白藜芦醇对喉癌Hep-2细胞顺铂敏感性的增强作用及其机制

doi:10.13481/j.1671-587x.20150214

摘要/Abstract

摘要：

目的:探讨白藜芦醇提高喉癌Hep-2细胞株对顺铂(DDP)敏感性的作用,并阐明其作用机制。方法:Hep-2喉癌细胞随机分为空白对照组和DDP组(终浓度分别为3、6、12和24 μmol·L^-1)培养6、12、24及48 h。CCK-8法检测各组细胞生存率;选出DDP的最佳作用浓度6 μmol·L^-1和作用时间24 h。Hep-2喉癌细胞株经培养后随机分为对照组、DDP组、白藜芦醇组、sirtinol组、白藜芦醇+sirtinol组、白藜芦醇+DDP组、sirtinol+DDP组和白藜芦醇+sirtinol+DDP组。CCK-8法检测各组细胞生存率;采用免疫荧光法观察各组细胞中Sirt1蛋白的表达强度;Annexin Ⅴ-FITC Kit法检测各组细胞凋亡率。结果:CCK-8检测,与对照组比较,DDP组和白藜芦醇组细胞生存率明显下降(P<0.01);与DDP组比较,白藜芦醇+DDP组细胞生存率明显下降(P<0.01);与白藜芦醇+sirtinol+DDP组比较,白藜芦醇+DDP组细胞生存率明显下降(P<0.05)。细胞免疫荧光法检测,与DDP组比较,白藜芦醇+DDP组细胞中Sirt1 蛋白的平均免疫荧光强度明显升高(P<0.01);与白藜芦醇+sirtinol+DDP组比较,白藜芦醇+DDP组细胞中Sirt1 蛋白的平均免疫荧光强度明显升高(P<0.01)。Annexin Ⅴ-FITC Kit检测,与DDP组比较,白藜芦醇+DDP组细胞凋亡率均明显升高(P<0.01);与白藜芦醇+sirtinol+DDP组比较,白藜芦醇+DDP组细胞凋亡率明显升高(P<0.01)。结论:白藜芦醇提高Hep-2喉癌细胞株对DDP的敏感性,其机制可能与提高喉癌细胞中Sirt1蛋白表达有关联。

关键词: 白藜芦醇, 沉默信息调节因子 1, 喉肿瘤, Hep-2细胞, 顺铂, sirtinol

Abstract:

Objective To investigate the enhancement effect of resveratrol on the sensitivity of laryngeal carcinoma Hep-2 cells to cisplatin(DDP) and its mechanism.Methods The Hep-2 cells were randomly divided into blank control group and different doses(3,6,12, and 24 μmol·L^-1)of DDP groups and the cells were treated for 6,12,24,and 48 h.The suitable concentration of DDP(6 μmol·L^-1) and culture time(24 h) were chosen.The Hep-2 cells treated under the suitable condition were randomly divided into control group,DDP group,resveratrol group,sirtinol group,resveratrol+sirtinol group,resveratrol+DDP group,sirtinol+DDP group and resveratrol+sirtinol+DDP group. The survival rates of the Hep-2 cells in various groups were detected by CCK-8 assay;the expression levels of Sirt1 protein in the Hep-2 cells in various groups were determined with immunofluorescence method;the apoptotic rates of the Hep-2 cells in various groups were detected by Annexin Ⅴ-FITC Kit.Results The CCK-8 assay results showed that the survival rates of the Hep-2 cells in resveratrol group and DDP group were decreased significantly compared with control group(P<0.01);compared with DDP group,the survival rates of the Hep-2 cells in resveratrol+DDP group were decreased(P<0.01);compared with resveratrol+DDP+sirtinol group,the survival rates of the Hep-2 cells in resveratrol+DDP group were decreased(P<0.05).The immunofluorescence results showed that the mean fluorescence intensity of Sirt1 protein of the cells in resveratrol+DDP group were increased significantly compared with DDP group(P<0.01);compared with resveratrol+sirtinol+DDP group,the mean fluorescence intensity of Sirt1 protein of the cells in resveratrol+DDP group was increased significantly (P<0.01).The Annexin Ⅴ-FITC Kit results showed the apoptotic rate of the cells in resveratrol+DDP group was higher than those in DDP group and resveratrol+sirtinol+DDP group(P<0.01).Conclusion Resveratrol can improve the sensitivity of Hep-2 cells to cisplatin,and its mechanism may be related to increasing the expression of Sirt1 protein.

Key words: resveratrol, silent information regulator 1, laryngeal neoplasms, Hep-2 cells, cisplatin, sirtinol

中图分类号:

R739.65

战仕花, 王苹, 尹万忠, 祝威. 白藜芦醇对喉癌Hep-2细胞顺铂敏感性的增强作用及其机制[J]. 吉林大学学报(医学版), 2015, 41(02): 282-286.

ZHAN Shihua, WANG Ping, YIN Wanzhong, ZHU Wei. Enhancement effect of resveratrol on sensitivity of laryngeal carcinoma Hep-2 cells to cisplatin and its mechanism[J]. Journal of Jilin University Medicine Edition, 2015, 41(02): 282-286.

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