吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (02): 307-310.doi: 10.13481/j.1671-587x.20150220

• 基础研究 • 上一篇    下一篇

RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响

张成才1, 宋艳艳2, 周梦娇1, 崔志倩1, 张西臣3, 邢沈阳1   

  1. 1. 吉林大学动物科学学院动物科学系, 吉林 长春 130062;
    2. 吉林大学第二医院肾病内科, 吉林 长春 130041;
    3. 吉林大学动物医学学院预防兽医学系, 吉林 长春 130062
  • 收稿日期:2014-11-06 出版日期:2015-03-28 发布日期:2015-04-04
  • 通讯作者: 邢沈阳, 教授, 硕士研究生导师(Tel:0431-87835138, E-mail:xingsy6177@163.com) E-mail:xingsy6177@163.com
  • 作者简介:张成才(1989-), 男, 山东省潍坊市人, 在读动物科学硕士, 主要从事肿瘤遗传学方面的研究。
  • 基金资助:

    吉林省科技厅科技发展计划项目资助课题(2003055025,20106044);吉林省自然科学基金资助课题(20101585)

Effect of RNA interference of YWHAZ gene on proliferation of colon cancer HCT-8 cells

ZHANG Chengcai1, SONG Yanyan2, ZHOU Mengjiao1, CUI Zhiqian1, ZHANG Xichen3, XING Shenyang1   

  1. 1. Department of Animal Science, School of Animal Science, Jilin University, Changchun 130062, China;
    2. Department of Nephrology, Second Hospital, Jilin University, Changchun 130041, China;
    3. Department of Preventive Veterinary Medicine, School of Veterinary Medicine, Jilin University, Changchun 130062, China
  • Received:2014-11-06 Online:2015-03-28 Published:2015-04-04

摘要:

目的:探讨中RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响,阐明其作用机制。方法:构建靶向基因YWHAZ的siRNA载体重组质粒pGPU6/GFP/Neo-YWHAZ-792、pGPU6/GFP/Neo-YWHAZ-733、pGPU6/GFP/Neo-YWHAZ-612和pGPU6/GFP/Neo-YWHAZ-505作为RNA干扰质粒组,同时设立阴性对照质粒组(pGPU6/GFP/Neo-shNC质粒);采用荧光定量PCR检测YWHAZ mRNA的表达水平和表达抑制率;采用MTT法检测HCT-8细胞的增殖率。结果:倒置荧光显微镜下观察,细胞转染率为60%。荧光定量PCR检测,RNA干扰组YWHAZ mRNA表达水平低于阴性对照组(P<0.01),平均抑制率为52.86%。MTT检测,与阴性对照组比较,RNA干扰质粒组细胞增殖率明显降低(P<0.01),平均细胞增殖率为68.75%。结论:干扰YWHAZ可抑制该基因的转录,并抑制HCT-8细胞的增殖。

关键词: YWHAZ基因, 结肠肿瘤, HCT-8细胞, RNA干扰

Abstract:

Objective To investigate the effect of RNA interference of YWHAZ gene on the cell proliferation of colon cancer HCT-8 cells,and to elucidate the mechanism.Methods Four siRNA plasmids targeting human YWHAZ gene,including pGPU6/GFP/Neo-YWHAZ-792,pGPU6/GFP/Neo-YWHAZ-733,pGPU6/GFP/Neo-YWHAZ-612,and pGPU6/GFP/Neo-YWHAZ-505, were constructed and regarded as RNA interference plasmid groups. pGPU6/GFP/Neo-shNC plasmid was regarded as negative control plasmid group. RT-PCR was used to detect the expression of YWHAZ mRNA and the inhibitory rates of HCT-8 cells; the proliferation rate of HCT-8 cells was detected by MTT method.Results The transfection rate of cells was 60% under inverted fluorescence microscope;the RT-PCR results showed that the levels of YWHAZ mRNA in RNA interference plasmid groups were lower than those in negative control plasmid group(P<0.01),and the average inhibitory rate was 52.86%. The MTT results showed that compared with negative control plasmid group,the proliferation rates of HCT-8 cells in RNA interference plasmid groups were decreased (P<0.01),and the average proliferation rate was 68.75%.Conclusion Interference of YWHAZ gene might inhibit the transcription of the gene,and further inhibit the proliferation of HCT-8 cells.

Key words: YWHAZ gene, colon neoplasms, HCT-8 cells, RNA interference

中图分类号: 

  • R735.3