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Construction of Recombinant Adenovirus Vector Expressing Human siALK2 and Effect of siALK2 on Proliferation， Migration and Invasion Abilities of Human Breast Cancer MDA-MB-231 Cells
Fei Chang, Wang Lin, Sun Xiaoxiao, Liu Yuehong, Wan Shaoheng, Wang Wei, Chen Yingying, Wang Ting, Zhang Yan*
Key Laboratory of LabLaboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China
Abstract: The recombinant adenovirus plasmid pAdsiALK2 was digested with Pac I and transfected to HEK293 packaging cells. The recombinant adenovirus was amplified and identificated， MDA-MB-231 cells were infected with adenovirus siALK2 and RFP respectively. RT-PCR demonstrated that ALK2 expression was remarkably decreased in MDA-MB-231/siALK2 cells. The proliferation， migration and invasion of MDA-MB-231/siALK2 cells were estimated by MTT assay， colony-forming assay， wounding healing assay and transwell assay showed that the proliferation activity， colony formation rate， wound healing rate and count of cells crossing the matrix barrier were significantly reduced in MDA-MB-231/siALK2 group than those in MDA-MB-231/RFP group (P<0.05)， ALK2 expression was remarkably decreased in MDA-MB-231/siALK2 cells. But no significant difference was observed between MDAMB-231/RFP and blank control groups (P>0.05). These results demonstrated that down-regulating ALK2 expression can inhibite the proliferation， migration and invasion of MDA-MB-231 cells in vitro.
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