人siALK2重组腺病毒的构建及其对人乳腺癌细胞MDA-MB-231增殖、迁移和侵袭的影响  

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

作　　者：费嫦[1] 王林[1] 孙笑笑[1] 刘月红[1] 万绍恒[1] 王维[1] 陈莹莹[1] 王婷[1] 张彦[1] 

机构地区：[1]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2013年第8期1166-1172,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81172017)资助的课题~~

摘　　要：用前期设计并合成好的重组腺病毒质粒pAdsiALK2,经Pac I酶切后转染至HEK293细胞进行包装,制备重组腺病毒,扩增后进行病毒滴度测定,以重组腺病毒siALK2(AdsiALK2)感染MDA-MB-231细胞,以含RFP的腺病毒作为感染对照,并设空白对照组。通过RT-PCR验证MDA-MB-231细胞中ALK2表达显著下降;MTT法、平板集落形成试验、细胞划痕试验及Transwell侵袭试验证实MDA-MB-231/siALK2组相比MDA-MB-231/RFP组细胞的增殖活力、集落形成率及划痕愈合率均显著降低(P<0.05),穿膜细胞数也明显减少(P<0.05),而MDA-MB-231/RFP组与空白对照组比较,差异无统计学意义(P>0.05)。该研究表明,下调ALK2表达后可以在体外抑制乳腺癌MDA-MB-231细胞的增殖、迁移与侵袭。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 pro- liferation activity, colony formation rate, wound healing rate and count of cells crossing the matrix barrier were signifi- cantly 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 MDA- MB-231/RFP and blank control groups （P〉0.05）. These results demonstrated that down-regulating ALK2 expressioncan inhibite the proliferation, migration and invasion ofMDA-MB-231 cells in vitro.

关 键 词：siALK2 重组腺病毒 乳腺肿瘤 细胞增殖 细胞运动 肿瘤浸润 

分 类 号：R737.9[医药卫生—肿瘤]