蛋白激酶Cε对肝癌SK-Hep-1细胞生物学行为的影响  

Influence of protein kinase Cε on biological behavior of hepatic cancer SK-Hep-1 cells

作　　者：叶志强[1] 范瑾[2] 杨跃武[3] 刘旭辉[1] 赵坤[4] 潘卫东[4] 

机构地区：[1]中山大学附属第三医院急诊科,广东广州510630 [2]暨南大学附属第一医院妇产科,广东广州510632 [3]中山大学附属第三医院中医科,广东广州510630 [4]中山大学附属第三医院肝胆外科,广东广州510630

出　　处：《中国病理生理杂志》2014年第6期994-998,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81172783)

摘　　要：目的:使用小干扰RNA(siRNA)抑制人肝癌SK-Hep-1细胞中蛋白激酶Cε(PKCε)的表达并对抑制效果进行测定,检测沉默后SK-Hep-1细胞增殖和侵袭能力的变化。方法:培养SK-Hep-1细胞,分为PKCε-siRNA组、阴性对照(NC-siRNA)组和未行处理(control)组。采用MTT比色法观察各组细胞增殖能力的变化,采用Transwell法观察各组细胞侵袭能力的变化,Western blotting观察细胞核增殖抗原Ki67、基质金属蛋白酶9(MMP-9)等功能蛋白的表达,萤光素酶报告基因检测NF-κB信号通路的活性。结果:与对照组相比,PKCε-siRNA组PKCεmRNA和蛋白表达水平都明显降低(P<0.01),Ki67和MMP-9蛋白表达水平降低,细胞增殖能力明显降低(P<0.01),PKCε-siRNA组穿过Transwell膜的细胞少于对照组(P<0.01),且NF-κB转录活性下降(P<0.01)。结论:通过siRNA技术降低PKCε表达可抑制肝癌SK-Hep-1细胞增殖及其侵袭能力,其抑制作用可能与抑制NF-κB通路的转录活性有关。AIM：To investigate the inhibitory effect of small interfering RNA （ siRNA） on the expression of protein kinase Cε（ PKCε） in human hepatoma SK-Hep-1 cells, and the biological behaviors of the transduced cells , inclu-ding proliferation and invasion , were investigated.METHODS：The cultured SK-Hep-1 cells were divided into 3 groups, including PKCε-siRNA group , negative control （ NC）-siRNA group and control group .MTT assay was used to analyze the proliferation of the SK-Hep-1 cells in the respective groups , while invasion potency was determined by Transwell assay .The protein levels of functional biomarkers such as Ki 67 and matrix metalloproteinase 9 （ MMP-9 ） were measured by Western blotting .The Luciferase reporter gene assay was used to explore the activity of the NF-κB pathway .RESULTS：PKCεex-pression in SK-Hep-1 cells transfected with PKCε-siRNA was significantly down-regulated at both mRNA and protein levels compared with that in the normal SK-Hep-1 cells （P〈0.01）, with the decreases in the protein levels of Ki67 and MMP-9. The invasion and proliferation of SK-Hep-1 cells were obviously inhibited in PKCε-siRNA group compared with control group （P〈0.01）.Furthermore, the transcriptional activity of NF-κB was down-regulated when PKCε was effectively in-hibited by PKCε-siRNA （P〈0.01）.CONCLUSION：Down-regulation of PKCεinhibits the proliferation and invasion of hepatic carcinoma cells , which might be mediated via the NF-κB signaling pathway .

关 键 词：肝肿瘤 RNA干扰 蛋白激酶CΕ 

分 类 号：R363[医药卫生—病理学]