Role of SDF-1/CXCR4 axis in the invasion and migration of human choriocarcinoma cells and its significance in preeclampsia / 中华微生物学和免疫学杂志

ABSTRACT

Objective To investigate the effects of stromal cell-derived factor 1 (SDF-1) and an CXC chemokine receptor 4 (CXCR4) antagonist (AMD3100) on the invasion and migration capabilities of the huaman choriocarcinoma cell line JAR for further elucidating the role of SDF-1/CXCR4 axis in the pathogenesis of preeclampsia.Methods JAR cells were divided into four groups SDF-1 group (treated with 50 ng/ml of SDF-1),SDF-1+AM3100 mixed group (first treated with 100 ng/ml of AMD3100 for 2 hours and then treated with 50 ng/ml of SDF-1),AMD3100 group (treated with 100 ng/ml of AMD3100) and blank control group (without any treatment).RT-PCR was performed to detect the expression of CXCR4 at mRNA level in JAR cells.Western blot assay was used to measure the expression of CXCR4 and p-AKT at protein level.MTT assay was used to analyze the effects of different concentrations of SDF-1 (10,30,50 and 100 ng/ml) on the proliferation of JAR cells at different time points (0,24,48,72 h).Transwell invasion assay and wound-healing assay were used to test the changes in invasion and migration capabilities of JAR cells after different treatments.Results (1) Results of the RT-PCR showed that the expression of CXCR4 at mRNA level in JAR cells was increased in the SDF-1 group (1.839±0.083) as compared with that in the blank control group (1.372±0.086),AMD3100 group (0.694±0.045) or SDF-1+AM3100 mixed group (0.703±0.093).Moreover,the differences between the SDF-1 group and the other three groups were statistically significant (F=30.67,P<0.05).Compared with the blank control group,the expression of CXCR4 at mRNA level in JAR cells was decreased in the AMD3100 group (P<0.01).(2) Results of the Western blot assay showed that the expression of CXCR4 and p-AKT at protein level in JAR cells were enhanced in the SDF-1 group as compared with that in the blank control group,AMD3100 group or SDF-1+AM3100 mixed group.Compared with the blank control group,the expression of CXCR4 and p-AKT at protein level in JAR cells were inhibited in the AMD3100 group.(3) Results of the MTT assay showed that SDF-1,especially at the concentration of 50 ng/ml,could enhance the proliferation of JAR cells (P<0.05) and its best effect on proliferation was seen at 48 h.(4) Results of the Transwell invasion assay showed that the number of transmembrane cells in the SDF-1 group (70.49±2.42) was more than that in the blank control group (54.36±2.26),AMD3100 group (21.68±8.31),or SDF-1+AMD3100 mixed group (28.18±4.61).The differences between the SDF-1 group and the other three groups were statistically significant (F=116.26,P<0.01).Compared with the blank control group,the number of transmembrane cells was reduced in the AMD3100 group (P<0.05).(5) Results of the wound-healing assay showed that the relative migration distance was increased in the SDF-1 group (1.162±0.034) as compared with that in the blank control group (0.823±0.101),AMD3100 group (0.160±0.047),or SDF-1+AMD3100 mixed group (0.183±0.064).The differences between the SDF-1 group and the other three groups were statistically significant (F=30.500,P<0.05).Compared with the blank control group,the relative migration distance was decreased in the AMD3100 group (P<0.01).Conclusion The invasion and migration of huaman choriocarcinoma JAR cells can be enhanced by SDF-1,but inhibited by AMD3100.This study indicates that the blocked biological axis of SDF-1/CXCR4 may play an important role in the pathogenesis of preeclampsia through inducing abnormal activation of PI3K/AKT pathway,which results in inhibited invasion and migration of trophoblast cells and placenta abnormality.