Regulation of hepatic sulfotransferase (SULT) 1E1 expression and effects on estrogenic activity in cystic fibrosis (CF).
Regulation of hepatic sulfotransferase (SULT) 1E1 expression and effects on estrogenic activity in cystic fibrosis (CF).
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DOI：
10.1016/j.jsbmb.2009.02.009
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发表时间：
2009-03
影响因子：
4.1
通讯作者：
Runge-Morris, Melissa
中科院分区：
文献类型：
--
作者：
Falany, Charles N.;He, Dongning;Li, Li;Falany, Josie L.;Wilborn, Teresa W.;Kocarek, Thomas A.;Runge-Morris, Melissa
关键词：
囊性纤维化（CF）是白种人的一种主要遗传性疾病，每2500名新生儿中就有1人患病。肝胆病理是CF发病和死亡的主要原因，仅次于肺部疾病。在CF小鼠模型的肝细胞中，SULT 1 E1活性显著升高，通常为20-30倍。SULT 1 E1通过与磺酸盐结合，负责生理浓度下β-雌二醇（E2）的灭活。SULT 1 E1活性的增加导致CF小鼠肝脏中E2调节蛋白表达的改变。为了研究人胆管细胞中CFTR的缺乏诱导肝细胞中SULT 1 E1表达的机制，开发了膜分离的人MMNK-1胆管细胞和人HepG 2肝细胞共培养系统。囊性纤维化跨膜传导调节因子（CFTR）在胆管胆管细胞中表达，但在肝细胞中不表达，而SULT 1 E1在肝细胞中表达，但在胆管细胞中不表达。通过免疫印迹和免疫组织化学分析确定，用siRNA抑制MMNK-1细胞中的CFTR表达>90%。将对照和CFTR-siRNA-MMNK-1细胞与HepG 2细胞在Transwell膜分离系统中共培养。共培养8小时后，将HepG 2细胞从暴露于MMNK-1细胞中取出并置于新鲜培养基中。24-48小时后，在HepG 2细胞中分析SULT 1 E1和所选E2调节蛋白的表达。结果表明，在与CFTR缺陷型MMNK-1细胞共培养的HepG 2细胞中，SULT 1 E1信息和活性被选择性地诱导。E2调节的蛋白质（IGF-1，GST-P1和碳酸酐酶II）的表达也改变了E2水平的降低。因此，胆管细胞中CFTR活性的丧失通过旁分泌机制刺激肝细胞中SULT 1 E1的表达。HepG 2细胞中SULT 1 E1的表达可通过固醇介导的肝脏X受体（LXR）激活诱导，但不能通过在子宫内膜中诱导SULT 1 E1的孕激素诱导。人胆管细胞/肝细胞共培养系统中的SULT 1 E1诱导与CF小鼠中观察到的结果一致并支持该结果。肝细胞基因表达的变化影响肝脏生化，并可能促进CF肝病的发展。
Cystic fibrosis (CF) is a major genetic disease in Caucasians affecting 1 in 2500 newborns. Hepatobiliary pathology is a major cause of morbidity and mortality in CF second only to pulmonary disease. SULT1E1 activity is significantly elevated, generally 20–30-fold, in hepatocytes of mouse models of CF. SULT1E1 is responsible for the inactivation of β-estradiol (E2) at physiological concentrations via conjugation with sulfonate. The increase in SULT1E1 activity results in the alteration of E2-regulated protein expression in CF mouse liver. To investigate the mechanism by which the absence of CFTR in human cholangiocytes induces SULT1E1 expression in hepatocytes, a membrane-separated human MMNK-1 cholangiocyte and human HepG2 hepatocyte co-culture system was developed. The cystic fibrosis transmembrane conductance regulator (CFTR) is expressed in bile duct cholangiocytes but not hepatocytes, whereas SULT1E1 is expressed in hepatocytes but not cholangiocytes. CFTR expression in MMNK-1 cells was inhibited with siRNA by >90% as determined by immunoblot and immunohistochemical analysis. Control and CFTR-siRNA-MMNK-1 cells were co-cultured with HepG2 cells in a Transwell membrane-separated system. After 8 h of co-culture, HepG2 cells were removed from exposure to MMNK-1 cells and placed in fresh medium. After 24–48 h, expression of SULT1E1 and selected E2-regulated proteins was analyzed in the HepG2 cells. Results demonstrated that SULT1E1 message and activity were selectively induced in HepG2 cells co-cultured with CFTR-deficient MMNK-1 cells. The expression of E2-regulated proteins (IGF-1, GST-P1 and carbonic anhydrase II) was also altered in response to decreased E2 levels. Thus, the loss of CFTR activity in cholangiocytes stimulates the expression of SULT1E1 in hepatocytes by a paracrine mechanism. SULT1E1 expression in HepG2 cells is inducible by sterol mediated liver-X-receptor (LXR) activation although not by progestins that induce SULT1E1 in the endometrium. SULT1E1 induction in the human cholangiocyte/hepatocyte co-culture system is consistent with and supports the results observed in CF mice. The changes in hepatocyte gene expression affect liver biochemistry and may facilitate the development of CF liver disease.
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影响因子：
4.3
作者：
Alvaro, Domenico;Mancino, Maria Grazia;Gaudio, Eugenio
通讯作者：
Gaudio, Eugenio
影响因子：
3.9
作者：
FALANY, CN;VAZQUEZ, ME;ROTH, JA
通讯作者：
ROTH, JA
影响因子：
4.8
作者：
Falany, JL;Falany, CN
通讯作者：
Falany, CN
影响因子：
3.3
作者：
Colombo, Carla
通讯作者：
Colombo, Carla
影响因子：
5.6
作者：
McKarns, SC;Letterio, JJ;Kaminski, NE
通讯作者：
Kaminski, NE