Abstract:
Objective : To evaluate the effects of anti-cyclin D1 intrabody AD5N on the proliferation of HeLa cervical cancer cells.
Methods : The control plasmids (pcDNA3.1) and recombinant plasmids with intrabody (pcDNA3.1-AD5N) were stably transfected into HeLa cells using lipofectamine 2000. G418 selection was performed to obtain the positive cell clones that were named HeLa/pcDNA3.1 or Hela/pcDNA3.1-AD5N. Western blot, MTT assay and fluorescence-activated cell sorting were conducted to investigate the expression of intrabody in HeLa cells and its anti-tumor effects.
Results : AD5N was expressed efficiently in HeLa/pcDNA3.1-AD5N cells. Compared with the control, the expression of AD5N in HeLa/pcDNA3.1-AD5N cells significantly inhibited proliferation, markedly arrested the cell cycle, and induced apoptosis of HeLa cells (
P<0.01). The ratio of cells in G
0-G
1 phase was increased by 10.35%, and the percentage of cells in S phase was decreased by 15.77%. The apoptosis index was increased by 6.38%.
Conclusion : The expression of anti-cyclinD1 intrabody AD5N inhibits cell proliferation, arrests the cell cycle, and induces apoptosis of HeLa cervical cancer cells
in vitro.