目的:构建SH3结构域结合蛋白4(SH3-domain binding protein 4,SH3BP4)重组腺病毒载体及shSH3BP4慢病毒载体,初步探索SH3BP4对肝癌细胞迁移增殖能力的影响。方法:PCR扩增SH3BP4 cDNA序列并构建腺病毒重组质粒pAdEasySH3BP4,HEK293细胞包装重组腺病毒AdSH3BP4;构建SH3BP4基因短发夹型RNA干扰慢病毒载体,转染至HEK293T细胞,包装重组慢病毒shSH3BP4。采用Western blot检测SH3BP4在人正常肝脏细胞及肝癌细胞的内源性表达,将SH3BP4过表达实验部分设为3个组:Mock组、AdGFP组和AdSH3BP4组,Mock组为空白对照组,AdGFP组为感染腺病毒AdGFP的对照组,AdSH3BP4组为感染腺病毒AdSH3BP4的实验组;将SH3BP4敲低实验部分设为2个组:sh Control组和shSH3BP4组,sh Control组为感染慢病毒sh PLL3.7的对照组,shSH3BP4组为感染慢病毒shSH3BP4的实验组。通过Transwell和划痕实验检测SH3BP4过表达和敲低对肝癌细胞迁移能力的影响...
【英文摘要】
Objective:To investigate the effect of SH3BP4 on the migration and proliferation of hepatocarcinoma cells by the construction of SH3BP4 recombinant adenovirus vector and lentiviral vector. Methods :The SH3BP4 cDNA sequence was amplified and then the recombinant Adplasmid pAdTrack-SH3BP4 was constructed. The recombinant adenovirus Ad-SH3BP4 was packaged and amplified by using HEK293 cells. The short hairpin RNA recombinant lentiviral vector of SH3BP4 was constructed and transfected into HEK293 T cells,then t...