藏红花素对人宫颈癌Hela细胞活力、凋亡和细胞周期分布的影响

Effects of Crocin on the viability,apoptosis and cell cycle of human cervical cancer Hela cell

目的探讨藏红花素对人宫颈癌Hela细胞活力、凋亡和细胞周期分布的影响及其机制。方法体外培养并取对数生长期人宫颈癌Hela细胞,分别给予藏红花素低、中、高剂量(200,400,800 mg/L)进行干预,并设空白对照组和顺铂(0.8 mg/L)组。CCK-8法检测细胞活力,Annexin V-FITC/PI双染法检测细胞凋亡情况并计算凋亡率,流式细胞术检测细胞周期分布情况,Western blot法检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和细胞周期素B1(Cyclin B1)、周期蛋白依赖激酶1(CDK1)蛋白表达情况。结果与空白对照组比较,藏红花素中、高剂量组及顺铂组人宫颈癌Hela细胞活力均明显降低(P均<0.05),细胞凋亡率均明显增高(P均<0.05),G0/G1期细胞比例增高而G2/M期细胞比例降低(P均<0.05),Bcl-2蛋白表达量明显降低(P<0.05),Bax、Cyclin B1、CDK1蛋白表达量及Bax/Bcl-2比值均明显增高(P均<0.05),且藏红花素高剂量组上述指标变化幅度均明显高于顺铂组(P均<0.05)。结论藏红花素具有降低人宫颈癌Hela细胞活力、促进凋亡、阻滞细胞周期进程的作用,作用机制可能与调节细胞凋亡相关蛋白和细胞周期相关蛋白表达有关。

Objective It is to investigate the effects of Crocin on the viability,apoptosis and cell cycle distribution of human cervical cancer Hela cell and its mechanism.Methods The human cervical cancer Hela cells in logarithmic growth period were cultured in vitro and obtained,they were given low,medium,and high doses of Crocin(200,400,800 mg/L)for intervention,and a blank control group and cisplatin(0.8 mg/L)group were set up.The cell viability was detected by CCK-8 method;the apoptosis status was detected by Annexin V-FITC/PI double staining method and the apoptosis rate was calculated;the cell cycle distribution was detected by flow cytometry;the expression of B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 related X protein(Bax),Cyclin B1(Cyclin B1),and cyclin-dependent kinase 1(CDK1)protein were detected by Western blot.Results Compared with the blank control group,the viability of human cervical cancer Hela cells in the medium and high dose groups of Crocin and the cisplatin group were significantly reduced(all P<0.05),and the apoptosis rate was significantly increased(all P<0.05),the ratio of cells in G0/G1 phase was increased while the ratio of cells in G2/M phase was decreased(all P<0.05),the Bcl-2 protein expression was significantly reduced,and the Bax,Cyclin B1,CDK1 protein expression and the Bax/Bcl-2 ratio were all significantly increased(P<0.05),and the range of changes in the above indicators in the crocin high-dose group was significantly higher than that in the cisplatin group(all P<0.05).Conclusion Crocin has pharmacological effect that reduces the viability of human cervical cancer Hela cells,promotes apoptosis and blocks the progress of the cell cycle;which mechanism may be related to the regulation of cell apoptosis-related proteins expression and cycle-related proteins expression.