Cancer Research Institute, Zhejiang Cancer Hospital, Hangzhou 310022, China.
J Tradit Chin Med. 2013 Jun;33(3):325-33. doi: 10.1016/s0254-6272(13)60174-0.
To study the anticancer mechanism of polyphyllin I (PPI), a Traditional Chinese Medicine, on the ovarian cancer cell line HO-8910PM in vitro.
Transwell chamber invasive assays were used to investigate the inhibitory capacity of PPI on HO-8910PM metastasis. Gene expression profiling chips was used to screen differentially expressed genes between experiment group and control group. Reverse transcription PCR and Western blotting were used to determine mRNA and protein levels.
With increasing PPI concentration, the metastatic capacity of cells decreased, with significance differences between the experimental and control groups (P < 0.01) as well as between two concentration groups. Gene expression profiling identified 123 differentially expressed genes, of which 70 were downregulated and 53 were upregulated. The genes were involved in multiple signal transduction pathways, including apoptosis, proliferation and metastasis. Real-time PCR (RT-PCR) showed that differential genes PIK3C2B, Caspase 9 and Wnt5A were downregulated with increasing PPI, showing an evident dose-effect relationship. The c-Jun was an exception. As the PPI dosage increased and the exposure time was extended, c-Jun relative expression showed an upward trend. There were significant differences between the experiment and control (P < 0.05). Western blot analyses showed that PPI treatment decreased levels of Caspase 9, Wnt5A and PIK3C2B and increased activated Caspase 9, c-Jun and p-c-Jun expression levels.
PPI has strong antitumor and anti transfer activity. It can activate c-Jun expression and the JNK signaling pathway, elicit cell apoptosis via the mitochondrial-mediated Caspase activation pathway, and finally inhibit tumor growth and migration in vitro. The downregulation of PIK3C2B and Wnt5A jointly inhibit the proliferation and metastasis of HO-8910PM. PPI may be a novel treatment for ovarian cancer.
研究中药重楼甾体总苷(PPI)在体外对卵巢癌细胞系 HO-8910PM 的抗癌机制。
采用 Transwell 室侵袭实验研究 PPI 对 HO-8910PM 转移的抑制作用。基因表达谱芯片筛选实验组与对照组之间差异表达的基因。逆转录 PCR 和 Western blot 法测定 mRNA 和蛋白水平。
随着 PPI 浓度的增加,细胞的转移能力下降,实验组与对照组之间差异有统计学意义(P < 0.01),两个浓度组之间差异亦有统计学意义。基因表达谱鉴定出 123 个差异表达基因,其中 70 个下调,53 个上调。这些基因参与了多个信号转导通路,包括凋亡、增殖和转移。实时 PCR(RT-PCR)显示,差异表达基因 PIK3C2B、Caspase 9 和 Wnt5A 随 PPI 增加而下调,呈明显的剂量效应关系。c-Jun 是个例外。随着 PPI 剂量的增加和暴露时间的延长,c-Jun 的相对表达呈上升趋势。实验组与对照组之间差异有统计学意义(P < 0.05)。Western blot 分析显示,PPI 处理降低了 Caspase 9、Wnt5A 和 PIK3C2B 的表达水平,增加了活化 Caspase 9、c-Jun 和 p-c-Jun 的表达水平。
PPI 具有较强的抗肿瘤和抗转移活性。它可以通过激活 JNK 信号通路激活 c-Jun 表达,通过线粒体介导的 Caspase 激活途径引发细胞凋亡,最终抑制体外肿瘤的生长和迁移。PIK3C2B 和 Wnt5A 的下调共同抑制 HO-8910PM 的增殖和转移。PPI 可能是治疗卵巢癌的一种新方法。