摘要
目的研究吡咯烷二硫代氨基甲酸盐(PDTC)对喉癌Hep-2细胞增殖和凋亡的影响。方法体外培养Hep-2细胞,细胞分为3个组:对照组、实验分为50、100μmol/L组。采用四甲基偶氮唑蓝(MTT)实验检测不同浓度PDTC对Hep-2细胞生长的抑制作用;流式细胞术检测PDTC对Hep-2细胞凋亡的影响;实时定量聚合酶链反应(RT-PCR)检测PDTC对Hep-2细胞Bcl-xL和XIAP的mRNA表达的影响;蛋白印迹(Western-blot法)检测p-IκB、Bcl-xL及XIAP的蛋白表达及活化。结果 PDTC对50μmol/L组和100μmol/L组的Hep-2细胞生长抑制与对照组比较,差异有统计学意义,PDTC的浓度越大,抑制效果越明显(P<0.05)。Hep-2细胞经PDTC处理24h后,50μmol/L组和100μmol/L组细胞凋亡率分别为(25.1±2.18)%和(35.4±2.62)%,与对照组(3.12±1.07)%比较,差异有统计学意义。PDTC能导致Hep-2细胞生长抑制和凋亡增加且呈量效关系。结论 PDTC能够通过阻断Hep-2细胞NF-κB的活化,降低与抗凋亡相关基因Bcl-xL及XIAP的表达,抑制Hep-2细胞增殖并促进凋亡。
Objective To observe the effects of ammonium pyrrolidinedithiocarbamat（PDTC） to apoptosis and proliferation of Hep-2 cells.Methods Hep-2 cells were cultured in vitro.The control group and PDTC treatment groups were established according to the experimental requirements.The methy1 thiazoly1 tetrazolium（MTT） assay was used to measure the growth inhibition rate of Hep-2 cells.The flow cytometry assay was used to determine the apoptosis in Hep-2 cells.Bcl-xL and XIAP mRNA levels were measured by real time PCR in untreated Hep-2 cells and cells treated for varying amounts of PDTC.Western-blot analysis was used to determine the protein levels of p-IκB,Bcl-xL and XIAP in Hep-2 cells.Results The 50 μmol/L and 100 μmol/L PDTC groups inhibited the growth of Hep-2 cells,difference compared with the control group showing statistical significance.The greater the PDTC concentration was,the more significant the inhibition effect was（P0.05）.After 24 h treating by PDTC,the apoptosis rate of Hep-2 cells in the 50 μmol/L and 100 μmol/L PDTC groups were（25.1±2.18）% and（35.4±2.62）% respectively,difference compared with the control group[（3.12±1.07）%] showing statistical significance.PDTC could lead to the growth inhibition of Hep-2 cells and the increase of apoptosis with the dose-effect relationship.Conclusion PDTC inhibits the proliferation of Hep-2 cells and promotes their apoptosis by blocking the NF-κB activation of Hep-2 cells and reducing the expression of Bcl-xL and XIAP.
出处
《重庆医学》
CAS
CSCD
北大核心
2013年第9期1002-1004,共3页
Chongqing medicine
