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HIPK2对肝癌细胞HepG2生物学表型的调控机制研究

Mechanism of regulation for biological characteristics of hepatocellular carcinoma cell line HepG2 by HIPK2
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摘要 目的:研究外源同源结构域相互作用蛋白激酶2(HIPK2)基因表达对人肝癌细胞系HepG2裸鼠成瘤性的影响及机制。方法:将人HIPK2基因的全长cDNA序列克隆至腺病毒载体(pAdeno-HIPK2),制备Adeno-HIPK2重组腺病毒。以50 pfu/细胞的Adeno-HIPK2感染HepG2细胞作为实验组,Adeno-lacZ感染作为阴性对照,未感染细胞作为空白对照,观察各组细胞的裸鼠成瘤性和细胞凋亡情况,并绘制细胞生长曲线。第6周取材,检测各组样品中HIPK2、JNK1、JNK2、P53、mdm2的水平和JNK激酶活性。结果:实验组细胞的裸鼠成瘤性明显降低,细胞凋亡率有所上升。实验组的P53和mdm2的蛋白表达分别是对照组的3倍和15%,JNK1和JNK2基本不变,但JNK激酶活性则升至对照组的5倍。结论:HIPK2高表达可以激活JNK激酶途径,并有效提高细胞内P53水平,使HepG2细胞的裸鼠成瘤性下降,是肝癌生物治疗中重要目的基因。 Objective:To study the mechanism of regulation for nude mouse tumorigenicity of hepatocellular carcinoma cell line HepG2 by homeodomain-interacting protein kinase 2 (HIPK2). Methods:The full-length human HIPK2 cDNA was cloned into adenoviral vectors (pAdeno-HIPK2) and the recombinant Adeno-HIPK2 was amplified in vitro. Infection of HepG2 cells with Adeno-HIPK2 was performed at the ratio of 50 pfu/cell. The HepG2 cells infected with Adeno-lacZ were taken as negative control and non-infected cells as blank control. The tumorigenicity and cell apoptosis in each group were compared. The cell growth curve was painted. Six weeks after implantation into nude mice, the levels of HIPK2, JNK1, JNK2, P53, and mdm2 and the activity of JNK were estimated in various groups. Results:Experimental HepG2 cells indicated lower tumorigenicity and a higher apoptosis rate. Moreover, the production of P53 and mdm2 protein in the experimental group was three times and 15% that of the controls. The expression of JNK1 and JNK2 was stable, however, the activity of JNK in the HepG2 cells with ectopic HIPK2 increased five-fold compared with controls. Conclusion:HIPK2 over-expression may activate the JNK pathway, and increase the level of P53, both of which lead to lower tumorigenicity in HepG2 cells. So it is considered an important target gene in the biological therapy for hepatocellular carcinoma.
出处 《军事医学科学院院刊》 CSCD 北大核心 2008年第1期19-22,26,共5页 Bulletin of the Academy of Military Medical Sciences
关键词 HIPK2 肝癌 JNK丝裂原活化蛋白激酶类 P53 MDM2 细胞凋亡 homeodomain-interacting protein kinase 2 hepatocellular carcinoma JNK mitogen-activated kinases P53 mdm2 apoptosis
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参考文献16

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