本文選題：超聲靶向微泡破滅 + 缺氧誘導(dǎo)因子1α　； 參考：《華中科技大學(xué)》2014年博士論文

【摘要】：背景缺氧誘導(dǎo)因子la(Hypoxia-inducible factor-1alpha, HIF-la)基因的激活和過表達(dá)是經(jīng)導(dǎo)管肝動(dòng)脈栓塞術(shù)(transcatheter arterial embolization, TAE)后肝癌復(fù)發(fā)和轉(zhuǎn)移的重要因素,提高TAE的治療效果則必須抑制HIF-1α的高表達(dá)。 目的超聲轉(zhuǎn)染HIF-1α shRNA至肝癌細(xì)胞內(nèi),抑制HIF-1α的表達(dá),提高TAE的治療效果。 方法1、體外缺氧培養(yǎng)大鼠肝癌細(xì)胞walker256,將靶向針對(duì)HIF-1α的短發(fā)夾狀RNA(small hairpin RNA, shRNA)即HIF-1α shRNA表達(dá)質(zhì)粒與超聲微泡Sonovue混合后,利用超聲靶向微泡破壞(ultrasound targeted microbubbles destruction, UTMD)技術(shù)將質(zhì)粒轉(zhuǎn)染至細(xì)胞內(nèi),觀察HIF-1α的沉默效果。2、評(píng)估UTMD體內(nèi)轉(zhuǎn)染時(shí)不同條件對(duì)大鼠的損傷效應(yīng)；評(píng)估安全的轉(zhuǎn)染條件下介導(dǎo)eGFP肝內(nèi)轉(zhuǎn)染時(shí)的轉(zhuǎn)染效果,有助于選擇安全、有效的轉(zhuǎn)染條件。3、構(gòu)建wistar大鼠肝癌模型。經(jīng)大鼠尾靜脈注入shRNA表達(dá)質(zhì)粒與SonoVue的混合液,同時(shí)在大鼠肝癌部位用合適的超聲條件擊碎微泡,將shRNA質(zhì)粒釋放入細(xì)胞,觀察沉默HIF-1α基因表達(dá)的效果和腫瘤大小的變化；超聲輻照完畢后行TAE。研究中設(shè)置不同的實(shí)驗(yàn)組：對(duì)照(control)組、UTMD組、TAE組、UTMD+TAE組,對(duì)比分析各組對(duì)肝癌的治療效果。 結(jié)果1、體外實(shí)驗(yàn)證實(shí)HIF-1α shRNA能夠有效抑制缺氧條件下walker256肝癌細(xì)胞內(nèi)HIF-1α的表達(dá)；2、UTMD介導(dǎo)基因肝內(nèi)轉(zhuǎn)染時(shí),SonoVue劑量為4ul/g條件下,輻照條件為1MHz,20%DC,輻照6min時(shí),超聲強(qiáng)度≤1.5w/cm2對(duì)大鼠無明顯損傷,1.5w/cm2輻照6min可以獲得較好的轉(zhuǎn)染效果。3、體內(nèi)實(shí)驗(yàn)結(jié)果顯示：control組腫瘤持續(xù)增大,TAE組肝癌前期腫瘤生長(zhǎng)明顯受抑制,后期出現(xiàn)復(fù)發(fā),UTMD組肝癌生長(zhǎng)抑制較明顯,TAE聯(lián)合UTMD組腫瘤生長(zhǎng)抑制顯著,體積明顯縮小,達(dá)到治愈肝癌的效果。 結(jié)論UTMD能有效介導(dǎo)HIF-la shRNA轉(zhuǎn)染至細(xì)胞內(nèi),沉默HIF-1α的表達(dá),聯(lián)合TAE能顯著抑制腫瘤生長(zhǎng),癌細(xì)胞壞死徹底,能夠達(dá)到治愈肝癌、防止復(fù)發(fā)的目的,為肝癌治療提供新的方法和思路。
[Abstract]:Background the activation and overexpression of Hypoxia-inducible factor-1 alpha (HIF-la-1) gene is an important factor in the recurrence and metastasis of hepatocellular carcinoma (HCC) after transcatheter arterial embolization (TAEs). To increase the therapeutic effect of Tae, we must inhibit the high expression of HIF-1 偽. Objective to transfect HIF-1 偽 shRNA into hepatoma cells and inhibit the expression of HIF-1 偽. Methods 1. Rat hepatoma cell line walker 256 was cultured under hypoxia. The short hairpin targeting HIF-1 偽, HIF-1 偽 shRNA expression plasmid, was mixed with ultrasound microbubble Sonovue. The silencing effect of HIF-1 偽 was observed by ultrasound targeted microbubble destruction ultrasound targeted microbubbles destruction, UTMD-mediated transfection into the cells, and the damage effects of different transfection conditions on the rats were evaluated by observing the silencing effect of HIF-1 偽. To evaluate the transfection effect of intrahepatic transfection of eGFP under the safe transfection condition, it is helpful to select the safe and effective transfection condition. 3. To construct the hepatoma model of wistar rats. The mixture of shRNA expression plasmid and SonoVue was injected into the tail vein of rats. At the same time, the shRNA plasmid was released into the cells with appropriate ultrasound condition. The effect of silencing HIF-1 偽 gene expression and the change of tumor size were observed. After ultrasonic irradiation, TAE was used. In the study, different experimental groups were set up: control group (control group), UTMD group (Tae group), Results 1. In vitro experiments showed that HIF-1 偽 shRNA could effectively inhibit the expression of HIF-1 偽 in walker256 hepatoma cells under hypoxia. When 6min was irradiated, ultrasound intensity 鈮，

本文編號(hào)：1999858