冬凌草甲素通过上调miR-204-5p抑制PC-3细胞迁移与侵袭的作用机制研究

目的 探索冬凌草甲素影响PC-3细胞迁移及侵袭的作用及机制，为冬凌草甲素治疗前列腺癌骨转移提供前期体外实验参考。 方法 通过CCK8检测冬凌草甲素对PC-3细胞增殖的影响，Transwell检测冬凌草甲素对PC-3细胞迁移和侵袭的影响，qRT-PCR分析冬凌草甲素溶液处理PC-3细胞后miR-204-5p及NF-κB信号通路表达情况，Western blotting分析冬凌草甲素溶液处理PC-3细胞后NF-κB信号通路蛋白表达情况。 结果 冬凌草甲素溶液以浓度依赖性和时间依赖性方式抑制PC-3细胞的增殖，与对照组相比差异有统计学意义（P<0.01），冬凌草甲素IC50值为10 μmol/L；冬凌草甲素溶液呈浓度依赖性抑制PC-3细胞的迁移和侵袭，差异较对照组有统计学意义（P<0.01）；冬凌草甲素溶液能促进PC-3细胞表达miR-204-5p并阻断NF-κB信号通路下游基因表达；过表达miR-204-5p和冬凌草甲素均能抑制PC-3细胞NF-κB信号通路下游基因表达。 结论 冬凌草甲素能抑制PC-3细胞的增殖、迁移及侵袭，其作用机制可能与其能上调miR-204-5p阻断NF-κB信号通路有关。

Study on the mechanism of oridonin inhibiting PC-3 cells migration and invasion by up-regulating miR-204-5p

【Abstract】 Objective To explore the effect and mechanism of oridonin on the migration and invasion of PC-3 cells, to provide preliminary in vitro experiment reference for the treatment of bone metastasis of prostate cancer with oridonin. Methods The effect of oridonin on the proliferation of PC-3 cells was detected by CCK8. The effect of oridonin on the migration and invasion of PC-3 cells was detected by the Transwell test. The expression of miR-204-5p and NF-κB signaling pathways after the treatment of PC-3 cells with oridonin was analyzed by qRT-PCR. The expression of protein in the NF-κB signaling pathway after the treatment of PC-3 cells with oridonin was analyzed by Western blotting. Results Firstly, the proliferation of PC-3 cells was inhibited by oridonin solution in a concentration- and time-dependent manner; and the difference from the control group was of statistical significance (P<0.01), the IC50 value of oridonin was 10 μmol/L. Secondly, the migration and invasion of PC-3 cells were inhibited by oridonin in a concentration-dependent manner; and the difference from the control group was of statistical significance (P<0.01). Finally, the results of qRT-PCR showed that oridonin solution promoted the expression of miR-204-5p in PC-3 cells and blocked the expression of the downstream genes in the NF-κB signaling pathway; the results of Western blotting indicated that both the overexpression of miR-204-5p and oridonin could inhibit the expression of the downstream genes in NF-κB signaling pathway in PC-3 cells. Conclusions Oridonin can inhibit the proliferation, migration and invasion of PC-3 cells. Its mechanism of action may be associated with its up-regulating miR-204-5p and blocking the NF-κB signaling pathway.