ABSTRACT
Objective To observe the expression of interferon induced protein (IP)-10 and the role of nuclear factor-kappaB (NF-κB) signaling pathway in rat peritoneal mesothelial cells (RPMCs) under the action of lipopolysaccharide (LPS).Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and cultured under defined in vitro conditions.The cells were exposed respectively to different concentrations of LPS (0,10,100,1 000,10 000 ng/ml) for 3 h or treated with LPS (100 ng/ml)for different time points (0,1,3,6,12,24,48 h).For observing the effect of LPS on the expression of p-p65 and p65,the RPMCs were treated with LPS (100 ng/ml) for different time points (0,15,30,60,120 min).For observing the effect of BAY11-7085 on the expression of IP-10 mRNA,the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μ mol/L) for2 h,then treated with LPS for another 3 h,respectively.Expression of IP-10 mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR).Expression of NF-κB and p-NF-κB protein was detected by Western blot.The secretion of IP-10 was determined by enzyme-linked immunosorbent assay (ELISA).Results Compared with the control group,stimulation of RPMCs with 10 ng/ml LPS resulted in a significant increase in the expression of IP-10 mRNA (P ＜0.05).1 000 ng/ml LPS has the strongest effect on IP-10 expression compared with that of 10 ng/ml and 100 ng/ml LPS.Treatment with 100 ng/ml LPS resuhed in time-dependent increase in the gene level of IP-10,with the peak at 3 h.However,after that time point,the gene level of them was gradually attenuated.Following treatment with LPS (100 ng/ml),the level of p-NF-κB began to increase at 15 min,gradually reached the peak at 1 hour,and then decreased.But the level of which at 2 h is still significant higher than that of medium control.5 μmol/L BAY11-7085 significantly decreased the up-regulation of IP-10 induced by LPS.Conclusions LPS enhanced the expression of IP-10 on RPMCs in a concentration-dependent and a time-dependent manner.LPS induced expression of IP-10 depended on the NF-κB signal transduction pathway.
ABSTRACT
Objective To investigate the expression and clinical significance of CXCR3/CXCL10in human cancer.Methods CXCR3 and CXCL10 were detected in 60 paraffinic tissues of patients with primary breast cancer,20 of mammary fibroma and 20 of mastopathy by immunohistochemistry S-P method and two stage method.Results The expression of CXCR3 (40/60,66.7％ ) and CXCL10 (45/60,75％)in breast cancer was higher than that in mastopathy [CXCR3(8/20,40％ )x2 =4.44,P =0.035 ;CXCL10( 10/20,50％ )x2 =4.36,P =0.037)].The expression of CXCR3 was related to status of axillary lymph node metastasis,clinical stage and the expression of HER-2 (x2 =4.15,P =0.042; x2 =7.74,P =0.021 ;x2 =4.27,P =0.039).The expression of CXCR3 had positive relationship to the number axillary lymph node metastasis( rs =0.375,P =0.003 ),clinical stage ( rs =0.451,P =0.000).Conclusions CXCR3 may be related to the progression and metastasis of breast cancer,and it may be used as a marker of breast cancer prognosis.