细胞质唾液酸酶(Neu2)通过Cdk2通路抑制FBJ-LL细胞生长  

Induction of G1 cycle arrest by over-expression of neuraminidase 2 in FBJ-LL cell possibly via down-regulation of Cdk2 mRNA

作　　者：陈阳[1] 吴英良[1] 山形贞子[1] 山形达也[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2010年第2期147-151,共5页Journal of Shenyang Pharmaceutical University

摘　　要：目的研究细胞质唾液酸酶(neuraminidase 2,Neu2)调控FBJ-LL细胞生长的作用机制。方法利用逆转录聚合酶链式反应(reverse transcriptional-polymerase chain reaction,RT-PCR)检测细胞内Neu2及周期蛋白依赖性激酶2(cyclin dependent kinase 2,Cdk2)的mRNA表达水平,采用酶活性检测法检测细胞内总唾液酸酶的活性,细胞活性法检测细胞的生长情况,碘化丙啶染色法辅助流式细胞仪分析处于不同细胞周期的DNA含量分布。结果Neu2 sense cDNA转染的FBJ-LL细胞中Neu2 mRNA的表达显著升高,其总唾液酸酶的活性也随之升高。FBJ病毒诱导的小鼠骨肉瘤(FBJ-LL)细胞中Neu2的过量表达可以抑制该细胞的生长,升高该细胞中处于细胞周期G1期细胞的相对数量,同时抑制了该细胞中的Cdk2的mRNA表达水平。结论Neu2通过抑制细胞周期分子Cdk2的表达来阻碍FBJ-LL细胞DNA合成,从而实现对FBJ-LL细胞的生长抑制。Objective To verify the roles of Neu2 （neuraminidase） as a growth regulator in FBJ-LL cells.Methods The mRNA expression of Neu2 and Cdk2 were determined by reverse transcription-polymerase chain reaction. The sialidase activity was determined by using 4-MUNeuAC as a substrate. Cell viability was detected by tetrazolium assay. Cell cycle analysis was performed by propidium iodide staining followed by flow cytometry. Results Neu2 sense cDNA transfection significantly enhanced the mRNA expression of Neu2 and the total sialidase activity in FBJ-LL cells. Neu2 over-expression in FBJ-LL cells inhibited the cell growth and decreased the number of cells in G1 phase. Neu2 sense cDNA transfected cells showed impaired expression of Cdk2 mRNA. Conclusions Neu2 is responsible for cell growth inhibition of FBJ-LL cellsthrough Cdk2 signal pathways.

关 键 词：细胞质唾液酸酶2 周期蛋白依赖性激酶2 FBJ细胞 细胞生长抑制 细胞周期G1 

分 类 号：R963[医药卫生—微生物与生化药学]