结直肠癌组织和SW480细胞系中SKA3蛋白的表达及对PI3K/AKT信号通路的影响

Expression of SKA3 protein in colorectal cancer tissues and SW480 cell lines and its effect on PI3K/AKT signaling pathway

目的探讨结直肠癌组织中SKA3的表达及临床意义,观察转染SKA3-siRNA对结直肠癌SW480细胞增殖、迁徙的影响及在PI3K/AKT信号通路中的作用。方法采用免疫组化法观察69例结直肠癌和癌旁组织中SKA3蛋白的表达水平。结直肠癌SW480细胞转染SKA3-siRNA干扰后,应用Western blot法观察细胞中SKA3蛋白的表达;MTT法和细胞克隆实验观察细胞增殖改变,细胞划痕法观察细胞迁徙改变,Western blot法检测PI3K/AKT信号通路蛋白在细胞中的表达。结果结直肠癌组织中的SKA3表达水平比癌旁组织显著增加(P<0.01),且SKA3蛋白表达水平与结直肠癌分化程度、淋巴结转移等有关(P<0.05);结直肠癌SW480细胞转染SKA3-siRNA后,SKA3表达水平明显降低(P<0.05);同时,结直肠癌SW480细胞增殖能力受抑,迁徙能力降低(P<0.05);p-AKT、p-PI3K蛋白水平显著下降(P<0.05)。结论SKA3在结直肠癌中呈高表达,与患者预后相关;敲减SKA3表达可抑制结直肠癌SW480细胞的增殖和迁徙,其机制可能与抑制PI3K/AKT信号通路有关。

Purpose To investigate the expression of spindle and kinetochore associated complex subunit 3(SKA3)protein in colorectal cancer tissues and its clinical significance,and to observe the effect of SKA3-siRNA on the proliferation and migration of colorectal cancer SW480 cells and its role of in PI3K/AKT signaling pathway.Methods The expression level of SKA3 in 69 cases of colorectal cancer and paracancerous tissue was observed by immunohistochemistry.After SKA3-siRNA gene transfection,the expression of SKA3-siRNA in colorectal cancer SW480 cells was observed by Western blot.MTT assay and clonal formation assay were used to observe the proliferation of colorectal cancer cells after transfection,and cell scratch assay was used to observe the migration of SW480 cells.Western blot was used to analyze the mechanism of the influence of PI3K/AKT signaling pathway on SKA3-siRNA in colorectal cancer SW480 cells.Results SKA3 expression was significantly increased in colorectal cancer tissues compared with paracancerous tissues(P<0.01),and the expression level of SKA3 protein was related to the degree of differentiation and lymph node metastasis of colorectal cancer(P<0.05).After SKA3-siRNA in colorectal cancer SW480 cells,the expression level of SKA3 in colorectal cancer SW480 cells was significantly decreased(P<0.05),meanwhile,proliferation and migration of colorectal cancer SW480 cells were inhibited(P<0.05),SKA3-siRNA could activate the PI3K/AKT signaling pathway(P<0.05).Conclusion SKA3 is highly expressed in colorectal cancer and is related to the prognosis of patients.SKA3-siRNA can inhibit the proliferation and migration of colorectal cancer SW480 cells,and SKA3 may become a molecular marker to predict the development and prognosis of colorectal cancer patients.