串珠素反义cDNA质粒转染对喉癌细胞Hep-2增殖能力的影响  

Suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA

作　　者：陈广理[1] 龚树生[1] 丁娟[1] 刘英鹏[1] 罗凌惠[1] 王建亭[1] 陈沛[1] 鄢开胜[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院耳鼻咽喉科

出　　处：《临床耳鼻咽喉科杂志》2006年第21期984-987,共4页Journal of Clinical Otorhinolaryngology

基　　金：第4届教育部"高校青年教师奖"资助项目

摘　　要：目的:研究串珠素反义cDNA质粒转染对喉癌细胞Hep-2增殖能力的影响。方法:利用阳离子脂质体作为载体,把重组真核表达载体串珠素反义cDNA质粒pAP转染喉癌细胞Hep-2,通过RT-PCR、Westernblot及MTT法检测转染后Hep-2细胞串珠素mRNA、蛋白质表达及细胞增殖水平,并观察转染后Hep-2细胞对碱性成纤维细胞生长因子(bFGF)的反应性。将Hep-2细胞分3组:①未转染的Hep-2细胞组(WT组);②空载体phβApr-neo1转染组(neo组);③串珠素反义cDNA质粒pAP转染组(pAP组)。结果:将质粒pAP成功地转入Hep-2细胞,获得了稳定表达串珠素反义cDNA基因的细胞系。串珠素mRNA和蛋白质水平在pAP组低表达,在WT组和neo组高表达,均差异有统计学意义(均P<0.01)。在0.1%FCS的RPMI1640培养基培养下,WT组、neo组和pAP组细胞的增殖率均降低,但pAP组细胞的增殖与WT组和neo组细胞相比明显减低;在0.1%FCS加1μg/LbFGF的RPMI1640培养液培养下,WT组和neo组细胞增殖接近正常,但pAP组细胞的增殖能力较弱。结论:串珠素反义cDNA质粒转染可以有效抑制喉癌细胞Hep-2的增殖能力。Objective： To study the suppression of proliferation of Hep-2 ceils by stable expression of antisense perlecan cDNA. Method：In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA （pAP） were transfected into Hep-2 cells by using cationic liposome （lipofectamine 2000） and divided into three groups： non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid,pAP group. Semi quantify RT PCR,western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor（bFGF）. Result：It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph βApr-neol transfectedgroup （P〈0.01）. In the presence of 1μg/I, of bFGF in low serum （0. 1% FCS）, the pAP transfected cells showed a reduced proliferation rate （MTT assay） while the wild type cells and ph β Apr-neol trans fected cells grew rapidly. Conclusion： The growth of Hep-2 cells could be inhibited significantly by perlecan antisense cDNA plasmids transfection.

关 键 词：喉肿瘤 串珠素 反义CDNA 转染 

分 类 号：R739.65[医药卫生—肿瘤]