桂枝茯苓丸调控MTDH和PTEN逆转卵巢癌多药耐药的机制研究

研究桂枝茯苓丸在人卵巢癌耐药性裸鼠肿瘤模型中对凋亡的诱导作用,探讨其可能的作用机制。采用实时无标记细胞分析系统检测SKOV3/DDP对顺铂和紫杉醇的耐药性,建立SKOV3/DDP卵巢癌耐药性BALB/c裸鼠肿瘤模型,模型裸鼠随机分为桂枝茯苓丸浓缩液16 g.kg-1.d-1(高)、8 g.kg-1.d-1(中)、4 g.kg-1.d-1(低)剂量组、PI3K抑制剂LY294002组、低剂量桂枝茯苓丸LY294002组(联用组)及对照组,给药10天后剖取肿瘤组织,采用流式细胞术检测肿瘤细胞的凋亡率,采用荧光定量PCR检测肿瘤组织中MTDH、PTEN mRNA的相对表达。结果表明，SKOV3/DDP细胞对顺铂和紫杉醇的耐药倍数分别为4.57和4.36倍。随着桂枝茯苓丸浓缩液剂量的增加,肿瘤细胞的凋亡率逐渐增加,上调PTEN mRNA和下调MTDH mRNA的作用逐渐增强,各组之间差异具有显著性(P < 0.01)。LY294002和低剂量桂枝茯苓丸联用后,抑制凋亡的作用强于二者分别单用的凋亡抑制作用,下调MTDH mRNA和上调PTEN mRNA的作用强于其他各组(P < 0.01)。说明在SKOV3/DDP卵巢癌耐药性BALB/c裸鼠模型中,桂枝茯苓丸能够剂量依赖性地诱导耐药性肿瘤细胞的凋亡,其机制可能与上调PTEN mRNA和下调MTDH mRNA有关。

Mechanism of Guizhi Fuling Wan in Regulating MTDH and PTEN to Reverse Multidrug Resistance of Ovarian Cancer

To study the apoptosis inducing effect of Guizhi Fuling Wan (GFW) in human ovarian cancer-resistant nude mouse tumor model, and to explore the possible mechanism. The resistance of SKOV3/DDP cells to paclitaxel or cisplatin was tested by a real-time cell analyzer. A resistant ovarian cancer BALB/c xenograft nude mice model were established. The model mice were randomly divided into six groups: GFW low (4 g.kg-1.d-1), middle (8 g.kg-1.d-1) and high (16 g.kg-1.d-1) doses groups, PI3K inhibitor LY294002 group, LY294002 combined with low dose of GFW group (Combined group) and control group. Tumor tissues were dissected after 10 days administration, the apoptosis rates were analyzed by flow cytometry in the different groups. The relative changes of PTEN and MTDH mRNA in tumor tissues were examined by a real-time quantitative PCR. The results showed that the resistance times of SKOV3/DDP cells to paclitaxel or cisplatin were 4.57 and 4.36, respectively. The apoptosis rate was increased with the dose of GFW increased (P < 0.01). A similar effect was observed for the relative changes of the PTEN and MTDH mRNA level in tumor tissues compared with that in control group (P < 0.01). After LY294002 combined with low dose of GFW, the effect of inhibiting apoptosis was stronger than that of the two groups alone, the effect of down-regulating MTDH mRNA and up-regulating PTEN mRNA was stronger than that in other groups (P < 0.01). In conclusion, GFW can induce apoptosis of drug-resistant tumor cells in a dose-dependent manner, which may be related to the up-regulation of PTEN mRNA and down-regulation of MTDH mRNA in SKOV3/DDP ovarian cancer-resistant BALB/c nude mice models.