Syndecan1慢病毒干涉质粒的构建及其对舌鳞癌CAL27细胞增殖的影响  

Effect of RNA interference of Syndecan1 gene on proliferation of tongue squamous carcinoma CAL27 cells

作　　者：王晓峰[1] 孔晨飞[2] 王伟[1] 张天夫[1] 

机构地区：[1]吉林大学中日联谊医院口腔科,吉林长春130033 [2]吉林大学中日联谊医院科学研究中心,吉林长春130033

出　　处：《中国实验诊断学》2015年第9期1447-1449,共3页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅自然科学基金课题(2012150780)

摘　　要：目的探讨RNA干扰Syndecan1基因对舌鳞癌CAL27细胞增殖的影响,阐明其作用机制。方法构建靶向基因Syndecan1的siRNA载体重组质粒pDSL-hpUGIP-Syndecan1-1834、pDSL-hpUGIP-Syndecan1-1588、pDSLhpUGIP-Syndecan1-544作为RNA干扰质粒组,同时设立阴性对照质粒组(pDSL-hpUGIP-control siRNA质粒);采用荧光定量PCR检测Syndecan1mRNA的表达水平和表达抑制率;采用细胞计数法检测CAL27细胞的增殖率。结果倒置荧光显微镜下观察,细胞感染率为90%。荧光定量PCR检测,RNA干扰组Syndecan1mRNA表达水平低于阴性对照组(P<0.01),平均抑制率为68.6%。细胞增殖率检测,与阴性对照组比较,RNA干扰质粒组细胞增殖率明显升高(P<0.01)。结论干扰Syndecan1可抑制该基因的转录,并促进CAL27细胞的增殖。Objective To investigate the effect of RNA interference of Syndecan1 gene on the cell proliferation of tongue squamous carcinoma CAL27 cells,and to elucidate the mechanism.Methods Three siRNA plasmids targeting human Syndecan1 gene,including pDSL-hpUGIP- Syndecan1-1834、pDSL-hpUGIP- Syndecan1-1588 and pDSL-hpU-GIP-Syndecan1-544,were constructed and regarded as RNA interference plasmid grous.pDSL-hpUGIP-control siRNA plasmid was regarded as negative control.RT-PCR was used to detect the expression of Syndecan1 mRNA and the in-hibitory rates of CAL27 cells was evaluated.Results The infection rate of cells was 90% under inverted fluorescence microscope;the RT-PCR results showed that the levels of Syndecan1 mRNA in RNA interference plasmid grous were lower than those in negative control group（P 〈0.01）,and the average inhibitory rate was 68.6%.Compared with nega-tive control,the proliferation rates of CAL27 cells in RNA interference plasmid grous were increased（P 〈0.01）.Con-clusion Interference of Syndecan1 gene might inhibit the transcription of the gene and further increase the prolofera-tion of CAL27 cells.

关 键 词：Syndecan1基因 舌鳞癌 CAL27细胞 RNA干扰 

分 类 号：R739.86[医药卫生—肿瘤]