S100A4反義寡核苷酸抑制神經(jīng)母細(xì)胞瘤細(xì)胞侵襲的實(shí)驗(yàn)研究.pdf

1、方法:該研究以兩種高表達(dá)S100A4基因的神經(jīng)母細(xì)胞瘤細(xì)胞系LA-N-6和SK-N-SH為體外模型,以人S100A4 mRNA序列5'翻譯起始位點(diǎn)第70~89位堿基為反義作用靶點(diǎn),人工合成20聚反義寡核苷酸(antisense oligodeoxynucleotide,asODN),并以其正義鏈和錯(cuò)義鏈為對(duì)照,通過脂質(zhì)體包裹的方法進(jìn)行細(xì)胞轉(zhuǎn)染.采用半定量逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(reverse transcription-polymeras

2、e chain reaction,RT-PCR)方法在mRNA水平檢測(cè)asODN對(duì)兩種神經(jīng)母細(xì)胞瘤細(xì)胞S100A4表達(dá)的抑制作用和對(duì)MMP-2表達(dá)的影響,以比色法檢測(cè)asODN對(duì)兩種神經(jīng)母細(xì)胞瘤細(xì)胞的細(xì)胞活力的影響,以Transwell濾膜小室法檢測(cè)神經(jīng)母細(xì)胞瘤細(xì)胞體外侵襲和運(yùn)動(dòng)能力.結(jié)論:脂質(zhì)體介導(dǎo)的S100A4反義寡核苷酸能夠抑制神經(jīng)母細(xì)胞瘤LA-N-6和SK-N-SH細(xì)胞中S100A4 mRNA表達(dá),抑制LA-N-6細(xì)胞體外侵襲能