有丝分裂激酶Aurora B对染色体形态变化的影响  

EFFECT OF MITOTIC KINASE AURORA B ON MORPHOLOGICAL CHANGES OF CHROMOSOMES

作　　者：王蕾[1] 宋春林 张雨晴 张刚[1] WANG Lei;SONG Chunlin;ZHANG Yuqing;ZHANG Gang(Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院生物化学与分子生物学系,山东青岛266071

出　　处：《青岛大学学报（医学版）》2023年第1期16-20,共5页Journal of Qingdao University(Medical Sciences)

基　　金：国家自然科学基金资助项目(31970666)。

摘　　要：目的探究有丝分裂激酶Aurora B对HeLa细胞染色体形态变化影响及其可能的作用机制。方法在HeLa细胞中加入二甲基乙酰胺(DMA),观察染色体形态的变化;将HeLa细胞分为A、B组,分别加入DMSO和Aurora B激酶抑制剂处理,应用Western blot检测两组组蛋白H3 S10的磷酸化水平,利用免疫荧光技术、活细胞成像和细胞染色体分离实验检测两组细胞的染色体形态变化;在B组细胞处理的基础上加入磷酸酶抑制剂处理(C组),利用免疫荧光技术检测磷酸酶对染色体形态变化的影响;使用小干扰RNA(siRNA)筛选Aurora B激酶的底物蛋白。结果A组和B组细胞中组蛋白H3 S10磷酸化水平比较差异有显著性(t=23.58,P<0.05)。B组加入Aurora B激酶抑制剂处理30、60、90 min时染色体聚集细胞数量较A组显著增多,差异具有统计学意义(F=379.28~738.73,P<0.05),抑制Aurora B激酶能够促进染色体的聚集。A组、B组、C组染色体聚集的细胞数量差异有显著性(F=969.20,P<0.05),其中C组明显低于B组(t=22.41,P<0.05)。siRNA筛选结果显示,染色体上可能存在多种Aurora B激酶的底物蛋白影响染色体的形态变化。结论有丝分裂激酶Aurora B通过磷酸化多种底物蛋白使染色体处于分散状态,而磷酸酶则发挥去磷酸化作用促进染色体的聚集,两者协同调控有丝分裂染色体的形态变化。Objective To investigate the effect of mitotic kinase Aurora B on chromosome morphological changes in HeLa cells and its possible mechanism.Methods Dimethylacetamide was added to HeLa cells to observe chromosomal morphological changes.HeLa cells were divided into groups A and B and were treated with DMSO and Aurora B inhibitor,respectively.We-stern blot was used to measure the phosphorylation level of histone H3 S10,and immunofluorescence assay,live cell imaging,and cell chromosome segregation experiment were used to observe chromosome morphological changes.The cells treated with phosphatase inhibitor in addition to the treatment in group B were set up as group C,and immunofluorescence assay was used to observe the effect of phosphatase on chromosome morphological changes.Small interfering RNA(siRNA)was used to screen out the substrate proteins of Aurora B.Results There was a significant difference in the phosphorylation level of histone H3 S10 between groups A and B(t=23.58,P<0.05).Compared with group A,group B had a significant increase in the number of chromosome aggregation cells at 30,60,and 90 minutes of Aurora B inhibitor treatment(F=379.28-738.73,P<0.05),indicating that inhibition of Aurora B promoted chromosome aggregation.There was a significant difference in the number of chromosome aggregation cells between groups A,B,and C(F=969.20,P<0.05),and group C had a significantly lower number than group B(t=22.41,P<0.05).The results of siRNA screening showed that there might be multiple Aurora B substrate proteins on chromosomes that in-fluenced chromosome morphological changes.Conclusion The mitotic kinase Aurora B makes the chromosomes in a dispersed state by phosphorylating multiple substrate proteins,while phosphatase exerts a dephosphorylation effect to promote the aggregation of chromosomes,thereby regulating the morphological changes of mitotic chromosomes in a synergistic way.

关 键 词：极光激酶B 有丝分裂 染色体 

分 类 号：R345.57[医药卫生—基础医学]