Effects of human leukocyte antigen-G on p38 mitogen-activated protein kinase signaling pathway in HTR-8/SVneo cell line / 中华妇产科杂志

ABSTRACT

Objective To investigate the role of human leukocyte antigen-G ( HLA-G ) on the invasion and the molecular mechanism involved in this cellular progress in HTR-8/SVneo cell line. Methods There were three groups groups of transfection, negative control and blank control, which corresponding to treatment by HLA-G specific siRNA, negative siRNA and only lipofectamine 2000 using lipofection technology in HTR-8/SVneo cell line. The efficiency of down-regulated of HLA-G was detected by reverse transcription-polymerase chain reaction and western blot analysis in mRNA and protein level,respectively. Changes of p38 mitogen-activated protein kinases (p-p38MAPK)/p38MAPK protein levels and the cell invasion were respectively detected by western blot analysis and transwell test. Results ( 1 ) The mRNA levels of HLA-G transfection group, negative control group and blank control group were 0. 26 ±0. 08, 0. 71 ±0. 11, 0. 79 ±0. 07, respectively. There was significant difference between transfection group and negative control group ( P ＜ 0. 01 ), while there was no significant difference between negative control group and blank control group ( P ＞ 0. 05 ). The efficiencies of down-regulated of HLA-G were ( 69. 8 ±6. 3)%, ( 14. 9 ± 2. 2 )%, 0 in transfection group, negative control group and blank control group respectively in mRNA level. (2)In protein levels, HLA-G were 0. 20 ±0. 15, 0. 75 ±0. 12, 0. 76 ±0. 21 in transfection group, negative control group and blank control group, respectively. There was significant difference between transfection group and negative control group ( P ＜ 0. 01 ), whereas there was no significant difference between negative control group and blank control group ( P ＞ 0. 05 ). The efficiencies of down-regulated of HLA-G were (81. 1 ± 14.4)%, ( 18.0 ± 7.7)%, 0 in transfection group, negative control group and blank control group respectively. ( 3 ) The invasive number of transfection group, negative control group and blank control group were 57 ± 38,364 ± 79 and 260 ± 84, respectively, with a significant difference between transfection group and negative control group (P ＜ 0. 01 ). There was no significant difference between negative control group and blank control group ( P ＞ 0. 05 ). ( 4 ) The p-p38MAPK/p38MAPK values of the HLA-G transfection group, negative control group and blank control group were 0. 74 ±0.04, 0. 47 ± 0. 09 and 0. 36 ± 0. 21, respectively. HLA-G transfection group was significantly different compared with the other two groups( P ＜0. 01 ). (5)Without or with SB203580, the p-p38MAPK/ p38MAPK values of the HLA-G transfection group were 0. 89 ± 0. 09 and 0. 16 ± 0. 04, the values of negative control group and blank control group were 0.76 ±0.08, 0. 14 ±0.03 and 0.51 ±0.05, 0.03 ±0.01, respectively. There was significant difference between without SB203580 and with SB203580 ( P ＜ 0. 01 ). (6)Without or with SB203580, the invasive number of transfection group were 51 ± 13 and 90 ± 21 ,respectively,which was significantly different ( P ＜ 0. 01 ). The invasive number of negative control group and blank control group were 290 ± 52, 298 ± 33 and 290 ± 73, 264 ± 64, respeczively, which was no significant difference between without SB203580 and with SB203580 (P ＞ 0. 05 ). Conclusions HLA-G gene may regulate invasion of trophoblast-derived cell line HTR-8/SVneo via p38MAPK signaling pathway. The lower expression of HLA-G in trophoblast cells may lead to the occurrence of pathologic pregnancy.