SKA3促进子宫内膜癌细胞增殖、迁移和侵袭

SKA3 Promotes the Proliferation,Migration and Invasion of Endometrial Carcinoma Cells

目的探讨纺锤体与着丝粒相关蛋白3(SKA3)对子宫内膜癌细胞增殖、迁移和侵袭的影响。方法通过RT-qPCR和蛋白质印迹技术检测子宫内膜上皮细胞HEECs以及子宫内膜癌细胞KLE、JEC、Ishikawa细胞中SKA3 mRNA水平和蛋白表达,选取表达量最高的Ishikawa细胞用于后续研究。将Ishikawa细胞分为3组:对照组:正常培养Ishikawa细胞;si-SKA3组:根据脂质体2000说明书,将50 nmol/L siRNA-SKA3转染至Ishikawa细胞;NC组:根据脂质体2000说明书,将50 nmol/L siRNA-NC转染至Ishikawa细胞。通过CCK8和克隆形成实验检测3组细胞增殖情况,Transwell和划痕实验检测3组细胞侵袭和迁移情况,蛋白质印迹技术检测3组细胞CyclinD1、MMP-2、AKT、p-AKT蛋白表达。结果与子宫内膜上皮细胞HEECs比,各子宫内膜癌细胞系中SKA3 mRNA和蛋白表达量均升高(P<0.05)。干扰SKA3表达后,Ishikawa细胞增殖能力、侵袭能力降低,划痕宽度变大,CyclinD1和MMP-2蛋白表达量均下降(P<0.05)。与对照组和NC组比,si-SKA3组细胞AKT蛋白表达无明显变化(P>0.05),p-AKT蛋白表达下降(P<0.05)。结论干扰SKA3能够抑制子宫内膜癌细胞Ishikawa增殖、迁移和侵袭,其机制可能与SKA3抑制PI3K/AKT信号通路有关。

Objective To investigate the effects of spindle and kinetochore associated complex subunit 3(SKA3)on the proliferation,migration and invasion of endometrial carcinoma cells.Methods Real-time quantitative polymerasechain reaction(RT-qPCR)and Western blotting were used to detect the levels of SKA3 mRNA and protein expression in endometrial epithelial cells HEECs and endometrial carcinoma cells KLE,JEC and Ishikawa cells.And the Ishikawa cells with the highest expression level were selected for subsequent research.Ishikawa cells were divided into three groups.In the control group,Ishikawa cells were cultured normally.In the si-SKA3 group,50 nmol/L siRNA-SKA3 was transfected into Ishikawa cells according to liposome 2000 instructions.In the NC group,50 nmol/L siRNA-NC was transfected into Ishikawa cells according to liposome 2000 instructions.Cell proliferation was detected by cell counting kit-8(CCK8)and clonal formation assay.Cell invasion and migration were detected by Transwell and scratch assay.And protein expression of CyclinD1,MMP-2,AKT,and p-AKT were detected by Western blotting.Results Compared with the HEECs of endometrial epithelial cells,the expression levels of SKA3 mRNA and protein in each endometrial carcinoma cell line were significantly increased(P<0.05).After interference with SKA3 expression,the proliferation and invasion ability of Ishikawa cells were decreased,the width of scratches were enlarged,and the protein expression of CyclinD1 and MMP-2 were significantly decreased(P<0.05).Compared with the control group and the NC group,the expression of AKT protein in si-SKA3 group was not significantly changed(P>0.05),while the expression of p-AKT protein decreased(P<0.05).Conclusion Interference with SKA3 can inhibit the proliferation,migration and invasion of Ishikawa cells of endometrial carcinoma,of which the mechanism may be related to the inhibition of SKA3 on PI3K/AKT signaling pathway.