曲古抑菌素A对鼻咽癌CNE3细胞增殖抑制作用

目的 探讨曲古抑菌素A(TSA)对鼻咽癌CNE3细胞周期阻滞与凋亡影响及作用机制。方法 以不同浓度TSA作用鼻咽癌CNE3细胞株,采用甲基噻唑基四唑(MTT)法检测细胞生长抑制率;流式细胞术测定TSA作用前后鼻咽癌细胞调亡情况及细胞周期变化;逆转录-聚合酶链反应(RT-PCR)分析鼻咽癌细胞中细胞周期素依赖性蛋白激酶相互作用蛋白1(p21)mRNA表达的变化。结果 100～500 nmol/L TSA对CNE3细胞生长有明显抑制作用,呈剂量依赖性,抑制率为17.74%～44.33%;400,500 nmol/L TSA可在DNA合成期(S)、DNA合成后期/有丝分裂期(G₂/M)诱导CNE3细胞周期阻滞,并诱导细胞凋亡,24 h凋亡率为(14.67±0.21)%,(18.73±1.80)%,48 h凋亡率分别为(16.3±0.96)%,(39.17±1.27)%,与对照组(9.16±1.05)%比较差异均有统计学意义(P<0.05);TSA可诱导CNE3细胞内p21基因表达。结论 TSA可明显抑制CNE3细胞增殖,诱导细胞凋亡,其作用机制与p21基因异常表达有关。

Effect of TSA on nasopharyngeal carcinoma CNE3 cells and its mechanism

Objective To investig ate the effect of trichostatin A(TSA)on cell proliferation and cell cycle progression of nasopharyngeal carcinoma line CNE3,and to explore its potential mechanism sinvo lved.Methods CNE3 cells in logarithmic growth phase were incubated with TSA at different concen trations for 24 or 48 hours,respectively.The proliferation of CNE3 cells was measured with methyl thiazolyl tetrazolium(MTT)assay.The effect of TSA on cell cycle and apoptosis was detected by flowcy tometry(FCM).The expression of cyclin dependent kinase interacting prote in 1(CIP1,p21)was determined with RT-PCR.Results A fter 100-500nmol/L TSA admin istration,CNE3 proliferation inhibition ratioin creased significantly from 17.74% to 44.33% in concentration dependent manner(P<0.05).CNE3 cells showed DNA synthesis phase,second gap/mitosis phase/(S,G₂/Mphase)blockage 24.48 hours after the treamtent of TSA,and the apoptosis ratio increased.The apoptosis rate at different concentrations of TSA(400 and 500nM)were 14.67±0.21,18.73±1.80 in 24hr,and 16.3±0.96,39.17±1.27 in 48hr(P<0.05).The expression level of p21 was increased in dose dependent manner after TSA trea mtent.Conclusion TSA inhibits the proliferation of CNE3 cells and blocks the CNE3 cell cycle progression in S and G₂/Mphase.Itsmechanism may be related with the abnormal expression of p21.