpeng, Kong Ya, Guo Jing, Li Na, Liu Jie, Li Yanwu, Xu Hongxia
Department of Nutrition, Daping Hospital and Research Institute of Surgery, Third Military Medical University, Chongqing, China.
Department of Clinical Nutrition, Yubei District People's Hospital, Chongqing, China.
Front Pharmacol. 2018 Apr 30;9:432. doi: 10.3389/fphar.2018.00432. eCollection 2018.
The purpose of this study was to evaluate the anticancer effects of Ophiopogonin D' (OPD', a natural product extracted from a traditional Chinese medicine () against androgen-independent prostate cancer cells and to explore the underlying molecular mechanism(s) of action. 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. PC3 and DU145 xenograft models in BALB/c nude mice were used to evaluate the anticancer activity of OPD' . OPD' was shown to exert potent anti-tumor activity against PC3 cells. It induced apoptosis via a RIPK1-related pathway, increased the protein expression levels of RIPK1 and Bim, and decreased the levels of cleaved-RIPK1, caspase 8, cleaved-caspase 8, Bid, caspase 10, and cleaved-caspase 10. OPD' also increased the mRNA expression of Bim. The protein expression of Bim was decreased when cells were pre-treated with necrostatin-1. Treatment with OPD' inhibited the growth of PC3 and DU145 xenograft tumors in BALB/c nude mice. OPD' significantly inhibited the and growth of prostate cells via RIPK1, suggesting that OPD' may be developed as a potential anti-prostate cancer agent.
本研究的目的是评估麦冬皂苷D'（OPD'，一种从中药中提取的天然产物）对雄激素非依赖性前列腺癌细胞的抗癌作用，并探索其潜在的分子作用机制。采用CCK-8法评估前列腺癌细胞的活力。通过透射电子显微镜进行超微结构分析来检测细胞形态。使用Annexin V-FITC/碘化丙啶试剂盒和FACSCaliber流式细胞仪检测凋亡（早期和晚期）细胞。采用阳离子亲脂性探针JC-1来测量PC3细胞的线粒体膜电位（MMP）。通过蛋白质印迹法评估RIPK1、C-RIPK1、半胱天冬酶8、裂解的半胱天冬酶8、Bim、Bid、半胱天冬酶10和裂解的半胱天冬酶10的蛋白表达变化。通过定量实时逆转录聚合酶链反应检测Bim的mRNA表达。使用Z-VAD-FMK（一种半胱天冬酶抑制剂）和坏死抑制剂-1（RIPK1的特异性抑制剂）来确定细胞死亡是否由RIPK1或半胱天冬酶介导。利用BALB/c裸鼠中的PC3和DU145异种移植模型来评估OPD'的抗癌活性。结果显示，OPD'对PC3细胞具有强大的抗肿瘤活性。它通过RIPK1相关途径诱导细胞凋亡，增加RIPK1和Bim的蛋白表达水平，并降低裂解的RIPK1、半胱天冬酶8、裂解的半胱天冬酶8、Bid、半胱天冬酶10和裂解的半胱天冬酶10的水平。OPD'还增加了Bim的mRNA表达。当细胞用坏死抑制剂-1预处理时，Bim的蛋白表达降低。用OPD'处理可抑制BALB/c裸鼠中PC3和DU145异种移植瘤的生长。OPD'通过RIPK1显著抑制前列腺细胞的生长，表明OPD'可能被开发成为一种潜在的抗前列腺癌药物。
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