【摘要】 为了构建TAT与KDR-siRNA慢病毒载体,观察其对肺癌细胞株A549的体外靶向抗肿瘤作用,利用重组技术构建TAT-KDR siRNA慢病毒载体并转染人肺癌细胞株A549。实时荧光定量PCR、Western blot检测KDR基因水平变化;流式细胞仪、MTT法、集落形成试验检测其对A549细胞株细胞凋亡、细胞增殖和克隆形成的影响;细胞黏附实验评价其肿瘤靶向性。其抗癌作用主要表现为可有效地抑制A549细胞KDR基因表达、细胞增殖和克隆形成,促进细胞凋亡,并具有肿瘤靶向性作用。因而认为,TAT与KDR靶向siRNA慢病毒载体具有显著的肿瘤靶向性和抗肿瘤活性。更多还原

【Abstract】 In order to construct TAT and KDR siRNA lentivirus vector and observe its in vitro target-oriented antitumor effect against lung cancer A549 cell lines,TAT-KDR siRNA lentivirus vector was constructed adopting recombinant technology,and transfected into A549 human lung cancer cell lines. The gene level variation of KDR were detected by real-time fluorescent quantitative PCR and Western blot; The apoptosis and cell proliferation and clone formation of A549 cell lines were determined by MTT method or Flow cytometry or colony forming test; The evaluation of tumor target-orientation was detected by cell adhesion experiments. The anti-tumour effects were mainly reflected the effective inhibition of KDR gene expressions of A549 cell,cell proliferation,and clone formation,the promotion of cell apoptosis,and possessed the effect of tumour target-orientation. Therefore,it is believed that TAT and KDR target-oriented siRNA lentivirus vector possessed significant tumour target-orientation and anti-tumour activity.更多还原