细胞周期依赖性蛋白激酶对体外培养人肝癌细胞侵袭力的影响  

Cyclin-dependent kinase inhibitor roscovitine suppresses the invasion of human hepatocellular carcinoma SMMC-7721 cells in vitro

作　　者：王开阳[1] 熊爱珍[2] 蒋星星[1] 李龙[1] 李慧珍[1] 傅华群[1] 

机构地区：[1]南昌大学第二附属医院肝胆外科,江西省南昌市330006 [2]南昌大学第二附属医院药剂科,江西省南昌市330006

出　　处：《世界华人消化杂志》2010年第2期119-124,共6页World Chinese Journal of Digestology

基　　金：江西省卫生厅基金资助项目;No.20091156~~

摘　　要：目的:探讨CDKs对体外培养的人肝癌细胞SMMC-7721侵袭力的影响及其分子机制.方法:将细胞分为A、B两组(A组用终浓度为0μmol/L Roscovitine,B组用终浓度为32μmol/L Roscovitine,均培养24h),采用流式细胞术检测经CDKs特异性抑制剂Roscovitine干预后的人肝癌细胞SMMC-7721的细胞周期,并用Transwell小室、划痕实验、PCR技术分别检测处于不同细胞周期下的人肝癌细胞侵袭能力、水平运动能力及uPA、MMP-9mRNA表达.结果:经终浓度为32μmol/L的Roscovitine干预24h后的人肝癌细胞SMMC-7721处于G0/G1期细胞比例迅速升高(72.19%±0.47%vs59.22%±0.54%,P<0.05),细胞侵袭能力下降,穿膜细胞数明显减(71.40±5.59vs149.60±16.36,P<0.05);细胞水平运动能力明显下降(P<0.05);uPAmRNA的表达下降、但MMP-9mRNA的表达却无明显变化.结论:Roscovitine干预使人肝癌细胞SMMC-7721侵袭能力和水平运动能力下降,其机制可能与肝癌细胞周期时相分布发生改变和uPAmRNA的表达下降有关.AIM：To investigate the effects of cyclindependent kinases on the invasion of human hepatocellular carcinoma SMMC-7721 cells in vitro and explore potential mechanisms involved.METHODS：SMMC-7721 cells were divided into two groups：control group （untreated with roscovitine） and treatment group （treated with 32 μmol/L of roscovitine for 24 hours）.The ell cycle distribution of SMMC-7721 cells was detected by flow cytometry.Cell invasion and motility were evaluated by Transwell chamber assay and wound healing assay,respectively.The mRNA expression of urokinase plasminogen activator （uPA） and matrix metalloproteinase 9 （MMP-9） was detected by reverse transcriptionpolymerase chain reaction （RT-PCR）.RESULTS：The percentage of SMMC-7721 cells in G 0 /G 1 phase was significantly higher in the treatment group than in the control group （72.19% ± 0.47% vs 59.22% ± 0.54%,P 0.05）.The number of cells passing through the Transwell membrane was significantly lower in the treatment group than in the control group （71.40 ± 5.59 vs 149.60 ± 16.36,P〈 0.05）.Roscovitine treatment also significantly decreased cell motility （P 〈0.05）.RT-PCR analysis revealed that roscovitine treatment downregulated the expression of uPA mRNA expression but had no significant impact on MMP-9 mRNA expression.CONCLUSION：Roscovitine treatment decreases the invasion and motility of SMMC-7721 cells possibly via a mechanism associated with changing cell cycle and downregulating uPA mRNA expression.

关 键 词：细胞周期依赖性蛋白激酶 细胞周期 侵袭力 UPA 肝癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]