β-拉帕醌抑制下咽癌Fadu细胞增殖和迁移机制的研究

Research on the mechanism ofβ-lapachone inhibiting proliferation and migration in hypop-haryngeal cancer Fadu cells

目的探讨β-拉帕醌(β-lapachone)体外抑制下咽癌Fadu细胞增殖、迁移及诱导凋亡的作用及机制。方法不同浓度的β-拉帕醌(0,1,2,3,4,5,6μmol/L)作用Fadu细胞48 h,0μmol/L组为空白对照组,应用噻唑蓝(MTT)实验及平板克隆实验检测不同浓度的β-拉帕醌对Fadu细胞增殖的影响。Transwell实验检测不同浓度的β-拉帕醌(0,2,4,6μmol/L)对Fadu细胞迁移能力的影响。流式细胞术(AnnexinⅤ-FITC/PI双染法)检测不同浓度的β-拉帕醌(0,2,4,6μmol/L)处理Fadu细胞后凋亡的变化。Western blot法检测不同浓度的β-拉帕醌(0,2,4,6μmol/L)处理Fadu细胞后其上皮-间质转化(epithelial-mesenchymal transition,EMT)及凋亡分子相关蛋白的变化。结果MTT结果显示,β-拉帕醌可显著降低Fadu细胞的存活率,对Fadu细胞具有浓度依赖性的增殖抑制作用(P<0.05),作用48 h的IC 50值为3.547μmol/L。克隆形成结果显示,β-拉帕醌可浓度依赖性地抑制Fadu细胞的增殖能力(P<0.05)。Transwell结果显示,经β-拉帕醌处理后,Fadu细胞的迁移能力明显下降,且呈浓度依赖性(P<0.05)。流式细胞术分析结果表明,β-拉帕醌可显著促进Fadu细胞的凋亡,且呈浓度依赖性(P<0.05)。Western b1ot结果显示,β-拉帕醌可下调vimentin、snail和Bcl-2蛋白的表达,并上调E-cadherin、Bax蛋白的表达(P<0.05),且呈浓度依赖性。结论β-拉帕醌对下咽癌细胞Fadu具有明显的抑制增殖及诱导凋亡的作用,并可通过EMT途径抑制下咽癌细胞的迁移能力。

Objective To investigate the effects ofβ-lapachone on the proliferation and migration and the apoptosis of hypopharyngeal cancer cells(Fadu)in vitro.Methods Fadu cells were treated with different concentrations ofβ-lapachone(0,1,2,3,4,5,6μmol/L)for 48 h,and then the effects ofβ-lapachone on the proliferation of Fadu cells were detected by MTT assay and plate cloning assay.Transwell assay was used to detect the effect of different concentrations ofβ-lapachone(0,2,4,6μmol/L)on Fadu cell migration.Flow cytometry(AnnexinⅤ-FITC/PI double staining method)was used to detect the Fadu cell apoptosis after treatment with different concentrations ofβ-lapachone(0,2,4,6μmol/L).Western blot assay was used to detect the changes of epithelial-mesenchymal transition(EMT)-and apoptosis-related protein(EMT)in Fadu cells treated with different concentrations ofβ-lapachone(0,2,4,6μmol/L).Results MTT results showed thatβ-lapachone significantly reduced the survival rate of Fadu cells and had a concentration-dependent inhibitory effect on Fadu cell proliferation(P<0.05),with an IC 50 value of 3.547μmol/L at 48 h.The results of clone formation showed thatβ-lapachone inhibited the proliferation of Fadu cells in a concentration-dependent manner(P<0.05).Transwell results showed that the migration ability of Fadu cells was decreased significantly in a concentration-dependent fashion after treatment withβ-lapachone(P<0.05).The results of flow cytometry analysis showed thatβ-lapaquinone significantly promoted the apoptosis of Fadu cells in a concentration-dependent manner(P<0.05).Western blot results showed thatβ-lapachone down-regulated the expression of vimentin,snail and Bcl-2 proteins,and up-regulated the expression of E-cadherin and Bax proteins in a concentration-dependent manner(P<0.05).Conclusionβ-lapachone can significantly inhibit the proliferation and induce the apoptosis of hypopharyngeal carcinoma cells,and can inhibit the migration ability of hypopharyngeal carcinoma cells through the EMT pathway.