BUB1B在原发性肝癌中的表达及对肝癌细胞增殖和侵袭的影响

Clinical significance of BUB1B in hepatocellular carcinomaand its effect on proliferation and invasion of hepatoma cells

目的探究纺锤体检测蛋白BUB1B在原发性肝癌(HCC)患者中的表达及临床意义,并研究其在体外对肝癌细胞增殖和侵袭的影响。方法收集GEO数据库中HCC患者(GSE121248)的临床以及基因表达信息,通过生物信息学分析筛选差异表达基因,设计针对BUB1B基因的小干扰RNA载体,通过沉默BUB1B基因表达验证其在肝癌细胞增殖以及侵袭中的影响。利用实时荧光定量PCR检测沉默效率,通过细胞增殖实验以及划痕实验分别检测该基因在肝癌细胞增殖以及侵袭中的作用。利用KM-plotter数据库分析BUB1B基因表达高低对HCC患者预后的影响。结果沉默序列1和2能够有效地沉默BUB1B基因的mRNA水平表达(t=8.41,P=0.001;t=10.43, P <0.001),沉默BUB1B表达的肝癌细胞系HepG2的增殖能力减弱(t=13.94,P <0.001),肝癌细胞8 h的侵袭能力同样减小(t=14.94,P <0.001)。高表达BUB1B基因的HCC患者(n=127)生存率差于低表达BUB1B基因的HCC患者(n=237)[HR=2.01(1.42～2.86),P=6.6×10^(-5)],进一步分析发现,在有肝炎病毒感染史的HCC患者(n=150)中,BUB1B基因表达高低对预后没有影响[HR=1.88(0.89～3.99), P=0.093],只有在未感染肝炎病毒的HCC患者(n=167)中可以作为预测预后的分子标志物[HR=2.84(1.75～4.61), P=1.1×10^(-5)]。结论BUB1B基因在HCC患者中高表达,促进肝癌细胞增殖以及侵袭,在未感染肝炎病毒的HCC患者中,可以作为预测预后的潜在分子标志物。

Objective To investigate the expression and clinical significance of BUB1B in patients with hepatocellular carcinomaan(d HCC), and to evaluate its effect on the proliferation and invasion of human liver cancer cells in vitro. Methods The clinical data and the information of gene expression were collected from GEO database (GSE121248). The differentially-expressing genes were screened and analyzed via bioinformatics analysis. A small interfering RNA vector wa sdesigned targeting BUB1B. The effect of BUB1B gene on the proliferation and invasion of liver cancer cells was assessed by silencing the expression of BUB1B gene. The knockdown efficiency was detected by quantitative real-time PCR. The proliferating and invasion abilities were evaluated by cell proliferation assay and wound healing test. The effect of the expression levels of BUB1B gene upon the clinical prognosis of HCC patienst was assessed by KM-plotter database. Results The silence sequences 1 and 2 could significantly silence the xepression of BUB1B mRNA( t=8.41, P=0.001;t=10.43, P < 0.001). The proliferating ability of hepatoma cell line HepG2 ,which knocked down the expression of BUB1B gene, was significantly weakened( t=13.94, P < 0.001). The invasion ability was also reduced at 8 h after knocking down the expression of BUB1 Bgene( t=14.94, P < 0.001). The survival rate of HCC patients with high expression of BUB1B ge(ne n=127) was significantly lower than that of their counterparts with low expression of BUB1B gen(e n = 237)[ HR = 2.01( 1.42 ~ 2.86), P = 6.6×10-5]. Further analysis found that the expression of BUB1B gene exerted no effect on the clinical prognosis of HCC patient s with a medical history of hepatitis( n = 150)[ HR = 1.88( 0.89 ~ 3.99), P=0.093], whereas it acted as a molecular biomarker to predict the clinical prognosis of HCC tpiaents without a medical history of hepatiti s (n = 167)[HR = 2.84 (1.75 ~ 4.61), P = 1.1×10^-5]. Conclusions The BUB1B gene is highly expressed in HCC patients, which can promote the proliferation and invasion of liver caenrc cells. BUB1B gene is a potentia l molecular biomarker for predicting the clinical prognosis of HCC Cpatients with no medical history of hepatitis.