Abstract:

Objective    To evaluate the effects of the hepatocyte growth factor on expression of the homeobox gene HLX1 and invasion of the HTR-8/SVneo cell line． Methods  HTR-8/SVneo cells cultured in 6-well plates were stimulated with various concentrations of HGF or transfected with siRNA to silence HLX1 mRNA expression prior to HGF stimulation. RealTime-PCR and Western blot were used to detect mRNA and protein expressions of HLX1, respectively． Cell invasion was measured by the matrigel invasion assay and expression of MMP-2 in the cells was detected by quantitative zymography． Results  ① The HLX1 mRNA level was positively related with HGF concentration. The HLX1 mRNA level increased by (90475) % in the HGF groups compared to control groups, and  the expression level of HLX1 protein increased by（156.7±6.4）% after being stimulated by 20ng/mL HGF（both P＜0.01）. ② HLX1 siRNA transfection resulted in a  decrease of(54.57±0.31)% in HLX1 mRNA level and a decrease of (68.44±2.48)% in the protein expression level（both P＜0.01）in spite of the presence of HGF stimulation. ③ HGF stimulation resulted in a significant increase of (394.6±2.9)% in MMP-2 expression in the supernatant fluid, while HLX1 the down-regulated,  a  decrease of (81.5±0.6)% in spite of the presence of HGF stimulation (both P＜0.01). ④ HTR-8/SVneo cell invasion increased with an invasion number of (71±5), compared to the control group with a number of (50±3)(P＜0.01). Inhibition of HLX1 mRNA level suppressed cell invasion with a number of (20±4), compared to the MC group with a number of (43±3) (P＜0.01). Conclusion  Invasion of HTR-8/SVneo cells with HGF stimulation correlates with the HLX1 expression leve1．　HGF may play an important role in the augmentation of HLX1 expression and the invasion capacities of trophoblast cells.

Key words: Hepatocyte growth factor; Homeobox gene HLX1; RNA interference; Trophoblast cells