肝细胞生长因子对滋养细胞HLX1基因的表达及侵袭能力的影响  

Effects of the hepatocyte growth factor on expression of the homeobox gene HLX1 and invasion of trophoblast cells

作　　者：贾雪芹[1] 刘海英[2] 马玉燕[2] 高凌雪 刘媛[2] 

机构地区：[1]山东大学医学院,济南250012 [2]山东大学齐鲁医院妇产科,济南250012 [3]青岛市妇幼保健医院妇产科,山东青岛266012

出　　处：《山东大学学报（医学版）》2010年第2期58-62,66,共6页Journal of Shandong University:Health Sciences

基　　金：山东省博士基金资助课题(2007BS03052);山东省博士基金资助课题(2007B503020)

摘　　要：目的探讨肝细胞生长因子（HGF）对胎盘滋养细胞株HTR-8／SVneo细胞HLX1基因的表达及侵袭能力的影响。方法常规体外培养HTR-8／SVneo细胞，用不同浓度HGF（0、10、20、50、100ng／InL）处理细胞48h，设0浓度组为对照组；用HLX1 siRNA转染细胞24h后继续以20ng／mLHGF刺激48h，分空白对照组（MC）、siRNA＋HGF组、Mc＋HGF3个实验组；应用实时荧光定量RT—PCR和蛋白印迹法测定各组细胞中HLX1 mRNA和蚤白的表达；明胶酶谱测定上清中MMP一2的表达；Matrigel侵袭实验检测各组细胞的侵袭能力。结果④HGF处理组细胞HLX1 mRNA表达水平与对照组比较上调了90％～475％（P〈0．01），且与HGF呈剂量依赖性；20ng／mLHGF使HLX1蛋白表达水平上调（156．7±6．4）％，P〈0．01；②siRNA＋HGF组与Mc组比较，细胞HLX1 mRNA和蛋白的表达水平分别下降（54．57±0．31）％及（68．44±2．48）％，P〈0．01；③20ng／mLHGF处理组细胞其上清MMP．2表达水平与对照纽比较上调（394．6±2．9）％，P〈0．01；siRNA＋HGF纽与MC组比较MMP-2表达水平下降（81．5±0．6）％，P〈0．01；④20ng／mLHGF处理的HTR-8／SVneo细胞穿过Matrigel的细胞数为（71±5）个，与对照组（50±3）个比较侵袭能力明显增强（P〈0．01）；siRNA＋HGF组穿膜细胞数明显减少为（20±4）个，MC组为（43±3）个，两组比较差异有统计学意义（P〈0．01）。结论HGF可提高HTR-8／SVneo细胞HLX1基因的表达及细胞的侵袭力，且侵袭力的增加可能与HLX1表达水平升高有关，．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 FILX1 protein increased by（ 156, 7 _± 6.4）% after being stimulated by 20 ng/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 level. HGF may play an important role in the augmentation of HLX1 expression and the invasion capacities of trophoblast cells.

关 键 词：肝细胞生长因子 同源异型盒基因HLX1 RNA干扰 滋养细胞 

分 类 号：R714.1[医药卫生—妇产科学]