Regulation of amino acid transporters by adenoviral-mediated human insulin-like growth factor-1 in a mouse model of placental insufficiency in vivo and the human trophoblast line BeWo in vitro.
Regulation of amino acid transporters by adenoviral-mediated human insulin-like growth factor-1 in a mouse model of placental insufficiency in vivo and the human trophoblast line BeWo in vitro.
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DOI：
10.1016/j.placenta.2013.11.012
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发表时间：
2014-02
期刊：
影响因子：
3.8
通讯作者：
Habli, M.
中科院分区：
文献类型：
--
作者：
Jones, H.;Crombleholme, T.;Habli, M.
我们实验室之前的工作表明，胎盘中人胰岛素样生长因子-11 (hIGF-1) 的过度表达可以纠正宫内生长受限的小鼠、大鼠和兔模型中的胎儿体重缺陷，而不改变胎盘重量。这种效应的根本机制尚未阐明。为了研究胎盘内 IGF-1 过度表达对胎盘功能的影响，我们检测了胎盘功能不全 (PI) 小鼠模型和人类滋养层 BeWo 绒毛膜癌细胞系模型中氨基酸转运蛋白的表达和定位。对于体外人类研究，BeWo 绒毛膜癌细胞维持在 F12 完全培养基 + 10% FBS 中。将细胞在无血清对照培养基±Ad-IGF-1或Ad-LacZ中孵育48小时。使用的 MOI 为 10:1 和 100:1。在 BeWo 中，与对照相比，MOI 为 100:1 时，转染效率为 100%，Ad-IGF-1 显着增加了 IGF-1 分泌、增殖和侵袭，但减少了细胞凋亡。在体外，Ad-IGF-1 处理后氨基酸摄取增加，并与 SNAT1、2、LAT1 和 4F2hc 的 RNA 表达显着增加相关。仅 SNAT2 蛋白表达增加，但 LAT1 显示从核周位置重新定位到细胞质和细胞膜。对于体内研究，定时怀孕的动物在第 18 天被分为四组；假手术对照、子宫动脉分支结扎 (UABL)、UABL + Ad-hIGF-1 (108 PFU)、UABL + Ad-LacZ (108 PFU)。在妊娠第20天，通过剖腹产收获幼崽和胎盘。只有 LAT1 mRNA 表达发生变化，这表明 PI 模型中转运蛋白水平表达的降低可以通过 Ad-hIGF1 治疗得到部分纠正。在蛋白质水平上，Ad-hIGF1 后 System L 的 PI 降低，但仍保持在对照水平。在 PI 和 Ad-hIGF1 组中，System A 亚型受到差异性调节，SNAT2 表达减少，但 SNAT1 表达增加。增强的氨基酸异构体转运蛋白表达和重新定位到膜可能是有助于 Ad-hIGF-1 介导的胎盘功能不全纠正的重要机制。
Previous work in our laboratory demonstrated that over-expression of human insulin-like growth factor-11 (hIGF-1) in the placenta corrects fetal weight deficits in mouse, rat, and rabbit models of intrauterine growth restriction without changes in placental weight. The underlying mechanisms of this effect have not been elucidated. To investigate the effect of intra-placental IGF-1 over-expression on placental function we examined amino acid transporter expression and localization in both a mouse model of placental Insufficiency (PI) and a model of human trophoblast, the BeWo Choriocarcinoma cell line. For in vitro human studies, BeWo Choriocarcinoma cells were maintained in F12 complete medium + 10%FBS. Cells were incubated in serum-free control media ± Ad-IGF-1 or Ad-LacZ for 48 h. MOIs of 10:1 and 100:1 were utilized. In BeWo, transfection efficiency was 100% at an MOI of 100:1 and Ad-IGF-1 significantly increased IGF-1 secretion, proliferation and invasion but reduced apoptosis compared to controls. In vitro, amino acid uptake was increased following Ad-IGF-1 treatment and associated with significantly increased RNA expression of SNAT1, 2, LAT1 and 4F2hc. Only SNAT2 protein expression was increased but LAT1 showed relocalization from a perinuclear location to the cytoplasm and cell membrane. For in vivo studies, timed-pregnant animals were divided into four groups on day 18; sham-operated controls, uterine artery branch ligation (UABL), UABL + Ad-hIGF-1 (108 PFU), UABL + Ad-LacZ (108 PFU). At gestational day 20, pups and placentas were harvested by C-section. Only LAT1 mRNA expression changed, showing that a reduced expression of the transporter levels in the PI model could be partially rectified with Ad-hIGF1 treatment. At the protein level, System L was reduced in PI but remained at control levels following Ad-hIGF1. The System A isoforms were differentially regulated with SNAT2 expression diminished but SNAT1 increased in PI and Ad-hIGF1 groups. Enhanced amino acid isoform transporter expression and relocalization to the membrane may be an important mechanism contributing to Ad-hIGF-1 mediated correction of placental insufficiency.
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影响因子：
1.4
作者：
Fang, J.;Mao, D.;Fant, M. E.
通讯作者：
Fant, M. E.
DOI：
10.1152/ajpendo.00359.2005
发表时间：
2006-09-01
影响因子：
5.1
作者：
Jones, Helen N.;Ashworth, Cheryl J.;McArdle, Harry J.
通讯作者：
McArdle, Harry J.
影响因子：
5.6
作者：
KARL, PI
通讯作者：
KARL, PI
影响因子：
2.3
作者：
Mandl, M;Haas, J;Desoye, G
通讯作者：
Desoye, G
影响因子：
4.5
作者：
Verrey, F
通讯作者：
Verrey, F