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Cdk1在HepG2细胞的表达及活性对肝癌细胞凋亡的影响

, PP. 521-524

Keywords: Cdk1,siRNA,细胞凋亡,激酶活性

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Abstract:

目的观察Cdk1蛋白表达及其活性对肝癌细胞凋亡的影响。方法克隆Cdk1基因质粒和Cdk1siRNA,并将其转染到肝癌HepG2细胞中,另外应用roscovintine(Cdks的抑制剂)刺激细胞,然后给予紫外线(UV)照射。用WesternBlot检测转染的Cdk1蛋白的表达,用核素标记测定其活性;用HO33342(Hoechst33342)的免疫荧光染色细胞核判断细胞凋亡的情况。分析Cdk1蛋白表达及活性对肝癌细胞凋亡的影响。结果Cdk1可以在HepG2良好地表达;Cdk1siRNA可以有效地沉默Cdk1的表达;roscovintine可抑制Cdk1的活性;细胞凋亡率在roscovintine刺激细胞时最高比较Cdk1转染细胞组和Cdk1siRNA转染细胞组(P<0.01),在Cdk1转染细胞高于Cdk1siRNA转染细胞组(P<0.01)和空载体转染细胞组(P<0.05),在Cdk1siRNA转染细胞最低。结论过度表达Cdk1可以增加肝癌细胞死亡,抑制肝癌的增殖。抑制Cdk1活性可更有效增加肝癌细胞死亡,抑制肝癌的增殖,可望成为研究和探讨肝癌靶点之一。

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