泛素特异性蛋白酶4对透明细胞肾细胞癌细胞增殖能力的影响

Effect of ubiquitin specific protease 4 on cell proliferation of kidney renal clear cell carcinoma

目的探讨人类泛素特异性蛋白酶4(USP4)对透明细胞肾细胞癌(KIRC)细胞增殖能力的影响。方法从TCGA数据库及GEO数据库中下载KIRC患者的基因表达及临床信息资料,进行转化及整理;同时利用Ualcan、HPA数据库等在线工具对数据库中的KIRC患者进行进一步分析,研究USP4表达与KIRC患者临床病理特征(包括TNM分期和病理分级)以及生存预后的关系。在KIRC细胞株Caki-1、OSRC-2中转染USP4干扰和过表达质粒,通过平板克隆、细胞计数试剂盒8实验检测USP4对KIRC细胞增殖能力的影响。结果生物信息学分析显示,USP4在KIRC组织中的mRNA及蛋白表达水平均明显低于癌旁正常肾脏组织(均P<0.05)。KIRC组织中USP4 mRNA表达水平与患者肿瘤T分期、病理分期、组织学分级、总生存率、疾病特异性生存率、无进展生存率均有关(均P<0.05);与性别、年龄、N分期、M分期均无关(均P>0.05)。细胞实验结果显示,USP4蛋白在Caki-1、OSRC-2细胞中显著过表达,而过表达USP4会明显降低Caki-1、OSRC-2细胞增殖能力和克隆形成能力(均P<0.05);敲低USP4会明显提高Caki-1细胞克隆形成能力(均P<0.05)。结论USP4在KIRC组织中低表达,而过表达USP4可抑制KIRC细胞增殖能力。

Objective To explore the effect of human ubiquitin specific protease 4(USP4)on cell proliferation of kidney renal clear cell carcinoma(KIRC).Methods The gene expression and clinical information of KIRC patients were obtained from TCGA database and GEO database.The relationship of USP4 expression with clinicopathological features and of KIRC and survival of patients were analyzed with online tools Ualcan and HPA database.Human KIRC Caki-1 and OSRC-2 cells were transfected with USP4 overexpression or interference plasmids,respectively.The effects of overexpression or knockdown of USP4 on the proliferation ability,cloning ability of KIRC cells were detected by cell counting kit-8 method and plate cloning assay,respectively.Results Bioinformatics analysis showed that the expression levels of USP4 mRNA and protein in KIRC tissues was lower than that in corresponding adjacent tissues(all P<0.05).The expression of USP4 protein in KIRC tissues was correlated with T stage,pathological grade,histological grade,overall survival,disease specific survival and progression-free survival(all P<0.05),not correlated with gender,age,N stage,M stage of KIRC patients(all P>0.05).Cell experiment results showed that USP4 protein was significantly overexpressed in Caki-1 and OSRC-2 cells,and overexpression of USP4 significantly decreased the proliferation and clonogenesis ability of Caki-1 and OSRC-2 cells(both P<0.05).Knockdown of USP4 significantly improved the cloning ability of Caki-1 cells(all P<0.05).Conclusion USP4 is underexpressed in KIRC,and overexpression of USP4 can inhibit the proliferation of KIRC cells.