Construction Of Recombinant Adenovirus Vector Expressing Human SiALK2and Effect Of SiALK2in Vitro On Proliferation，Migration And Invasion Abilities Of Human Breast Cancer MDA-MB-231Cells

Objective To construct recombinant adenovirus vector expressinghuman siALK2（hsiALK2）; To investigate the effect of ALK2onproliferation，migration and invasion abilities of human breast cancerMDA-MB-231cells in vitro．Lay a foundation for further study on ALK2．Methods The recombinant adenovirus plasmid pAdsiALK2wasdigested with Pac Ⅰ and transfected to HEK293packaging cells. Therecombinant Adenovirus was amplified and identificated by RT-PCR；MDA-MB-231cells were infected with adenovirus siALK2and RFPrespectively. The proliferation, migration and invasion ofMDA-MB-231/siALK2cells were estimated by MTT assay,colony-forming assay,wounding healing assay and transwell assay.Results Both recombinant shuttle plasmid Pses-Hus-siALK2andrecombinant adenovirus vector pAdsiALK2were constructedcorrectly． Recombinant adenovirus AdsiALK2was successfully packaged in HEK293cells. MDA-MB-231cells which were infected with theAdsiALK2and AdRFP after36h have the same amount of fluorescenceexpression. ALK2expression was remarkably decreased inMDA-MB-231/siALK2cells． The proliferation activity，colony formationrate，wound healing rate and count of cells crossing the matrix barrier weresignificantly reduced in MDA-MB-231/siALK2group than those inMDA-MB-231/RFP group（P＜0.05），while no significant difference wasobserved between MDA-MB-231/RFP and blank control groups（P＞0.05）．Conclusions The recombinant adenovirus vector expressinghsiALK2was successfully constructed; down-regulating ALK2expression can inhibite the proliferation， migration and invasion ofMDA-MB-231cells in vitro．...