shRNA干扰PLCε1基因对人食管癌Eca109细胞增殖和细胞周期的影响  

Effect of shRNA interfering PLCε1 gene on proliferation and cell cycle of human esophageal carcinoma Eca109 cells

作　　者：周荣秒[1] 李宾[1] 牛朝旭 王娜[1] 黄茜[1] 霍向然[1] 李琰[1] 

机构地区：[1]河北医科大学第四医院河北省肿瘤研究所分子生物学研究室,河北石家庄050011 [2]石家庄平安医院外科,河北石家庄050021

出　　处：《基础医学与临床》2015年第2期208-212,共5页Basic and Clinical Medicine

摘　　要：目的探讨沉默PLCε1基因对食管癌Eca109细胞增殖和细胞周期的影响及其可能的机制。方法 PLCε11、PLCε12和PLCε13质粒表达载体用于沉默PLCε1,通用阴性对照质粒表达载体HK作为对照。用阳离子脂质体进行转染,筛选出干扰效果最好的质粒表达载体(PLCε12)。实验分为Eca109组、HK组和PLCε12组。MTT检测细胞的存活率。FCM检测细胞周期,RT-PCR检测细胞P16、Cyclin D1基因mRNA表达。结果 HK、PLCε12质粒表达载体转染Eca109细胞后48和72 h,PLCε12组Eca109细胞的存活率分别为80.73%和75.88%,显著低于HK组(P<0.001)。Eca109细胞转染后24 h,PLCε12组处于S期的细胞比例明显低于HK组(P<0.01),细胞周期阻滞于G0/G1期。质粒表达载体转染Eca109细胞48 h后,PLCε12组Eca109细胞P16基因mRNA表达水平明显高于HK组(P<0.01)。结论沉默PLCε1基因可能通过上调Eca109细胞P16基因的表达,阻止细胞周期从G1期向S期的过渡,抑制Eca109细胞的增殖活性。Objective To explore the impact of silencing PLCε1 gene on proliferation and cell cycle of esophageal carcinoma Eca109 cells. Methods Three plasmid expression vectors（ PLCε11,PLCε12 and PLCε13） were constructed to silence PLCε1 gene. A negative control plasmid expression vector（ HK） was constructed at the same time to serve as a control. The plasmid expression vectors were transfected into esophageal carcinoma Eca109 cells by cations liposome.The plasmid expression vector with the best interference effect（ PLCε12） was chosen. The study included Eca109 group,HK group and PLCε12 group. Cell viability of Eca109 cells was evaluated by MTT assay. The cell cycles were detected by FCM. The mRNA expression of P16 and Cyclin D1 gene was measured by RT-PCR. Results The cell viabilitys of Eca109 cells in PLCε12 group were 80. 73% and 75. 88% at 48 and 72 h after transfection,which were significantly lower than that of Eca109 cells in HK group（ P 〈0. 001）. The percentage of S phase Eca109 cells inPLCε12 group was lower than that of Eca109 cells in HK group（ P 〈0. 01）,the cell cycle of PLCε12 group Eca109 cells was arrested in G0/ G1 phase. The P16 gene mRNA expression of PLCε12 group Eca109 cells was higher than that of HK group Eca109 cells（ P 〈0. 01）. Conclusions Silencing PLCε1 gene may up-regulate P16 gene mRNA expression and then arrest the cell cycle at G0/ G1 phase and so inhibit proliferation of Eca109 cells.

关 键 词：PLCε1基因 食管癌 RNA干扰 增殖 

分 类 号：R735.1[医药卫生—肿瘤]