干扰Netrin-1对白血病K562细胞体内外增殖迁移的影响

Effects of Netrin-1 Interference on Proliferation and Migration of Leukemia K562 Cells in vitro and in vivo

目的探究干扰Netrin-1对白血病K562细胞体内外增殖迁移的影响。方法将shRNA Netrin-1载体和空质粒载体转染至K562细胞,细胞随机分为3组:Control组、shRNA-NC组和shRNA2-Netrin-1组。克隆形成实验检测克隆形成率;Western印迹检测Netrin-1、Ki67、Survivin蛋白表达;Transwell检测侵袭细胞数;划痕实验检测划痕愈合率;建立白血病K562细胞裸鼠移植瘤模型,检测30 d肿瘤体积和肿瘤重量变化;免疫组化检测Ki67、VEGF阳性细胞数。结果体外实验中,与Control组比较,shRNA2-Netrin-1组K562细胞克隆形成率、Ki67和Survivin蛋白表达、侵袭细胞数及划痕愈合率均降低(P<0.05);体内实验中,与Control组比较,shRNA2-Netrin-1组裸鼠肿瘤体积、肿瘤重量、Ki67阳性细胞数和VEGF阳性细胞数均降低(P<0.05)。结论敲低Netrin-1表达可抑制白血病K562细胞体内外增殖及迁移。

Objective To investigate the effect of Netrin-1 interference on the proliferation and migration of leukemia K562 cells.Methods K562 cells were transfected with shRNA Netrin-1 vector and empty plasmid vector,and the cells were randomly divided into 3 groups:control group,shRNA-NC group and shRNA2-Netrin-1 group.Clone formation assay was used to detect the clone formation rate,and Western blot to detect the expression of Netrin-1,Ki67 and Survivin proteins.Invasive cells were detected by Transwell.The scratch healing rate was measured by scratch test.A nude mouse xenograft model of leukemia K562 cells was established,and the changes of tumor volume and Tumor weight and tumor weight at 30 days were detected.Ki67 and VEGF positive cells were detected by immunohistochemistry.Results Compared with the control group,the cloneformation rate,Ki67 and Survivin protein expression,invasive cell number and scratch healing rate of K562 cells in shRNA2-Netrin-1 group were significantly decreased in vitro(P<0.05).In vivo,compared with the control group,the tumor volume,tumor weight,the number of Ki67 positive cells and the number of VEGF positive cells were significantly decreased in the shRNA2-Netrin-1 group(P<0.05).Conclusion Knockdown of Netrin-1 expression can inhibit the proliferation and migration of leukemia K562 cellsin vitroandin vivo.