Expression of E-selectin in Endothelial Cells Effects of Wild Type p53 Gene.
To investigate expression of E-selectin in endothelial cells(EC)and effect of wild type p53 gene transduction on its expression, flow cytometry and RT-PCR were employed to measure the E-selectin protein and mRNA, respectively. The effects of TNFalpha stimulation and p53 gene transduction on the expression of E-selectin was studied. Flow cytometric analysis revealed that there was no E-selectin expression on the surface of resting EC, although its mRNA was detectable by RT-PCR. TNFalpha (10-1 000 ku/L)induced E-selectin expression on the surface of EC in a concentration-dependent manner. The level of E-selectin mRNA was also increased with the increasing concentrations of TNFalpha. The expression of E-selectin protein was notable after 2 h of TNFalpha stimulation and reached a peak at 4-6 h, then rapidly declined to around the basic level by 12 h. Transduction of p53 gene inhibited TNFalpha-induced EC expression of E-selectin. The E-selectin positive cell proportion was reduced from(21.31+/-1.06)% to(11.83+/-0.98)%, (n = 3, P < 0.001). The E-selectin mRNA level was also reduced correspondingly. In resting EC, neither E-selectin nor its mRNA level was affected by the transduction of p53 gene. The results indicated that p53 gene inhibited TNFalpha-induced expression of E-selectin on the surface of EC, at least partially, by reducing its mRNA level.
Lin
Yong
Y
Department of Pharmacology, Capital University of Medical Sciences, Beijing 100054, China. linyong@cdm.imicams.ac.cn
Li
Jian
J
Gao
Cun-Ji
CJ
Wang
Zhong
Z