沉默wee1基因对卵巢癌细胞增殖、迁移和侵袭的作用机制

目的探讨沉默wee1基因对卵巢癌细胞增殖、迁移和侵袭的作用机制。方法将卵巢癌细胞分为Blank组、小干扰RNA(siRNA)阴性对照(siRNA-NC)组和携带wee1基因片段(siRNA-wee1)组,利用siRNA技术在卵巢癌细胞中沉默wee1基因,采用实时荧光定量PCR、免疫印迹法(WB)检测3组卵巢癌细胞wee1 RNA及蛋白表达水平,采用细胞划痕试验、Transwell试验观察卵巢癌细胞平板集落形成数及卵巢癌细胞迁移、侵袭能力,采用WB观察卵巢癌细胞Wnt/β-catenin信号通路相关蛋白Wnt3a、β-连环蛋白(β-catenin)、细胞周期蛋白D1(Cyclin-D1)、基质金属蛋白酶7(MMP-7)的表达情况。结果siRNA-wee1组卵巢癌细胞wee1 RNA和蛋白表达水平均低于siRNA-NC组和Blank组(P<0.05),卵巢癌细胞集落形成数少于Blank组和siRNA-NC组(P<0.05),卵巢癌细胞迁移、侵袭能力低于Blank组和siRNA-NC组(P<0.05),Wnt/β-catenin信号通路相关蛋白Wnt3a、β-catenin、Cyclin-D1和MMP-7表达水平均低于siRNA-NC组和Blank组(P<0.05)。结论沉默wee1基因可抑制Wnt/β-catenin信号通路的激活,进而抑制卵巢癌细胞增殖、迁移和侵袭能力。

Mechanism of silencing wee1 gene on proliferation,migration and invasion of ovarian cancer cells

Objective To investigate the mechanism of silencing wee1 gene on proliferation,migration and invasion of ovarian cancer cells.Methods The ovarian cancer cells were divided into Blank group,small interfering RNA(siRNA)negative control(siRNA-NC)group,and carrying wee1 gene fragment(siRNA-wee1)group.The wee1 gene was silenced in ovarian cancer cells by siRNA technology.The expression levels of wee1 RNA and protein in 3 groups of ovarian cancer cells were detected by real-time fluorescence quantitative PCR and Western blotting(WB).The cell scratch test and the Transwell test were used to observe the number of ovarian cancer cell plate colonies and the migration and invasion capabilities of ovarian cancer cells.WB was used to observe the expression of Wnt3a,β-catenin,Cyclin-D1 and matrix metalloproteinase 7(MMP-7)related to the Wnt/β-catenin signaling pathway in ovarian cancer cells.Results The expression levels of wee1 RNA and protein in ovarian cancer cells in the siRNA-wee1 group were lower than those in the siRNA-NC group and the Blank group(P<0.05),and the number of colonies formed in ovarian cancer cells was less than that of the Blank group and the siRNA-NC group(P<0.05).The migration and invasion ability of ovarian cancer cells was lower than those of Blank group and siRNA-NC group(P<0.05).The expression levels of Wnt/β-catenin signaling pathway related proteins Wnt3a,β-catenin,Cyclin-D1 and MMP-7 in the siRNA-wee1 group were lower than those in the siRNA-NC group and Blank group(P<0.05).Conclusion Silencing the wee1 gene can inhibit the activation of Wnt/β-catenin signaling pathway,thereby inhibiting the proliferation,migration and invasion of ovarian cancer cells.