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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (2): 58-.

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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

贾雪芹1,刘海英2,马玉燕2,高凌雪3,刘媛2   

  1. 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
  • 收稿日期:2009-10-14 出版日期:2010-02-16 发布日期:2010-02-16
  • 通讯作者: 刘海英(1969- ),女,副教授,主要从事围产感染方面的研究
  • 作者简介:贾雪芹(1981- ),女,硕士研究生,主要从事围产医学的研究
  • 基金资助:

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

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

JIA Xueqin1, LIU Haiying2, MA Yuyan2, GAO Lingxue3, LIU Yuan2   

  1. 1. School of Medicine, Shangdong University, Jinan 250012, China;
    2. Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan 250012, China;
    3. Department of Obstetrics and Gynecology, Qingdao Maternal and Child Hospital, Qingdao 266012, Shandong, China
  • Received:2009-10-14 Online:2010-02-16 Published:2010-02-16

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/mL)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/mL HGF刺激48h,分空白对照组(MC)、siRNA+HGF组、MC+HGF 3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和蛋白的表达;明胶酶谱测定上清中MMP-2的表达;Matrigel侵袭实验检测各组细胞的侵袭能力。结果  ① HGF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/mL HGF使HLX1蛋白表达水平上调(156.7±6.4)%,P<0.01;② siRNA+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;③ 20ng/mL HGF处理组细胞其上清MMP-2表达水平与对照组比较上调(394.6±2.9)%,P<0.01;siRNA+HGF组与MC组比较MMP-2表达水平下降(81.5±0.6)%,P<0.01;④ 20ng/mL HGF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。

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

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

中图分类号: 

  • R714.1
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