摘要
背景与目的:研究表明组蛋白去乙酰化酶(histonedeacetylase,HDAC)在多种肿瘤细胞中高表达,组蛋白去乙酰化酶1(HDAC1)抑制剂制滴菌素A(trichostatinA,TSA)能抑制肿瘤细胞的生长。而缺氧是恶性肿瘤的重要特征,本实验拟研究HDAC1在低氧肺腺癌细胞株A549中的表达及TSA对细胞增殖、凋亡的影响。方法:实验设常氧组(对照组)及低氧组(实验组),实验组设A～F组,即低氧6h组(A组),低氧6h加TSA组(B组),低氧12h组(C组),低氧24h组(D组),低氧24h加TSA组(E组),低氧48h组(F组)。用蛋白印迹法(Westernblot)检测A549细胞在不同时间低氧条件下HDAC1的表达,用四甲基偶氮唑蓝(MTT)比色、免疫组化和流式细胞仪、原位凋亡(Tunel)法检测A549细胞在不同时间低氧条件下的增殖和凋亡率以及TSA对其的影响,用RT-PCR检测A549细胞在不同时间低氧条件下HDAC1mRNA的表达及TSA对其的作用。结果:对照组HDAC1蛋白质表达的A值为138±11,实验A、C、D、F组A值分别为78±4.0、86±5.0、124±3.0、120±9.0。对照组HDAC1mRNA的A值与β-actinmRNAA值的比值0.68±0.03,A、B、D、E组HDAC1mRNA的A值与β-actinmRNAA值的比值分别为0.46±0.03、0.45±0.02、0.70±0.03、0.33±0.02。对照组PCNA的A值0.13±0.03、A、B、D、E组PCNA的A值分别为0.10±0.02。
BACKGROUND &OBJECTIVE:Histone deacetylase(HDAC)shows a hig h expression in many cance r cells and the inhibitor of HDAC1,trichostatin A(TSA),can inhibit the g rowth of cancer cel ls.Hypoxia is a common feature of malig nant tumors.This pa per was desig ned to investig ate the expression of HDAC1of A549cell stra ins in hypoxia condition and the effe ct of TSA on their proliferation and apopt osis.METHODS :The authors desig ned 1normoxia g roup(control g roup)and 5hypoxia g roups (test g roups ):hypoxia 6h g roup(A),TSA +hypoxia 6h(B),hypoxia 12h g roup(C),hypoxia 24h g roup(D),TSA +hypoxia 24h(E),hypoxia 48h g roup(F).The expression of HDAC1in A549cells was examined usin g Western blot analysis.Proliferat ion,the apoptotic rates of A549cells and the effect of TSA on them were determi ned using MTT method,immunohistochemi stry,TUNEL method,and flow cytomet ry.The expression of mRNA of HDAC1and th e effect of TSA on it were determined using reverse transcription-polym erase chain reaction(RT-PCR).RESULTS :The A values expressed by HDAC1in A549cel l strains were 138±11in the control g roup,78±4,86±5,124±3,and 120±9i n test g roups A,C,D,and F,respectively.The A values of HDAC1mRNA versus the A values of β-Atin mRNA were 0.68±0.03in the control g roup,0.46±0.03,0.45±0.02,0.70±0.03,and 0.33±0.02in test g roups A,C,D,a nd F,respectively.The A values of the expression of PCNA in A549cell st rains were 0.13±0.03in the control g roup,0.10±0.02,0.11±0.02,0.16±0.02,and 0.11±0.03in test g roups A,B,D,and E,respectively.The A values of MTT in A549cell strains wer e0.50±0.06in the control g roup,0.41±0.04,0.45±0.03,0.59±0.02,and0.45±0.03in test g roups A,B,D,and E ,respectively.The A values of positive cells of apoptosis in A549c ell strains were 0.16±0.04in the con trol g roup,0.18±0.02,0.18±0.05,0.20±0.05,and 0.23±0.05in test g roups A,B,D,and E,respectively.The apop totic rates in A549cells were 1.11%i n the control g roup,18.91%,14.30%,36.99%,and 51.92%in test g r oups A,B,D,and E,respecti
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第9期922-926,共5页
Chinese Journal of Cancer
