本文選題：沉默信息調(diào)節(jié)因子樣蛋白 + RNA干擾��； 參考：《軍事醫(yī)學(xué)》2017年02期

【摘要】：目的構(gòu)建沉默信息調(diào)節(jié)因子(silent information regulaor,Sir)樣蛋白1(Sirtuin 1,SIRT1)的特異性短發(fā)夾RNA(shRNA)慢病毒載體,獲得敲低SIRT1的肝癌細(xì)胞系,以探討其對肝癌細(xì)胞的增殖和耐藥敏感性的作用。方法設(shè)計(jì)針對SIRT1靶點(diǎn)特異性的干涉序列,連接到經(jīng)HpaⅠ和XhoⅠ雙酶切的pSicoR-GFP載體,慢病毒包裝293T產(chǎn)生病毒,感染肝癌細(xì)胞,建立肝癌細(xì)胞SIRT1低表達(dá)的穩(wěn)定株。利用實(shí)時定量PCR檢測SIRT1的干涉效果;通過平板克隆形成實(shí)驗(yàn)、CCK8細(xì)胞增殖實(shí)驗(yàn)檢測SIRT1被干涉后對肝癌細(xì)胞增殖能力的影響;通過細(xì)胞耐藥敏感性實(shí)驗(yàn)及實(shí)時定量PCR檢測耐藥基因的表達(dá),考察SIRT1干涉對肝癌細(xì)胞耐藥敏感性的影響。結(jié)果實(shí)時定量PCR實(shí)驗(yàn)證明該慢病毒干涉載體能顯著抑制肝癌細(xì)胞中SIRT1的表達(dá),SIRT1干涉可抑制肝癌細(xì)胞的增殖能力,下調(diào)耐藥基因的表達(dá),增強(qiáng)肝癌細(xì)胞的藥物敏感性。結(jié)論 SIRT1抑制肝癌細(xì)胞耐藥性。
[Abstract]:Objective to construct the lentivirus vector of silent information regulatory sir-1 (1(Sirtuin 1)-like protein of silencing information regulator, and obtain the cell line of hepatocellular carcinoma with low SIRT1 knockout, in order to investigate its effect on the proliferation and drug resistance of hepatoma cells. Methods the interference sequence specific to SIRT1 target was designed and ligated to pSicoR-GFP vector digested by Hpa 鈪，

本文編號：1862304