Skp2基因表达下调对HepG2细胞生物学特性及SP1和p27蛋白表达的影响  

Effect of down-regulation of Skp2 gene expression on biological characters of HepG2 cells as well as expressions of SP1 and p27 proteins

作　　者：唐卫军 张幸平[2] 邹洪元 彭春芳 

机构地区：[1]重庆市江津区中心医院肿瘤科,重庆402260 [2]重庆医科大学附属第一医院肿瘤科,重庆400016

出　　处：《中国生物制品学杂志》2012年第1期18-22,28,共6页Chinese Journal of Biologicals

摘　　要：目的研究Skp2基因表达下调对HepG2细胞生物学特性及p27和SP1蛋白表达的影响。方法构建Skp2基因干扰慢病毒质粒,包装后感染HepG2细胞,48 h后,流式细胞仪检测细胞周期和凋亡情况;侵袭小室试验检测细胞侵袭能力;Western blot检测Skp2基因下调后p27和SP1蛋白的表达情况。结果与正常HepG2细胞组相比,Skp2干扰慢病毒组细胞Skp2蛋白的表达明显下调,p27蛋白表达大幅增加,细胞生长速度明显减慢,细胞凋亡率增加近两倍,G0/G1期细胞增多;SP1蛋白表达减少,浸润、迁移的细胞数减少约50%(P<0.05)。结论 Skp2蛋白表达下调抑制了HepG2细胞的增殖,促进HepG2细胞的凋亡,造成细胞侵袭能力减弱。Skp2蛋白表达下调导致p27蛋白表达上调、SP1蛋白表达下调,在上述生物学特性变化中可能发挥了重要作用,为深入探讨肝癌的病理机制奠定了基础。Objective To investigate the effect of down-regulation of Skp2 gene expression on biological characters of HepG2 cells as well as expressions of SP1 proteins and p27 proteins. Methods The lentiviral vector expressing siRNA targeting Skp2 gene was constructed, then packaged and infected to HepG2 cells. The HepG2 cells 48 h after infection were determined for cell cycle and apoptosis by flow cytometry, for invasion ability by Transwell test, and for expressions of p27 and SP1 proteins after down- regulation of Skp2 gene expression by Western blot. Results Compared with those in normal HepG2 cells, the expression of Skp2 protein in infected cells was down-regulated significantly, while that of p27 protein was up-regulated significantly. However, the growth rate of infected cells decreased significantly, the apoptosis rate increased by nearly 2 folds, and percentage of cells at Go/Gi phase increased, while the expression level of SP1 protein decreased, and the counts of infiltrated and migrated cells decreased by about 50%（P 〈 0. 05）. Conclusion The down-regulation of Skp2 protein expression inhibited the proliferation and promoted the apoptosis of HepG2 cells, decreasing the cell invasion ability. The up-regulation and p27 protein expression and down-regulation of SP1 protein expression caused by down-regulation of Skp2 protein expression might play an important role in change of above- mentioned biological characters, which laid a foundation of further study on pathological mechanism of liver cancer.

关 键 词：肝肿瘤 SKP2基因 慢病毒载体 Sp1基因 P27基因 

分 类 号：R735.7[医药卫生—肿瘤] R73-35[医药卫生—临床医学]