PKCε-siRNA对人宫颈癌ME180细胞增殖与侵袭能力影响的实验研究  

Effect of PKCε-siRNA on Proliferating and Invading Ability in Human Cervical Cancer ME180 Cellline

作　　者：李春梅[1] 张秋实[1] 高天旸[1] 张艺[1] 刘怡潼[1] 吴良芝[1] 

机构地区：[1]广东省第二人民医院,广东广州510310

出　　处：《实用妇产科杂志》2015年第1期57-60,共4页Journal of Practical Obstetrics and Gynecology

摘　　要：目的:探讨PKCε-siRNA对人宫颈癌ME180细胞蛋白激酶Cε(PKCε)mRNA、蛋白表达及ME180细胞增殖与侵袭能力影响。方法:建立PKCε-siRNA转染组(实验组)、NC-siRNA转染组(阴性对照组)、未行处理组(对照组)。Real-time PCR法检测各组细胞PKCεmRNA表达。Western-blot法检测各组PKCε、细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶2(MMP-2)蛋白表达。采用MTT比色法观察各组细胞增殖能力,采用划痕实验观察各组细胞迁移能力,采用Transwell法观察各组细胞侵袭能力,荧光素酶报告基因检测丝裂原活化蛋白激酶(MAPK)信号通路的活性。结果:与对照组相比,实验组PKCεmRNA明显降低,表达量为对照组的18.3%,两组差异有统计学意义(P<0.05)。PKCε、Cyclin D1、MMP-2蛋白表达水平明显降低。在MTT实验中,实验组细胞72小时的A值为0.517±0.031,与对照组(0.925±0.063)相比增殖能力明显降低,两组差异有统计学意义(P<0.05)。实验组迁移和侵袭能力明显低于对照组。实验组MAPK转录活性(4.5±0.2)低于对照组(16.3±0.9),差异有统计学意义(P<0.05)。结论:通过siRNA技术降低PKCε表达,可有效干扰MAPK通路,明显抑制宫颈癌ME180细胞增殖及迁移侵袭能力。Objective： To investigate the influence of PKCε-siRNA on PKCε mRNA,protein expression,and cell proliferation and invasion ability in human cervical cancer ME180 cellline. M ethods： ME180 cells were divided into PKCε-siRNA transfection group（ experimental group）,NC-siRNA transfected group（ negative control group） and no treatment group（ blank group）. Real-time PCR assay was used to detect PKCε mRNA expression. Westernblot assay was used to detect PKCε,Cyclin D1,MMP-2 protein expression. MTT assay was used to detect cell proliferation in each group. Cell scratch assay was used to detect cell migration. Transwell assay was used to observe cell invasion ability. Luciferase reporter gene assay was used to observe activity of mitogen-activated protein kinase（ MAPK） signaling pathway. Results： Compared with the control group,PKCε mRNA expression level was significantly decreased as 18. 3% of the control group with statistically significant differences（ P〈0. 05）. Moreover,PKCε,Cyclin D1,MMP-2 protein expression levels were significantly reduced. In the MTT assay,statictically significant differences was observed as the 72 h A value of experimental group were 0. 517 ± 0. 031,with decreased proliferation,compared with the control group 0. 925 ± 0. 063. Migration and invasion of the experimental group was significantly decreased compared to the control group. MAPK transcriptional activity of the experimental group（ 4. 5 ± 0. 2） is lower than the control group（ 16. 3 ± 0. 9）,with statistically significant differences（ P〈0. 05）.Conclusions： Reducing PKCε expression by siRNA technology can effectively interfere with MAPK pathway,and inhibit cell proliferation,migration and invasion abilities of ME180 cancer cells.

关 键 词：宫颈癌 RNA干扰 蛋白激酶CΕ 丝裂原活化蛋白激酶 

分 类 号：R737.33[医药卫生—肿瘤]