周俐达,崔颖,董闯
ZHOU Lida,CUI Ying,DONG Chuang

• 1:锦州医科大学研究生院
• 2:锦州医科大学附属第一医院

摘要(Abstract)：

目的利用姜黄素及其联合白藜芦醇在体外作用于喉癌细胞Hep-2,观察其对人喉癌细胞的影响,并探讨可能的抗肿瘤作用机制。方法采用MTT法检测各组药物对喉癌细胞增殖的抑制作用;Hoechst 33258染色法观察各组药物对喉癌细胞核的形态影响;流式细胞术检测细胞凋亡率;RT-PCR检测p65和caspase-3的基因表达情况;Western blot法检测各组药物对喉癌细胞p65蛋白和caspase-3蛋白表达情况的影响。结果 MTT结果显示姜黄素联合白藜芦醇组抑制喉癌细胞增殖的作用要明显高于单独姜黄素组和单独白藜芦醇组(P<0.05);Hoechst 33258染色法的结果显示,经各组药物处理后的细胞核形态与对照组相比呈凋亡特征性改变;流式细胞术分析结果显示姜黄素联合白藜芦醇组的细胞凋亡率明显高于单独姜黄素组和单独白藜芦醇组(P<0.05);RT-PCR结果显示姜黄素联合白藜芦醇组的p65基因表达量与二者单独使用时相比明显降低(P<0.05),而姜黄素联合白藜芦醇组的caspase-3基因表达量与二者单独使用时相比明显升高(P<0.05);Western blot结果显示与对照组比较,单独姜黄素组和单独白藜芦醇组的p65蛋白表达明显下调,caspase-3蛋白表达则明显上调,而姜黄素联合白藜芦醇组对p65和caspase-3表达调控的影响与单独姜黄素组和单独白藜芦醇组相比差异有统计学意义(P<0.05)。结论姜黄素联合白藜芦醇在诱导人喉癌细胞Hep-2的凋亡时具有协同效应,其作用机制可能是通过阻断转录因子NF-κB(p65)信号通路的活化,上调caspase-3蛋白的表达来实现的。
Objective To observe the effect of curcumin and its combination w ith resveratrol on the proliferation and apoptosis of laryngeal cancer cells in vitro,and to investigate the possible mechanism of anti-tumor action on Hep-2. Methods Cell grow th inhibition rate w as detected by MTT method; The morphological changes of the nucleus w ere observed by Hoechst 33258 staining method; Apoptosis rate w as detected by flow cytometry; The expression of p65 and caspase-3 gene w as detected by RT-PCR; Western blot method w as uesd to detect the effect of drugs on the expression of p65 protein and caspase-3 protein in laryngeal carcinoma cells. Results The result of MTT show ed that the effect of curcumin combined w ith resveratrol on the proliferation of laryngeal cancer cells w as significantly higher than that of curcumin and resveratrol used alone( P < 0. 05); The results of Hoechst 33258 staining show ed that the morphology of nuclei in each treatment group show ed a characteristic change of apoptosis compared w ith the control group;The results of flow cytometry analysis show ed that the apoptosis rate of curcumin combined w ith resveratrol group w as significantly higher than that of curcumin and resveratrol alone group( P < 0. 05); RT-PCR results show ed that the expression of p65 gene in curcumin combined w ith resveratrol group w as significantly low er than that in the control group( P < 0. 05),and the expression of caspase-3 gene in curcumin combined w ith resveratrol group w as significantly higher than in the control group( P < 0. 05); Western blot analysis show ed that expression of p65 protein w as dow n-regulated and the expression of caspase-3 protein w as significantly up-regulated in curcumin group alone and resveratrol alone group compared w ith the control group. w hile the expression of p65 and caspase-3 in curcumin combined w ith resveratrol group had significant difference compared w ith single curcumin and resveratrol alone group( P < 0. 05). Conclusions Curcumin and resveratrol has synergistic effect on inducing apoptosis of human laryngeal carcinoma cell Hep-2,and the mechanism is probably by blocking transcription factor kappa B NF-( p65) signal pathw ay activation and upregulating expression of caspase-3 protein.

关键词(KeyWords)： 姜黄素;白藜芦醇;人喉癌细胞Hep-2;细胞凋亡;NF-κB信号通路
curcumin;resveratrol;human laryngeal carcinoma cell Hep-2;apoptosis;NF-κB signaling pathway

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 周俐达,崔颖,董闯
ZHOU Lida,CUI Ying,DONG Chuang

DOI: 10.14066/j.cnki.cn21-1349/r.2018.01.012

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