Man Wang1, Xiu-Jie Wang1, Bing-Rong Liu2. 1. Department of Emergency Internal Medicine, The First Affiliated Hospital of Harbin Medical University Heilongjiang, Harbin, China. 2. Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University Heilongjiang, Harbin, China.
Abstract
OBJECTIVE: To study the effects of RhoA siRNA on the malignant phenotypes of human colorectal cancer cell line LoVo. METHODS: The siRNA expression vector pGPU6/GFP/Neo-shRNA-RhoA targeting the mRNA of RhoA and vector pGPU6/GFP/Neo-NC (as a control) were constructed, and then transfected into LoVo cells. The expression of Survivin was detected by real-time fluorescent quantitative polymerase chain reaction and Western blot. The malignant phenotypes of transfected LoVo cells, including invasive activities and adhesive capabilities, were analyzed. RESULTS: RhoA mRNA and protein level were decreased after the pshRNA-RhoA transfection. The cell adhesion rates significantly decreased in the cells transfected with pshRNA-RhoA. The migrating number of LoVo cells (26.5 ± 0.9) transfected with pshRNA-RhoA was also significantly decreased as compared with the control group (53.7 ± 1.4). CONCLUSIONS: The sequence specific shRNA against RhoA constructed in the study can block the expression of RhoA in LoVo cell effectively and specifically; Blocking the expression of RhoA in LoVo cells transfected with pshRNA-RhoA can reduce their invasive and adhesive capabilities.
OBJECTIVE: To study the effects of RhoA siRNA on the malignant phenotypes of humancolorectal cancer cell line LoVo. METHODS: The siRNA expression vector pGPU6/GFP/Neo-shRNA-RhoA targeting the mRNA of RhoA and vector pGPU6/GFP/Neo-NC (as a control) were constructed, and then transfected into LoVo cells. The expression of Survivin was detected by real-time fluorescent quantitative polymerase chain reaction and Western blot. The malignant phenotypes of transfected LoVo cells, including invasive activities and adhesive capabilities, were analyzed. RESULTS:RhoA mRNA and protein level were decreased after the pshRNA-RhoA transfection. The cell adhesion rates significantly decreased in the cells transfected with pshRNA-RhoA. The migrating number of LoVo cells (26.5 ± 0.9) transfected with pshRNA-RhoA was also significantly decreased as compared with the control group (53.7 ± 1.4). CONCLUSIONS: The sequence specific shRNA against RhoA constructed in the study can block the expression of RhoA in LoVo cell effectively and specifically; Blocking the expression of RhoA in LoVo cells transfected with pshRNA-RhoA can reduce their invasive and adhesive capabilities.
Authors: B Bouzahzah; C Albanese; F Ahmed; F Pixley; M P Lisanti; J D Segall; J Condeelis; D Joyce; A Minden; C J Der; A Chan; M Symons; R G Pestell Journal: Mol Med Date: 2001-12 Impact factor: 6.354