The Journal of Practical Medicine ›› 2020, Vol. 36 ›› Issue (21): 2894-2899.doi: 10.3969/j.issn.1006⁃5725.2020.21.002

• Basic Research • Previous Articles     Next Articles

Construction of colon cancer cell line with lentivirus⁃mediated MiR⁃ 222 stable silence and its biological effect

ZHU Chaoyang*,LAI Dongping,ZHANG Tao,DING Mingjian,ZHUO Shaoyuan,HUANG Xiaoyan   

  1. Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,China
  • Online:2020-11-10 Published:2020-11-30
  • Contact: ZHANG Tao E⁃mail:327664246@qq.com

Abstract:

Objective To construct a colon cancer HCT⁃116 cell line with stable silencing of miR⁃222,and to explore the effect of miR⁃222 on colon cancer cell proliferation,cycle,apoptosis and migration ability.Method HCT⁃116 cells were selected and TuD RNA(Tough Decoy RNA)was used to construct TUD⁃hsa⁃miR⁃222⁃3p Inhibitor lentiviral vector,which was transfected into miR⁃222 Inhibitor(miR⁃222 I)and negative control(NC). The normal group was not infected with lentivirus cells(HCT⁃116). MTS method was used to detect cellviability and Annexin V⁃FITC was used to detect cell apoptosis;flow cytometry was used to detect cell cycle;tran⁃swell cell model was used to detect cell migration ability. Results After miR⁃222 inhibitor was transfected intoHCT⁃116 cells and the transfection was confirmed by fluorescence microscopy and RT⁃PCR. Compared with that ofNC group and normal group,cell viability of miR⁃222 I group was significantly reduced;the proportion of cells inthe S phase was significantly reduced,and the early apoptosis rate was significantly increased. The cell migrationability was obviously inhibited. Conclusion Silencing of miR⁃222 gene can reduce colon cancer cell viability andblock cycle progression,promote apoptosis and inhibit cell migration.

Key words: miR?222, lentivirus, colon cancer cells, proliferation, cycle, apoptosis, migration