YB-1对卵巢癌细胞EMT标志蛋白表达及黏附能力的影响研究

Effect of YB-1 on expression and adhesion ability of EMT marker protein in ovarian cancer cells

目的研究Y-box结合蛋白(YB-1)对卵巢癌细胞上皮间质转化(EMT)标志蛋白表达及细胞黏附能力的影响。方法在卵巢癌细胞中转染p Genesil-1.1/YB-1 shRNA和p Genesil-1.1/shRNA control,筛选稳定转染的细胞记为YB-1 shRNA组和shRNA-NC组,把没有转染的细胞作为对照组。以Realtime PCR和Western blot方法检测细胞中YB-1表达水平。用四甲基偶氮唑盐比色法(MTT)方法分别检测细胞增殖和黏附能力,用Western blot检测EMT标志蛋白上皮钙黏附素(E-cadherin)、神经型钙粘素(N-cadherin)和迁移侵袭相关蛋白基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)水平,Transwell小室测定细胞侵袭和迁移情况。结果 YB-1 shRNA组细胞中的YB-1表达水平明显低于对照组和shRNA-NC组(P<0.05)。YB-1 shRNA组细胞增殖率和黏附率低于对照组和shRNA-NC组(P<0.05)。YB-1 shRNA组细胞中E-cadherin蛋白水平高于对照组和shRNA-NC组,N-cadherin、MMP-2、MMP-9蛋白水平低于对照组和shRNA-NC组,差异具有统计学意义(P<0.05)。YB-1 shRNA组细胞侵袭和迁移数目低于对照组和shRNA-NC组(P<0.05)。结论下调YB-1促进卵巢癌细胞E-cadherin表达,抑制N-cadherin表达,抑制细胞EMT,降低细胞黏附、侵袭、迁移能力。

Objective To study the effect of YB-1 on the expression of EMT marker protein and cell adhesion in ovarian cancer cells.Methods PGenesil-1. 1/YB-1 shRNA and p Genesil-1. 1/shRNA control were transfected into ovarian cancer cells,and stable transfected cells were identified as YB-1 shRNA group and shRNA-NC group,the cells that were not transfected were used as Control group. The expression level of YB-1 in cells was detected by real-time PCR and Western blot,cell proliferation and adhesion ability were detected by MTT,Western blot was used to detect EMT protein E-cadherin,N-cadherin,migration and invasion related protein MMP-2 and MMP-9 levels,cell invasion and migration were measured in Transwell chamber. Results The expression level of YB-1 in YB-1 shRNA group was significantly lower than that in group Control and shRNA-NC（P〈0. 05）. The cell proliferation rate and adhesion rate of YB-1 shRNA group were lower than those of Control group and shRNA-NC group（P〈0. 05）. The level of E-cadherin protein in group YB-1 shRNA was higher than that in group Control and shRNA-NC,the protein levels of N-cadherin,MMP-2 and MMP-9 were lower than those in group Control and shRNA-NC,the difference was statistically significant（P〈0. 05）. The number of invasion and migration in YB-1 shRNA group was lower than that in Control group and shRNA-NC group（P〈0. 05）. Conclusion Down regulation of YB-1 can promote the expression of E-cadherin,inhibition of N-cadherin expression,inhibit EMT,reduce cell adhesion,invasion and migration ability of ovarian cancer cells.