基于AKT信号通路探讨紫草素对宫颈癌细胞凋亡及Cyclin-D1蛋白表达的影响

Influence of Shikonin on the Expressions of Cyclin-D1 Protein and the Apoptosis of Cervical Cancer Cells Based on AKT Signaling Pathway

目的:研究紫草素激活AKT信号通路对宫颈癌细胞凋亡及Cyclin-D1蛋白表达的影响。方法:将宫颈癌海拉(Hela)细胞接种于DEME培养基中,根据分组,加入不同浓度的紫草素细胞培养液进行培养。分别测定培养24、48、72 h后空白组、低浓度组、中浓度组及高浓度组细胞凋亡率。另采用实时荧光定量PCR技术及Western Blotting法分别检测培养24 h后各组AKT、p-AKT及细胞周期蛋白D1信使核糖核酸(cyclin-Dlmessenger RNA,Cyclin-D1mRNA)蛋白表达量。结果:低浓度组、中浓度组、高浓度组细胞凋亡率均随着时间推移而升高,其中高浓度组最显著。细胞AKT、p-AKT、Cyclin-D1 mRNA表达量及蛋白量随着浓度的增高而减少,高浓度组明显低于其他各组(P<0.05),中浓度组明显低于低浓度及空白组(P<0.05),低浓度组低于空白组(P<0.05)。结论:紫草素对宫颈癌细胞具有明显抑制作用,作用机制与其抑制AKT信号通路及Cyclin-D1表达量有关。

Objective:To study the effects of shikonin activating AKT signaling pathway on the expressions of Cyclin-D1 protein and the apoptosis of cervical cancer cells.Methods:After inoculating in DEME medium,Hela cells of cervical cells were cultured by adding different concentrations of shikonin cell culture solution according to the groups.To detect cellular apoptosis rates of the blank group,low,moderate and high concentrations groups after culturing for 24,48 and 72 hours respectively.Real-time PCR and Western blotting were adopted to measure the expressions of AKT,p-AKT and Cyclin-D1mRNA proteins in different groups after culturing for 24 hours respectively.Results:The apoptosis rate of low,moderate and high concentrations groups increased as time went by,among them,high concentrations group was the most significant.The expressions of AKT,p-AKT,Cyclin-D1 mRNA and the protein reduced as the concentrations increased,high concentration group was notably lower than other groups(P<0.05),moderate concentration group was lower than low concentration group and the blank group(P<0.05),low concentration group was lower than the blank group(P<0.05).Conclusion:Shikonin owns notable inhibitory effects on cervical cancer cells,and its mechanism might be related to that it could inhibit AKT signaling pathway and the expressions of Cyclin-D1 protein.