基础医学与临床 ›› 2016, Vol. 36 ›› Issue (8): 1092-1097.

• 研究论文 • 上一篇    下一篇

Gli2基因沉默对结肠癌细胞系SW480增殖、迁移和侵袭能力的影响

赵向东,黄恒,丁亮,郭武斌,刘翔,冷波,马新   

  1. 四川医科大学附属中医医院
  • 收稿日期:2015-08-28 修回日期:2015-12-12 出版日期:2016-08-05 发布日期:2016-07-13
  • 通讯作者: 马新 E-mail:2546988725@qq.com
  • 基金资助:
    四川省卫生厅科研课题

Effect of Gli2 gene silencing on the proliferation,migration and invasion in colon cancer cell line SW480

  • Received:2015-08-28 Revised:2015-12-12 Online:2016-08-05 Published:2016-07-13

摘要: 目的 探讨胶质瘤相关癌基因同源蛋白2(Gli2)基因对结肠癌细胞系SW480增殖、迁移和侵袭能力的影响及相关机制。方法 构建靶向Gli2基因的shRNA慢病毒载体,转染到SW480细胞中,用荧光显微镜观察绿色荧光蛋白的表达;实验设干扰组、阴性对照组和空白组。用MTT法、倍增时间与集落形成实验检测细胞的增殖能力;用Transwell小室法检测细胞的迁移、侵袭能力;利用qRT-PCR和Western blot法检测细胞间Gli2、cyclinD 和MMP-2基因和蛋白的表达。结果 SW480细胞转染慢病毒72h后可见明显的绿色荧光蛋白表达,干扰组较阴性对照组与空白组Gli2表达降低,细胞增殖减慢,集落形成能力减弱,倍增时间延长,迁移、侵袭能力减弱,cyclinD1和MMP-2表达下降(p<0.05)。结论 沉默Gli2的表达能够抑制SW480 细胞增殖、迁移、侵袭能力,机制可能与cyclinD1和MMP-2的下调有关。

关键词: Gli2, 结肠癌, 增殖, 侵袭

Abstract: bjective To explore the effect and mechanism of Gli2 on the proliferation,migration and invasion in colon cancer cell line SW480.Methods Complementary shRNA oligonucleotides targeting the Gli2 gene was designed and inserted into lentiviral vector. Then shRNA lentivirus were transfected into SW480 cells and then fluorescence photographs were taken. The gene silencing efficiency was measured by qRT-PCR and Western blot. Interfered group,negative group and blank group were set up.MTT,doubling time and colony-forming test were used to examine the proliferation of SW480.Migration and invasion ability were evaluated by Transwell chambers assay;Meanwhile,the qRT-PCR、Western blot were used to examine cyclinD1 and MMP-2 mRNA and protein expression. Results Green fluorescent protein was observed under fluorescence microscope after 72h in SW480 cells transfected with lentivirus.Compared with the negative control group and blank control group,the expression of Gli2 in the interfered group was lower,the cells grew slower,the doubling time was longer and the colony-forming ability was decreased,migration and invasion ability were weaken,meanwhile,the expression of cyclinD1 and MMP-2 was declined significantly(p<0.05). Conclusion Silencing Gli2 gene could suppress proliferation,migration and invasion ability in colon cancer cell line SW480.its mechanism may be related to the down-regulation of cyclinD1 and MMP-2.

Key words: Gli2, colon cancer, proliferation, invasion