1.Inhibitory effects of shRNA targeting CD44v3 on invasion behavior of human colon cancer cell SW480 in vitro induced by hyaluronan
Wenlei ZHUO ; Yan WANG ; Zhengtang CHEN
China Oncology 2000;0(06):-
Purpose:To investigate the effect of short hairpin RNA (shRNA) targeting CD44v3 on adhesion and invasion behavior of human colon cancer cell line SW480 induced by hyaluronan in vitro. Methods:RNA interference plasmid including U6 promoter and expressing shRNA of CD44v3 was designed, constructed, and transfected into SW480 cell line. CD44v3 expression was examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Adhesive and invasive ability were examined by plate adhesion model and Boyden chamber model.Results:After plasmid transfection,CD44v3 expression in SW480 cell and the number of SW480 cell induced by hyaluronan adhesiving on plate or permeating septum of Boyden chamber decreased significantly (P
2.Human lung carcinoma cell A549 express vascular endothelial grouth factor-C induced by hyaluronan
Wenlei ZHUO ; Yan WANG ; Zhengtang CHEN ; Xujun AO
Journal of Medical Postgraduates 2004;0(02):-
Objective:To investigate the expression of VEGF-C in human lung carcinoma cell A549 induced by hyaluronan(HA) in vitro. Methods:A549 cells cultured in vitro were divided into three groups: control group(C): cultured with free serum medium(FSM);HA treatment group 1(HA1) and group 2(HA2): cultured with FSM containing different concentration of HA(HA1 group: 10?g/ml,HA2 group: 20?g/ml).After 48 hours,VEGF-C expression were examined by reverse transcription-polymerase chain reaction(RT-PCR)? Western blot and immunofluorescence staining. Results:Group C: both VEGF-C mRNA and protein expression were weak positive.HA1 and HA2 groups: both VEGF-C mRNA and protein expression were strong positive and increased significantly in a HA dose-depended way(P
3.Inhibitory effects of shRNA targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma A549 cells induced by hyaluronan in vitro.
Wenlei ZHUO ; Yan WANG ; Zhengtang CHEN
Chinese Journal of Lung Cancer 2006;9(3):236-240
BACKGROUNDIt has been proved that hyaluronan was involved in adhesion and invasion behavior of varied tumor cells. Layilin is a receptor of hyaluronan found recently and has close relationship with cytoskeleton. The aim of this study is to investigate the effect of short hairpin RNA (shRNA) targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma cell line A549 induced by hyaluronan in vitro.
METHODSRNA interference plasmid that included U6 promoter and could express shRNA targeting layilin was designed, constructed, and transfected into A549 cell line. Layilin expression was examined by RT-PCR, Western blot and immunofluorescence. Adhesive and invasive ability was examined by plate adhesion model and Boyden chamber model.
RESULTSAfter plasmid transfected, layilin expression in A549 cells obviously decreased (P < 0.01), and the numbers of adhesive A549 cells on plate and A549 cells permeating septum of Boyden chamber induced by hyaluronan also significantly decreased (P < 0.01).
CONCLUSIONSThe shRNA targeting layilin can efficiently inhibit the adhesive and invasive ability of A549 cells induced by hyaluronan in vitro.