穿膜素TAT介导的KDR-siRNA慢病毒载体的构建及其靶向肺癌A549细胞的抗肿瘤作用

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。%In order to construct TAT and KDR siRNA lentivirus vector and observe its in vitro target-oriented anti-tumor effect against lung cancer A549 cell lines, TAT-KDR siRNA lentivirus vector was constructed adopting recombi-nant technology, and transfected into A549 human lung cancer cell lines. The gene level variation of KDR were detec-ted by real-time fluorescent quantitative PCR and Western blot;The apoptosis and cell proliferation and clone forma-tion 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 tar-get-oriented siRNA lentivirus vector possessed significant tumour target-orientation and anti-tumour activity.