摘要
目的研究siRNA靶向干扰轴突导向受体蛋白(ROBO)1基因表达对食管癌EC109细胞增殖、细胞周期的影响及可能作用机制。方法实验将食管癌EC109细胞随机分为空白组、对照组、siRNA-ROBO1组,采用脂质体将阴性siRNA和siRNAROBO1转染EC109细胞;Western印迹检测ROBO1、细胞核相关抗原Ki-67(Ki-67)、增殖细胞核抗原(PCNA)、细胞周期蛋白(Cyclin)D1蛋白的表达;CCK-8实验检测细胞的增殖活力;流式细胞仪检测EC109细胞周期的变化。结果干扰ROBO1表达后,EC109细胞中ROBO1蛋白的表达量较空白组显著降低(P<0.05),细胞增殖活性显著降低(P<0.05),G0/G1期细胞显著增加(P<0.05),G2/M期细胞显著降低(P<0.05),且siRNA-ROBO1组细胞中Ki-67、PCNA、Cyclin D1表达量较空白组明显降低(P<0.05)。结论ROBO1可能通过影响食管癌细胞增殖、周期相关蛋白水平使细胞周期阻滞在G0/G1期,从而抑制食管癌细胞增殖。
Objective To investigate the effect of siRNA targeting interference roundabout,axon guidance receptor,homoloh 1（ ROBO1） gene expression on proliferation and cell cycle of esophageal cancer EC109 cells and its possible mechanism. Methods Esophageal cancer EC109 cells were randomly divided into blank group,control group and SiRNA-ROBO1 group. Negative siRNA and siRNA-ROBO1 were transfected into EC109 cells by liposome. The expression of ROBO1,nuclear related antigen Ki-67（ Ki-67）,proliferating cell nuclear antigen（ PCNA） and Cyclin D1 protein were detected by Western blot. The proliferation activity of the cells was detected by CCK-8 assay and the cell cycle changes of EC109 were detected by flow cytometry. Results After interfering with ROBO1 expression,the expression of ROBO1 protein in EC109 cells was significantly lower than that in the blank group（ P〈0.05）,and the cell proliferation activity was significantly decreased（ P〈0.05）. The cells in G0/G1 phase were significantly increased（ P〈0. 05）,and in G2/M phase were significantly decreased（ P〈0.05）. The expression levels of Ki-67,PCNA and Cyclin D1 in siRNA-ROBO1 group were significantly lower than those in the blank group（ P〈0.05）.Conclusions ROBO1 may inhibit the proliferation of esophageal cancer cells by affecting the proliferation and cycleassociated protein levels of esophageal cancer cells and arresting the cell cycle in G0/G1 phase.
作者
刘志广
刘芬
韩江红
LIU Zhi-Guang;LIU Fen;HAN Jiang-Hong(No.1 Department of Chest Tumor,Xinxiang Central Hospital,Xinxiang 453000,Henan,China)
出处
《中国老年学杂志》
CAS
北大核心
2018年第17期4216-4219,共4页
Chinese Journal of Gerontology
基金
国家自然科学基金青年项目(81501991)
