[The effect of inhibiting EOLA1 expression in ECV304 cells].
OBJECTIVE
To study the effect of inhibiting the expression of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) on proliferation of human umbilical vein endothelial cell line ECV304.


METHODS
After constructing and transfecting EGFP-EOLA1 fusion protein expressive vector into ECV304 cells, the transfected cells was cultured in M199 containing G418 for 5 weeks to screen the cell line stable expression EGFP-EOLA1 fusion protein. Oligonucleotides targeting EOLA1 at different sites were synthesized and inserted into pSinencer3.1/H1 vector. Then, the recombinant vector was transfected into the cultured ECV304 cells and the inhibiting effect to target gene EOLA1 was investigated by observing the green fluorescence in transfected cells under inverted fluorescent microscope and by Western blot assay. The proliferation of ECV304 cells was numbered when the expression of EOLA1 in ECV304 cells was inhibited by RNA interference.


RESULTS
The ECV304 cell line stably expressing EGFP-EOLA1 fusion protein was constructed and the siEOLA1 interfere vectors can knock down EOLA1 gene expression specially. When blocking the expression of EOLA1 in ECV304 cells,the proliferation of cells slowed down.


CONCLUSIONS
EOLA1 maybe has a role on the proliferation of cells.
Liang
Zi-wen
ZW
Department of Endocrinology, Southwest Hospital, Third Military Medical University, Chongqing, 400038 PR China. liangzw@mail.tmmu.com.cn
Yang
Zong-cheng
ZC
Chen
Jian
J
Luo
Xiao-feng
XF
Wang
Xing-ming
XM
D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D015536:Down-Regulation / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D010641:Phenotype; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics
C495142:EOLA1 protein, human; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins