Suppressor of cytokine signaling-3 improves proliferation and migration of human trophoblast cells during pre-eclampsia / 中华围产医学杂志

ABSTRACT

Objective To investigate the expression of suppressor of cytokine signaling-3 (SOCS-3) gene in placenta,its role in the pathogenesis of pre-eclampsia and its effect on proliferation and migration of HTR-8/SVneo cells.Methods Fifteen women with severe pre-eclampsia hospitalized in the First Affiliated Hospital of Nanjing Medical University from October 2010 to March 2011 and t 5 normal pregnant women during the same time period were investigated.Cultured HTR-8/SVneo cells were transfected with SOCS-3 specific small interfering RNA (siRNA) or negative siRNA as the controls.The expression of SOCS-3 mRNA and protein in placenta and these cells was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot.Cell proliferation was detected by methyl thiazolyl tetrazolium,cell cycle by flow cytometry and migration by the Transwell test.Two independent t tests were used for statistical analysis.Results The SOCS-3 mRNA and protein levels in the severe pre-eclampsia group were lower than those in the normal group (0.25±0.03 vs 0.71±0.08 and 0.21±0.05 vs 0.75±0.12,t=15.94 and 14.29,respectively,both P＜0.05).SOCS-3 mRNA and protein levels in the transfection group at 24 hours were lower than those in the negative control group (0.39±0.02 vs 1.00±0.04 and 0.003 7±0.001 4 vs 1.514 9±0.035 7,t=27.58 and 73.35,respectively,both P＜0.05).The integral absorbance values of cell proliferation in the transfection group at 48,72 and 96 hours after transfection were 0.23 ± 0.01,0.32±0.02 and 0.37± 0.02,respectively,which were lower than those in the negative control group (0.39± 0.02,0.55 ± 0.04 and 0.86± 0.04,t=2.60,6.64 and 42.44,respectively,all P＜0.05).The cell clonal formation was lower in the transfection group compared with the negative group (116± 15 vs 312±24,t=9.96,P＜0.05).The ratios of G1/G0 and S phase cells in the transfection group were (55.75±2.21) ％ and (31.59±0.83) ％,respectively,and were significantly different from those in the negative control group [(47.88± 1.87) ％ and (37.38± 1.34) ％,t=45.43 and 20.06,respectively,P＜0.05].After 48 hours,cell migration in the transfection group was lower than that in the negative control group (93 ± 11 vs 167± 17,t=21.36,P＜O.05).Conclusion SOCS-3 expression is probably involved in the pathogenesis of pre-eclampsia by being down-regulated and therefore impeding proliferation and migration of the trophoblast.