Ovarian and Prostate Cancer Research Trust Laboratory, Guildford, Surrey GU2 7AF, UK.
Cell Biol Toxicol. 2011 Apr;27(2):133-47. doi: 10.1007/s10565-010-9176-0. Epub 2010 Sep 26.
The initial response rates of advanced-stage epithelial ovarian cancer to the chemotherapeutic agents carboplatin and paclitaxel are high. However, once drug resistance develops, further chemotherapy is less effective. The objective of this study is to investigate the anti-proliferative activity of the phyto-active chemicals (PACs) oridonin and wogonin in chemo-resistant epithelial ovarian cancer cells. Primary cell cultures from the ascitic fluid of three patients at diagnosis, two patients chemo-resistant to carboplatin and paclitaxel, and one patient treated with letrozole for breast cancer were studied and compared to the ovarian cancer cell lines A2780 and PTX10, by cell viability assay (MTS). Effects on cell cycle modulation and apoptosis were examined by flow cytometry and Western blot analysis (WB). WB was further conducted to investigate protein expressions altered by PACs. The results show that IC(50) of the primary cultures ranged from 0.6 to 5.4 μg/ml for oridonin and 0.3-12.7 μg/ml for wogonin. The paclitaxel-resistant cell line PTX10 was more sensitive to each of the PACs than the chemo-sensitive cell line A2780. Of particular interest is that in combination, the two PACs were synergistic in their cytotoxicity to five of six of the primary cultures and to both the cell lines (combination indices of 0.39-0.95). The inhibition is attributable to apoptosis and cell cycle modulation induced by the PACs as demonstrated in A2780 and PTX10. Up-regulation of the functional p53 protein in A2780 and down-regulation of Akt protein in PTX10 have in part contributed to the apoptosis. These findings suggest that oridonin and wogonin may have activity in ovarian cancer following its development of resistance to carboplatin and paclitaxel.
晚期上皮性卵巢癌对化疗药物卡铂和紫杉醇的初始缓解率较高。然而，一旦产生耐药性，进一步的化疗效果就会降低。本研究的目的是研究植物活性化学物质（PACs）冬凌草甲素和汉黄芩素在耐药性上皮性卵巢癌细胞中的抗增殖活性。从 3 名患者的腹水中分离原代细胞，其中 2 名患者对卡铂和紫杉醇耐药，1 名患者接受来曲唑治疗乳腺癌，将其与卵巢癌细胞系 A2780 和 PTX10 进行比较，通过细胞活力测定（MTS）进行研究。通过流式细胞术和 Western blot 分析（WB）检测细胞周期调节和细胞凋亡的影响。WB 进一步用于研究 PACs 改变的蛋白表达。结果表明，原代培养物中冬凌草甲素的 IC50 范围为 0.6-5.4μg/ml，汉黄芩素的 IC50 范围为 0.3-12.7μg/ml。紫杉醇耐药细胞系 PTX10 对每种 PAC 的敏感性均高于化疗敏感细胞系 A2780。特别值得注意的是，在组合中，两种 PAC 对六种原代培养物中的五种和两种细胞系（组合指数为 0.39-0.95）具有协同细胞毒性。这种抑制归因于 PAC 诱导的细胞凋亡和细胞周期调节，这在 A2780 和 PTX10 中得到了证明。A2780 中功能性 p53 蛋白的上调和 PTX10 中 Akt 蛋白的下调部分促成了细胞凋亡。这些发现表明，冬凌草甲素和汉黄芩素可能在卵巢癌对卡铂和紫杉醇产生耐药性后具有活性。
