Hedgehog pathway activation and epithelial-to-mesenchymal transitions during myofibroblastic transformation of rat hepatic cells in culture and cirrhosis
Hedgehog pathway activation and epithelial-to-mesenchymal transitions during myofibroblastic transformation of rat hepatic cells in culture and cirrhosis
复制标题
DOI：
10.1152/ajpgi.00292.2009
复制
发表时间：
2009-12-01
影响因子：
4.5
通讯作者：
Diehl, Anna Mae
中科院分区：
文献类型：
--
作者：
Choi, Steve S.;Omenetti, Alessia;Diehl, Anna Mae
Choi SS, Omenetti A, Witek RP, Moylan CA, Syn W, Jung Y, Yang L, Sudan DL, Sicklick JK, Michelotti GA, Rojkind M, Diehl AM。培养和肝硬化大鼠肝细胞肌成纤维转化过程中的Hedgehog通路激活和上皮到间质转化。[J] .中国生物医学工程学报，2009,31(5):559 - 561。首次发表于2009年10月8日；doi: 10.1152 / ajpgi.00292.2009。肌成纤维肝星状细胞（MF-HSC）来源于静止肝星状细胞（Q-HSC）。Q-HSC表达某些上皮细胞标记物，并已报道形成类似上皮细胞的连接复合物。我们已经证明Hedgehog （Hh）信号在HSC生长中起关键作用。由于Hh配体调节上皮-间质转化（EMT），我们确定了Q-HSC是否表达EMT标记，然后评估这些标记是否随着Q-HSC向MF-HSC转变而改变，以及这一过程是否受到Hh信号的调节。从健康肝脏中分离Q-HSC，培养促进肌成纤维细胞转变。在接受和不接受环巴胺（一种Hh抑制剂）治疗的HSC中，监测上皮和间质标志物、Hh配体和靶基因的mRNA和蛋白表达的变化。在原发性人HSC和肝硬化大鼠的克隆来源HSC中重复进行了研究。Q-HSC在体外（培养）和体内（ccl4诱导的肝硬化）激活导致Hh相互作用蛋白（Hhip， Hh拮抗剂）、EMT抑制剂骨形态发生蛋白（BMP-7）和分化抑制剂（Id2）、粘附体连接成分E-cadherin和上皮角蛋白7和19的表达降低，Gli2 （Hh靶基因）和间充质标志物（包括间充质相关转录因子Lhx2和Msx2）的表达增加。肌成纤维细胞标志物-平滑肌肌动蛋白，以及胶原蛋白等基质分子。在啮齿动物和人的HSC中，环巴胺可以逆转肌成纤维细胞的转变，降低间充质基因的表达，同时增加上皮标记物。我们得出结论，Hh信号在Q-HSC向MF-HSC的转变中起关键作用。我们的研究结果表明，Q-HSC能够在上皮细胞和间充质细胞之间转换。
Choi SS, Omenetti A, Witek RP, Moylan CA, Syn W, Jung Y, Yang L, Sudan DL, Sicklick JK, Michelotti GA, Rojkind M, Diehl AM. Hedgehog pathway activation and epithelial-to-mesenchymal transitions during myofibroblastic transformation of rat hepatic cells in culture and cirrhosis. Am J Physiol Gastrointest Liver Physiol 297: G1093-G1106, 2009. First published October 8, 2009; doi: 10.1152/ajpgi.00292.2009.-Myofibroblastic hepatic stellate cells (MF-HSC) are derived from quiescent hepatic stellate cells (Q-HSC). Q-HSC express certain epithelial cell markers and have been reported to form junctional complexes similar to epithelial cells. We have shown that Hedgehog (Hh) signaling plays a key role in HSC growth. Because Hh ligands regulate epithelial-to-mesenchymal transition (EMT), we determined whether Q-HSC express EMT markers and then assessed whether these markers change as Q-HSC transition into MF-HSC and whether the process is modulated by Hh signaling. Q-HSC were isolated from healthy livers and cultured to promote myofibroblastic transition. Changes in mRNA and protein expression of epithelial and mesenchymal markers, Hh ligands, and target genes were monitored in HSC treated with and without cyclopamine (an Hh inhibitor). Studies were repeated in primary human HSC and clonally derived HSC from a cirrhotic rat. Q-HSC activation in vitro (culture) and in vivo (CCl4-induced cirrhosis) resulted in decreased expression of Hh-interacting protein (Hhip, an Hh antagonist), the EMT inhibitors bone morphogenic protein (BMP-7) and inhibitor of differentiation (Id2), the adherens junction component E-cadherin, and epithelial keratins 7 and 19 and increased expression of Gli2 (an Hh target gene) and mesenchymal markers, including the mesenchyme-associated transcription factors Lhx2 and Msx2, the myofibroblast marker alpha-smooth muscle actin, and matrix molecules such as collagen. Cyclopamine reverted myofibroblastic transition, reducing mesenchymal gene expression while increasing epithelial markers in rodent and human HSC. We conclude that Hh signaling plays a key role in transition of Q-HSC into MF-HSC. Our findings suggest that Q-HSC are capable of transitioning between epithelial and mesenchymal fates.