抑制DEK基因表达对结肠癌HCT116细胞增殖的影响  

Inhibition of DEK expression reduces proliferation of human colorectal adenocarcinoma cell line HCT116

作　　者：杨静[1] 宋扬[1] 赵鹏[1] 王宏[1] 闵婕[1] 张贺龙[1] 苏海川[1] 冯英明[1] 

机构地区：[1]第四军医大学唐都医院肿瘤科,陕西西安710038

出　　处：《现代肿瘤医学》2013年第4期695-698,共4页Journal of Modern Oncology

摘　　要：目的:通过siRNA技术,阻断结肠癌细胞系HCT116中DEK基因的表达,并研究DEK基因沉默后对HCT116细胞增殖能力影响。方法:用真核转录载体pLKO.1-TRC cloning vector构建针对DEK基因的重组siRNA真核转录载体pLKO.1-siDEK A,pLKO.1-siDEK B和pLKO.1-siDEK C,脂质体法转染结肠癌细胞系HCT116,Western blot检测DEK的表达变化;选择沉默效率最高的载体,采用噻唑蓝(MTT)比色法、克隆形成实验检测转染HCT116细胞在增殖、克隆形成等方面的改变,流式细胞术检测其细胞周期的改变。结果:重组载体pLKO.1-siDEK A有效地阻断了HCT116细胞中DEK基因的表达。pLKO.1-siDEK A转染HCT116细胞后,与对照组相比细胞生长、增殖速度明显减慢(P<0.05),克隆形成率明显下降;流式细胞术表明DEKsiRNA可导致HCT116细胞G0/G1期阻滞,S期细胞减少。结论:成功构建了可有效沉默DEK基因的siRNA干涉载体,证明沉默DEK基因对人结肠癌HCT116细胞增殖有负性调节作用。本研究为进一步阐明DEK基因在结直肠癌中的作用奠定了基础。Objective：To investigate the effect of siRNA - mediated DEK gene silencing on human colorectal cancer cell line HCT116. Methods： Three target gene fragments were cloned into pLKO. 1 - TRC cloning vector. Then transfected recombinant vectors into human co]orectal adenocarcinoma cells HCT116. The interference effects was detected by Western blot. The proliferation of HCTll6 ceils was detected by MTY method. Colony formation of I-ICT116 cells transfected with test plasmid was assayed by soft agar method. Cycle was assayed by flow Cytometry. Results： Three recombinant veetors：pLKO. 1 -siDEK A,pLKO. 1 -siDEK B and pLKO. 1 -siDEK C were construc- ted successfully. The results of Western blot indicated that pLKO. 1 - siDEK A could efficiently knock down the tran- scription and expression of DEK gene. After HCT116 cells were transfected with pLKO. 1 -siDEK A,the proliferation of HCT116 cells become slowly （ P 〈 0.05 ） ; Colony formation of HCT116 cells were decreased compare with control group. Flow cytometry showed that DEK knockdown resulted in cell cycle arrest. Conclusion： Downregulation of DEK results in significant inhibition of cell growth in HCTll6. This study provided a new clue to investigate the function of DEK gene and indicated the DEK may be a new target of gene therapy on colon carcinoma.

关 键 词：DEK 结直肠癌 增殖 SIRNA 

分 类 号：R733.62[医药卫生—肿瘤] R735.35[医药卫生—临床医学]