人类白细胞抗原G基因表达对滋养细胞中p38丝裂原活化蛋白激酶信号通路的影响  

Effects of human leukocyte antigen-G on p38 mitogen-activated protein kinase signaling pathway in HTR-8/SVneo cell line

作　　者：李会俭[1] 顾蔚蓉[1] 李笑天[1] 

机构地区：[1]复旦大学附属妇产科医院产科,上海200011

出　　处：《中华妇产科杂志》2011年第4期271-276,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：上海市自然科学基金（10ZR1404900）

摘　　要：目的 探讨人类白细胞抗原G（HLA-G）基因表达变化对滋养细胞侵袭力、滋养细胞中p38丝裂原活化蛋白激酶（p38MAPK）及磷酸化p38MAPK（p-p38MAPK）蛋白表达的影响.方法 对滋养细胞株HTR-8/SVneo细胞进行体外培养并分组,将细胞分为实验组[转染HLA-G小分子干扰RNA（siRNA）]、阴性对照组（转染阴性对照siRNA）和空白对照组（仅转染脂质体2000）.分别用逆转录（RT）PCR技术测定转染后细胞中HLA-G mRNA的表达;蛋白印迹法测定HLA-G蛋白的表达来验证核糖核酸干扰（RNAi）敲减效率;穿膜小室侵袭实验检测细胞的侵袭力;蛋白印迹法检测p-p38MAPK积分吸光度（IA）与p38MAPK IA的比值;检测加入抑制剂SB203580后穿膜小室的侵袭细 胞数.结果 （1）HLA-G mRNA表达：实验组为0.26±0.08,阴性对照组为0.71±0.11,空白对照组为0.79±0.07.实验组与阴性对照组比较,差异有统计学意义（P＜0.01）;阴性对照组与空白对照组比较,差异无统计学意义（P＞0.05）;实验组HLA-G mRNA表达抑制率为（69.8±6.3）%,阴性对照组HLA-G mRNA表达抑制率为（14.9±2.2）%,空白对照组为0.（2）HLA-G蛋白表达：实验组为0.20±0.15,阴性对照组为0.75±0.12,空白对照组为0.76±0.21.实验组与阴性对照组比较,差异有统计学意义（P＜0.01）;阴性对照组与空白对照组比较,差异无统计学意义（P＞0.05）;实验组HLA-G蛋白表达抑制率为（81.1±14.4）%,阴性对照组HLA-G蛋白表达抑制率为（18.0±7.7）%,空白对照组为0,实验组与阴性对照组比较,差异有统计学意义（P＜0.01）.（3）穿膜小室下室的侵袭细胞数：实验组为（57±38）个,阴性对照组为（364±79）个,空白对照组为（260±84）个.实验组与阴性对照组比较,差异有统计学意义（P＜0.01）;阴性对照组与空白对照组比较,差异无统计学意义（P＞0.05）.（4）p-p38MAPK与p38MAPK蛋白比值：实验组为0.74±0.04,阴性对照组为0.47±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

关 键 词：HLA抗原 组织相容性抗原I类 滋养层 P38丝裂原活化蛋白激酶类 RNA 小分子干扰 先兆子痫 

分 类 号：R392.1[医药卫生—免疫学]