四溴双酚A(TBBPA)是一种溴化阻燃剂,可在WistarHan大鼠中诱导子宫内膜腺癌和其他子宫肿瘤;然而,TBBPA暴露的早期分子事件或生物标志物仍未知.我们研究了TBBPA对早期暴露后大鼠子宫内生长因子受体激活(磷酸-RTK)的影响。怀孕的Wistar汉族大鼠在妊娠第6天通过幼崽断奶(PND21)通过口服管饲法暴露于TBBPA(0、0.1、25、250mg/kg/天)。幼崽在子宫内暴露,通过哺乳,每天从PND22灌胃至PND90。收集子宫角(在PND21,PND33,PND90)并固定福尔马林或冷冻进行组织学检查,免疫组织化学,phospho-RTK阵列,或蛋白质印迹分析。在PND21处,磷光体RTK,FGFR2、FGFR3、TRKC和EPHA1在分歧处置浓度下均显著增长。一些磷酸-RTK也在PND33显著过表达,包括上皮生长因子受体(EGFR),成纤维细胞生长因子受体3-4(FGFR2,FGFR3,FGFR4),胰岛素样生长因子受体1(IGF1R),INSR,AXL,MERTK,PDGFRa和b,RET,酪氨酸激酶与免疫球蛋白样和EGF样结构域1和2(TIE1;TIE2),TRKA,不同剂量处理的VEGFR2和3以及EPHA1。EGFR,RTK在女性子宫内膜癌中过度表达,在PND90时,所有治疗组仍然显著增加。Erb-B2受体酪氨酸激酶2(ERBB2)和IGF1R在PND33处过度表达,并且通过PND90保持增加,尽管ERBB2在PND90处具有统计学显著性。磷酸化RTKs,FGFR3,AXL,DTK,HGFR,TRKC,在不同剂量处理下,VEGFR1和EPHB2和4在PND90也具有统计学显著性。下游效应器,磷酸化-MAPK44/42在治疗大鼠的子宫中也增加。我们的发现表明,在早期的TBBPA暴露后,RTK会失调,它们的持续激活可能有助于在以后的大鼠中观察到TBBPA诱导的子宫肿瘤。
Tetrabromobisphenol A (TBBPA) is a brominated flame retardant that induces endometrial adenocarcinoma and other uterine tumors in Wistar Han rats; however, early molecular events or biomarkers of TBBPA exposure remain unknown. We investigated the effects of TBBPA on growth factor receptor activation (phospho-RTK) in uteri of rats following early-life exposures. Pregnant Wistar Han rats were exposed to TBBPA (0, 0.1, 25, 250 mg/kg/day) via oral gavage on gestation day 6 through weaning of pups (PND 21). Pups were exposed in utero, through lactation, and by daily gavage from PND 22 to PND 90. Uterine horns were collected (at PND 21, PND 33, PND 90) and formalin-fixed or frozen for histologic, immunohistochemical, phospho-RTK arrays, or western blot analysis. At PND 21, the phosphor-RTKs, FGFR2, FGFR3, TRKC and EPHA1 were significantly increased at different treatment concentrations. Several phospho-RTKs were also significantly overexpressed at PND 33 which included epithelial growth factor receptor (EGFR), Fibroblast Growth Factor Receptor 3-4 (FGFR2, FGFR3, FGFR4), insulin-like growth factor receptor 1 (IGF1R), INSR, AXL, MERTK, PDGFRa and b, RET, Tyrosine Kinase with Immunoglobulin Like and EGF Like Domains 1 and 2 (TIE1; TIE2), TRKA, VEGFR2 and 3, and EPHA1 at different dose treatments. EGFR, an RTK overexpressed in endometrial cancer in women, remained significantly increased for all treatment groups at PND 90. Erb-B2 Receptor Tyrosine Kinase 2 (ERBB2) and IGF1R were overexpressed at PND 33 and remained increased through PND 90, although ERBB2 was statistically significant at PND 90. The phospho-RTKs, FGFR3, AXL, DTK, HGFR, TRKC, VEGFR1 and EPHB2 and 4 were also statistically significant at PND 90 at different dose treatments. The downstream effector, phospho-MAPK44/42 was also increased in uteri of treated rats. Our findings show RTKs are dysregulated following early life TBBPA exposures and their sustained activation may contribute to TBBPA-induced uterine tumors observed in rats later in life.