PS-341对人大肠癌细胞LoVo,SW480中NF-κB、uPA表达的影响及其与侵袭力的关系

Effect of PS-341 on the Expression of NF-κB and uPA as well as Cell Invasiveness in Human LoVo,SW480 Colon Cancer Cells

目的观察人大肠癌细胞LoVo、SW480中真核细胞转录因子-κB(NF-κB)、尿激酶型血浆纤溶酶原激活因子(uPA)的表达差异及硼替佐米(bortezomib,PS-341)对其表达的影响及其与侵袭力的关系。方法实验分为4组:对照组(无血清)、血清组(10%FCS)、PS-341组[10%FCS+PS-341(4ng/mL)]、PDTC(NF-κB抑制剂)组[10%FCS+PDTC(10-2 mol/L)]。Western-blot测定人大肠癌细胞LoVo、SW480中NF-κB、uPA的蛋白表达水平;Brdu ELISA法测定细胞增殖活性;Boyden小室培养测定细胞的迁移。结果 (1)与SW480细胞比较,血清组中LoVo细胞总NF-κB、核内NF-κB和uPA表达、细胞增殖及细胞迁移率均显著增高(P<0.05);(2)与对照组比较,血清组中2种细胞总NF-κB、核内NF-κB和uPA表达、细胞增殖及细胞迁移率均显著增高(P<0.05),其中LoVo细胞较SW480细胞增高更为显著(P<0.01);(3)与血清组比较,PS-341组与PDTC组中2种细胞总NF-κB、核内NF-κB和uPA表达、细胞增殖及细胞迁移率均显著降低(P<0.05),且LoVo细胞较SW480细胞降低更为显著(P<0.01);(4)与PS-341组比较,PDTC组中2种细胞总NF-κB、核内NF-κB和uPA表达、细胞增殖及细胞迁移均未见明显差别。结论 LoVo细胞较SW480细胞NF-κB、uPA呈高水平表达,其侵袭力高于SW480细胞可能与此有关;LoVo、SW480细胞uPA表达可能受NF-κB调控;PS-341可能通过降低NF-κB、uPA表达抑制LoVo、SW480细胞增殖与迁移;对LoVo细胞侵袭力抑制率高于SW480细胞可能与其对2种细胞NF-κB活性抑制程度不同有关。

Objective To observe the effect of PS-341 on the expression of NF-κB and uPA as well as cell invasiveness in human LoVo,SW480 colon cancer cells.Methods The experiment was divided in 4 groups： control group（serum-free medium）,serum group（10%FCS）,PS-341 group [10%FCS ＋PS-341（4 ng/mL）] and PDTC group [10%FCS ＋PDTC（10-2 mol/L）].The expression of NF-κB and uPA in human LoVo,SW480 cells was determined by Western-blotting.Proliferation and migration were respectively determined by Bromodeoxyuridine（BrdU） ELISA and Boyden chamber.Results（1） Compared with SW480,in serum group,LoVo＇s total NF-κB expression,intranuclear NF-κB expression,uPA expression,proliferation and migration were significantly increased（P〈0.05）（2） Compared with control group,in serum group,LoVo and SW480＇s total NF-κB expression,intranuclear NF-κB expression,uPA expression,proliferation and migration were significantly increased（P〈0.05）,and the former（LoVo） increased more highly（P〈0.01）;（3） Compared with serum group,in PS-341 group and PDTC group,LoVo and SW480＇s total NF-κB expression,intranuclear NF-κB expression,uPA expression,proliferation and migration were significantly decreased（P〈0.05）,and the former（LoVo） decreased more significantly（P〈0.01）;（4） Compared with PS-341 group,In PDTC group,LoVo and SW480＇s total NF-κB expression,intranuclear NF-κB expression,uPA expression,proliferation and migration were not significantly changed.Conclusions LoVo＇s NF-κB and uPA expression is higher than SW480＇s.The higher invasiveness of LoVo than SW480 may be related to the higher expression of NF-κB and uPA in the cell.The uPA expression of LoVo and SW480 may be regulated by NF-κB.PS-341 may inhibit LoVo and SW480＇s proliferation and migration by decreasing the expression of NF-κB and uPA in the cell.The higher invasiveness inhibition ratio by PS-341 in LoVo than SW480 may be ralated to the inhibition lever of NF-κB in the two cells by PS-341.