[Establishment of human endothelial-overexpressed lipopolysaccharide-associated factor 1 compelling expression model and its effects on the proliferation of ECV304 cells].
OBJECTIVE
To design and construct the inducible expression vector of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1), in order to establish EOLA1 compelling expression model, and to observe the effects of EOLA1 compelling expression on the proliferation of ECV304 cells.


METHODS
Inducible overexpression vector pOPRSV I-EOLA1 was constructed by amplifying the open reading fragment of EOLA1 and subcloning it into the Not I site and Xho I site of pOPRSV I vector. After sequencing, the pOPRSV I-EOLA1 recombinant vector and pCMVLac I vector were co-transfected into ECV304 cells. The cells resistant to G418 and hygromycin were screened by G418 and hygromycin, so that stable transfected cell strain was obtained. The growth curve of cells with or without isopropyl-beta-D-thiogalactoside (IPTG) induction were graphed with cell counting.


RESULTS
The inducible overexpressed EOLA1 vector was constructed successfully. The proliferation of the cells with EOLA1 compelling expression after induction of IPTG (44 +/- 17) x 10(4) was significantly higher than that without IPTG induction (27 +/- 11) x 10(4), (P < 0.01).


CONCLUSIONS
Compelling expression of EOLA1 protein can enhance the proliferation of ECV304 cell.
Chen
Yu
Y
Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, The Third Military Medical University, Chongqing 400038, P.R. China.
Liang
Zi-wen
ZW
Liu
Yue-ming
YM
Zhang
Xiao-rong
XR
Su
Yong-yue
YY
Liang
Guang-ping
GP
Chen
Jian
J
D002460:Cell Line; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000166:cytology*; D015870:Gene Expression; D006801:Humans; D008070:Lipopolysaccharides; D008565:Membrane Proteins / Q000235:genetics*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
C495142:EOLA1 protein, human; D008070:Lipopolysaccharides; D008565:Membrane Proteins