Effect of shRNA targeted against RhoA on proliferation and migration of human colonic cancer cells.
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.
Department of Emergency Internal Medicine, The First Affiliated Hospital of Harbin Medical University Heilongjiang, Harbin, China.
Man
M
Wang
Department of Emergency Internal Medicine, The First Affiliated Hospital of Harbin Medical University Heilongjiang, Harbin, China.
Xiu-Jie
XJ
Wang
Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University Heilongjiang, Harbin, China.
Bing-Rong
BR
Liu
D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D015536:Down-Regulation; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D005838:Genotype; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010641:Phenotype; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D000077022:Survivin; D014162:Transfection; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000077022:Survivin; C066737:RHOA protein, human; D020742:rhoA GTP-Binding Protein
Colonic cancer; LoVo cells; RNA interference; RhoA