Effects of Rac1 silencing on invasion and migration of Daoy cells via RNA interference

Abstract

Objective To investigate the effects of deletion of Rac1 on invasion and migration in Medulloblastoma Daoy cells. Methods The expression of Racl mRNA and protein was detected in Daoy cells by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The changes of cytoskeleton were observed in Daoy cells after transfection with Rac1shRNA, then invasion and migration were recorded respectively in Daoy cells after transfection with Rac1N17 and Rac1L61. Results Rac1 mRNA and protein were overexpressed in all Daoy cells. Deletion of Racl decreased the cross-linked actin network and pseudopodia. After single-cell scratch for 24 h, the number of migrating cells transfected with RaclshRNA or Rac1N17 was 86 ±4 and 97 ±6 respectively, while that in the control plasmid (control group) was 198 ±7, there was significant difference between RaclshRNA group or RaclN17 group and control group (P<0. 01). In the cell invasion experiments, the number of invasion cells transfected with Rac1shRNA or RaclN17 was 30 ±4 and 45 ±6, while that was 78 ±7 in control group, there was significant difference between RaclshRNA group or Rac1N17 group and control group (P <0. 01). Conclusion Deletion of Rac1 can inhibit the invasion and migration of Daoy cells. Key words: Rac1; RNA interference; Invasion; Medulloblastoma