Romero-Gallo Judith, Sozmen Elif G, Chytil Anna, Russell William E, Whitehead Robert, Parks W Tony, Holdren Matthew S, Her Momoko F, Gautam Shiva, Magnuson Mark, Moses Harold L, Grady William M
Department of Medicine, Vanderbilt University, Nashville, TN, USA.
Oncogene. 2005 Apr 21;24(18):3028-41. doi: 10.1038/sj.onc.1208475.
The transforming growth factor beta (TGF-beta) signaling pathway, which is activated by the TGF-beta receptor complex consisting of type I and type II TGF-beta receptors (TGFBR1 and TGFBR2), regulates cell growth and death. TGF-beta and components of its signaling pathway, particularly TGFBR2, have been implicated as tumor suppressor genes and important antimitogenic factors in the gastrointestinal tract and liver. An in vivo approach to study these effects has been hindered by the embryonic lethality of Tgfbr2(-/-) mice and poor viability of the Tgfb1(-/-) mice. Consequently, we have developed a hepatocyte-specific Tgfbr2 knockout mouse, the Alb-cre Tgfbr2(flx/flx) mouse, to study the physiologically relevant effects of TGF-beta signaling on epithelial cell proliferation in vivo. After 70% hepatectomy, we observed increased proliferation and an increased liver mass : body weight ratio in the Alb-cre Tgfbr2(flx/flx) mice compared to Tgfbr2(flx/flx) mice. We also observed decreased expression and increased phosphorylation of p130 in the livers from the Alb-cre Tgfbr2(flx/flx) mice as well as increased expression of cyclin E, which is transcriptionally regulated, in part, by p130:E2F4. Consistent with these results, in a hepatocyte cell line derived from the Tgfbr2(flx/flx) mice, we found that TGF-beta increases the nuclear localization of E2F4, and presumably the transcriptional repression of the p130:E2F4 complex. Thus, we have demonstrated that TGF-beta signaling in vivo regulates the mitogenic response in the regenerating liver, affecting the liver mass : body weight ratio after partial hepatectomy, and that these mitogenic responses are accompanied by alterations in p130 expression and phosphorylation, implicating p130 as one of the proteins regulated in vivo by TGF-beta during liver regeneration.
转化生长因子β(TGF-β)信号通路由I型和II型TGF-β受体(TGFBR1和TGFBR2)组成的TGF-β受体复合物激活,调节细胞生长和死亡。TGF-β及其信号通路的成分,特别是TGFBR2,已被认为是胃肠道和肝脏中的肿瘤抑制基因和重要的抗有丝分裂因子。研究这些作用的体内方法受到Tgfbr2(-/-)小鼠胚胎致死性和Tgfb1(-/-)小鼠生存能力差的阻碍。因此,我们开发了一种肝细胞特异性Tgfbr2基因敲除小鼠,即Alb-cre Tgfbr2(flx/flx)小鼠,以研究TGF-β信号通路对体内上皮细胞增殖的生理相关影响。在70%肝切除术后,我们观察到与Tgfbr2(flx/flx)小鼠相比,Alb-cre Tgfbr2(flx/flx)小鼠的增殖增加,肝质量与体重比增加。我们还观察到Alb-cre Tgfbr2(flx/flx)小鼠肝脏中p130的表达降低和磷酸化增加,以及细胞周期蛋白E的表达增加,细胞周期蛋白E部分受p130:E2F4转录调控。与这些结果一致,在源自Tgfbr2(flx/flx)小鼠的肝细胞系中,我们发现TGF-β增加了E2F4的核定位,推测还有p130:E2F4复合物的转录抑制。因此,我们证明了体内TGF-β信号通路调节再生肝脏中的有丝分裂反应,影响部分肝切除术后的肝质量与体重比,并且这些有丝分裂反应伴随着p130表达和磷酸化的改变,这表明p130是肝脏再生过程中体内受TGF-β调节的蛋白质之一。