下调HOXA5表达对胶质瘤细胞增殖能力和细胞周期的影响  

Effect of HOXA5 knockdown on proliferation and cell cycle of human glioma cells

作　　者：徐修鹏[1] 季晶[1] 刘宁[1] 李海林[1] 路华[1] XU Xiu-peng;JI Jing;LIU Ning;LI Hai-lin;LU Hua(Department of Neurosurgery,The First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院神经外科,南京210029

出　　处：《中国临床神经外科杂志》2021年第11期857-861,共5页Chinese Journal of Clinical Neurosurgery

基　　金：江苏省自然科学基金(BK20201077)。

摘　　要：目的探讨下调同源异型盒基因A5(HOXA5)表达对胶质瘤细胞增殖能力和细胞周期的影响。方法应用生信分析方法,计算机检索CGGA数据库,下载m RNAseq-693和m RNAseq-325芯片数据,获取低级别胶质瘤(WHO分级Ⅱ级)和高级别胶质瘤(WHO分级Ⅲ、Ⅳ级)HOXA5 m RNA表达及病人生存信息;以HOXA5表达水平的平均值为界限,将高级别胶质瘤分为低表达组和高表达组。体外培养人胶质瘤细胞株U87和U251,使用Lipofectamine 2000法将HOXA5 si RNAs和阴性对照si RNAs分别转染U87和U251细胞;CCK-8和平板克隆形成实验评估胶质瘤细胞的增殖能力;流式细胞仪检测细胞周期;免疫印迹法检测蛋白表达水平。结果生信分析结果显示:高级别胶质瘤HOXA5表达水平均显著高于低级别胶质瘤(P<0.01);HOXA5高表达组高级别胶质瘤病人中位生存时间较低表达组明显缩短(P<0.01)。体外细胞实验结果:敲低HOXA5表达,U87和U251细胞的增殖率和克隆集落数量显著降低(P<0.01),U87和U251细胞G0/G1期细胞百分比显著增高(P<0.01),U87和U251细胞CDK4蛋白表达水平显著降低(P<0.01)。结论胶质瘤HOXA5呈高表达,病理级别越高,表达水平越高,病人生存预后越差。敲低HOXA5表达,明显降低CDK4表达,使细胞周期停滞在G0期,明显抑制胶质瘤细胞的增殖。Objective To explore the effect of homeobox A5(HOXA5)knockdown on the proliferation and cell cycle of glioma cells.Methods The CGGA database was searched to download the chip data of m RNAseq-693 and m RNAseq-325,including lowgrade glioma(WHO gradeⅡ)and high-grade glioma(WHO gradeⅢ~Ⅳ)HOXA5 m RNA expression and patient survival data;the high-grade glioma patients were divided into low-expression group and high-expression group according to the average level of HOXA5 expression.Human glioma cell lines U87 and U251 were cultured in vitro,and HOXA5 si RNAs and negative control si RNAs were transfected into U87 and U251 cells by the Lipofectamine 2000 method,respectively.CCK-8 and clone formation assay were used to evaluate the proliferation ability of glioma cells.Flow cytometry assay was used to detect the cell cycle.Western blotting was used to detect protein expression level of CDK4.Results The results of bioinformatics analysis showed:the expression level of HOXA5 in highgrade gliomas was significantly higher than that in low-grade gliomas(P<0.01);the median survival time of patients with high-grade gliomas in high-expression group was significantly shorter than that in the low-expression group(P<0.01).In vitro cell cultured experiment results showed:knockdown of HOXA5 significantly reduced the proliferation rate and the number of clone colonies of U87 and U251 cells(P<0.01),significantly increased the percentage of cells in the G0/G1 phase of U87 and U251 cells(P<0.01),significantly reduced the expression level of CDK4 of U87 and U251 cells(P<0.01).Conclusions HOXA5 is highly expressed in gliomas.The higher the pathological grade,the higher the expression level of HOXA5,the worse the patient’s survival prognosis.Knockdown of HOXA5 significantly reduce the expression of CDK4,arrest the cell cycle in the G0 phase,and significantly inhibit the proliferation of glioma cells.

关 键 词：胶质瘤 HOXA5 细胞增殖 细胞周期 CDK4 基因敲减 

分 类 号：R739.41[医药卫生—肿瘤] Q786[医药卫生—临床医学]