大黄素对宫颈癌细胞体外转移活性和顺铂敏感性的影响及其机制研究

Effects of emodin on metastatic ability and sensitivity to cisplatin of cervical cancer cells in vitro and its underlying mechanisms

摘要:

目的 探究大黄素对宫颈癌细胞体外转移活性和顺铂敏感性的影响，以期为宫颈癌的治疗提供新思路。方法 将HeLa细胞分为4组：对照组、顺铂组、大黄素组、联合组。采用Transwell实验、MTT法、流式细胞术、划痕试验检测细胞侵袭、增殖、凋亡和迁移能力，Western blotting检测大黄素对PI3K/Akt信号通路的影响。结果 对照组、顺铂组、大黄素组、联合组细胞24、36和72 h的吸光度（OD）值比较，经重复测量设计的方差分析，结果 ①不同时间点OD值比较，差异有统计学意义（F =396.000，P =0.000）；②各组细胞OD值比较，差异有统计学意义（F =158.500，P =0.000），联合组OD值低于对照组、大黄素组、顺铂组（P <0.05）；③各组细胞OD值的变化趋势比较，差异有统计学意义（F =39.100，P =0.000）。与对照组比较，大黄素组和顺铂组宫颈癌细胞集落形成数、细胞侵袭数和迁移率均减少（P <0.05）；与对照组、大黄素组、顺铂组比较，联合组的宫颈癌细胞集落形成数、细胞侵袭数和迁移率均减少（P <0.05）。与对照组比较，大黄素组和顺铂组宫颈癌细胞凋亡率升高（P <0.05）；与对照组、大黄素组、顺铂组比较，联合组宫颈癌细胞凋亡率升高（P <0.05）。与对照组比较，大黄素组和顺铂组PI3K和Akt蛋白磷酸化水平均降低（P <0.05）；与对照组、大黄素组、顺铂组比较，联合组PI3K和Akt蛋白磷酸化水平均降低（P <0.05）。结论 大黄素可以调节宫颈癌细胞增殖、侵袭和迁移能力，促进宫颈癌细胞凋亡，并提高宫颈癌细胞对顺铂的敏感性，其作用可能与PI3K/Akt信号通路有关。

Abstract:

Objective To explore the effects of emodin on metastatic ability and sensitivity to cisplatin of cervical cancer cells in vitro and to provide novel insights for the treatment of cervical cancer.Methods HeLa cells were divided into control group, cisplatin group, emodin group and combination group. The cell invasion, proliferation, apoptosis and migration were detected by transwell assay, MTT assay, flow cytometry, and scratch assay. The effect of emodin on the PI3K/Akt signaling pathway was analyzed by Western blotting.Results The optical density (OD) at 24 h, 36 h and 72 h in the control group, cisplatin group, emodin group and combination group was compared via the repeated measures ANOVA, and the results revealed that the OD was different among the time points (F =396.000, P =0.000) and the groups (F = 158.500, P = 0.000), and that the OD in the combination group was lower than that in the control group, cisplatin group and emodin group (P < 0.05). Besides, The change trend of OD was also different among the groups (F = 39.100, P = 0.000). Compared with the control group, the number of colonies of cervical cancer cells, the number of invasive cells, and the rate of migration in the emodin group and the cisplatin group were lower (P < 0.05). The number of colonies of cervical cancer cells, the number of invasive cells, and the rate of migration in the combination group were lower than those in the other groups (P < 0.05). In comparison with the control group, the apoptosis rate of cervical cancer cells was higher in the emodin group and cisplatin group (P < 0.05), while the apoptosis rate of cervical cancer cells in the combination group was higher than that in the other three groups (P < 0.05). The phosphorylation levels of the PI3K and Akt proteins were lower in the emodin group and cisplatin group relative to those in the control group (P < 0.05), whereas they were lower in the combination group than in the rest groups (P < 0.05).Conclusions Emodin might regulate the growth, invasion and migration of cervical cancer cells. In addition, emodin could induce apoptosis of cervical cancer cells and increase the sensitivity of cervical cancer cells to cisplatin, which is potentially attributed to the modulation of the PI3K/Akt signaling pathway.