宫颈癌中CSN6对E6AP的调节作用及其临床意义

Regulatory Effect of Constitutive Photomorphogenesis 9 Signalosome Subunit 6 on E6AP in Cervical Cancer and Its Clinical Significance

目的:探讨宫颈癌中组成型光形态发生因子9信号复合体亚单位6(constitutive photomorphogenesis 9 signalosome subunit 6,CSN6)对E6AP的调节作用。方法:在癌症和肿瘤基因图谱(The Cancer Genome Atlas,TCGA)数据库中检索CSN6和E6AP基因,比较分析其相关性。采用质粒转染、蛋白质印迹(Western blotting)、四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)、流式细胞术等方法检测CSN6对E6AP表达及细胞生物学的影响。通过在裸鼠皮下接种不同CSN6表达状态的Hela细胞株建立动物模型,观察CSN6对肿瘤生长的影响。结果:数据库资料显示,CSN6和E6AP在肿瘤中有过表达和突变的现象。CSN6可以上调E6AP蛋白的表达,抑制CSN6的表达可以使E6AP表达下降,细胞增殖能力降低,细胞周期停滞于G0-G1期。动物实验显示,敲除CSN6可以显著抑制肿瘤生长,且肿瘤中E6AP蛋白表达水平下降,p53蛋白表达水平增加。结论:CSN6可以通过上调E6AP蛋白的表达而促进宫颈癌的发生发展。

Objective：To explore the role of constitutive photomorphogenesis 9 signalosome subunit 6（CSN6）in the regulation of E6 AP in cervical cancer.Methods：CSN6 and E6 AP genes were retrieved from The Cancer Genome Atlas（TCGA）,and the correlation between them was analyzed.The effect of CSN6 on expression of E6 AP and cytobiology was determined with transfection of plasmids and Western blotting,methyl thiazolyl tetrazolium（MTT）and flow cytometry（FCM）.Nude mice were subcutaneously injected with Hela cell strains with different expression levels of CSN6,so as to set up the animal model.And the effect of CSN6 on growth of tumors was observed.Results：The database showed the overexpression and the mutation of CSN6 and E6AP in cancer.CSN6 could up-regulate the expression of E6 AP.The inhibition on CSN6 could decrease the expression level of E6 AP and the proliferation of cells,and induce the G0-G1 arrest.Animal expreiment showed that knock-out of CSN6 significantly inhibited the tumor growth,decreasing E6 AP level and increasing p53 level.Conclusions：CSN6 could contribute to the occurrence and progression of cervical cancer by up-regulating the expression of E6 AP.