Poindexter Shenika V, Reddy Vishruth K, Mittal Mukul K, Williams Amanda M, Washington M Kay, Harris Elizabeth, Mah Amanda, Hiebert Scott W, Singh Kshipra, Chaturvedi Rupesh, Wilson Keith T, Lund P Kay, Williams Christopher S
Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, Tennessee; Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, Tennessee; and.
Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, Tennessee; Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, Tennessee; and Medical Scientist Training Program, Vanderbilt University School of Medicine, Nashville, Tennessee.
Am J Physiol Gastrointest Liver Physiol. 2015 Mar 15;308(6):G562-71. doi: 10.1152/ajpgi.00253.2014. Epub 2015 Jan 8.
Myeloid translocation genes (MTGs) are transcriptional corepressors implicated in development, malignancy, differentiation, and stem cell function. While MTG16 loss renders mice sensitive to chemical colitis, the role of MTG16 in the small intestine is unknown. Histological examination revealed that Mtg16(-/-) mice have increased enterocyte proliferation and goblet cell deficiency. After exposure to radiation, Mtg16(-/-) mice exhibited increased crypt viability and decreased apoptosis compared with wild-type (WT) mice. Flow cytometric and immunofluorescence analysis of intestinal epithelial cells for phospho-histone H2A.X also indicated decreased DNA damage and apoptosis in Mtg16(-/-) intestines. To determine if Mtg16 deletion affected epithelial cells in a cell-autonomous fashion, intestinal crypts were isolated from Mtg16(-/-) mice. Mtg16(-/-) and WT intestinal crypts showed similar enterosphere forming efficiencies when cultured in the presence of EGF, Noggin, and R-spondin. However, when Mtg16(-/-) crypts were cultured in the presence of Wnt3a, they demonstrated higher enterosphere forming efficiencies and delayed progression to mature enteroids. Mtg16(-/-) intestinal crypts isolated from irradiated mice exhibited increased survival compared with WT intestinal crypts. Interestingly, Mtg16 expression was reduced in a stem cell-enriched population at the time of crypt regeneration. This is consistent with MTG16 negatively regulating regeneration in vivo. Taken together, our data demonstrate that MTG16 loss promotes radioresistance and impacts intestinal stem cell function, possibly due to shifting cellular response away from DNA damage-induced apoptosis and towards DNA repair after injury.
髓系易位基因(MTGs)是参与发育、恶性肿瘤、分化和干细胞功能的转录共抑制因子。虽然MTG16缺失使小鼠对化学性结肠炎敏感,但MTG16在小肠中的作用尚不清楚。组织学检查显示,Mtg16基因敲除(-/-)小鼠的肠上皮细胞增殖增加且杯状细胞缺乏。与野生型(WT)小鼠相比,暴露于辐射后,Mtg16(-/-)小鼠的隐窝活力增加且细胞凋亡减少。对肠上皮细胞进行磷酸化组蛋白H2A.X的流式细胞术和免疫荧光分析也表明,Mtg16(-/-)肠道中的DNA损伤和细胞凋亡减少。为了确定Mtg16缺失是否以细胞自主方式影响上皮细胞,从小鼠中分离出肠隐窝。在表皮生长因子(EGF)、诺金蛋白(Noggin)和R-促调蛋白(R-spondin)存在的情况下培养时,Mtg16(-/-)和WT肠隐窝显示出相似的成肠球形成效率。然而,当在Wnt3a存在的情况下培养Mtg16(-/-)隐窝时,它们表现出更高的成肠球形成效率,并且向成熟肠类细胞的进展延迟。与WT肠隐窝相比,从受辐射小鼠中分离出的Mtg16(-/-)肠隐窝的存活率增加。有趣的是,在隐窝再生时,干细胞富集群体中的Mtg16表达降低。这与MTG16在体内负向调节再生一致。综上所述,我们的数据表明,MTG16缺失促进了放射抗性并影响肠道干细胞功能,这可能是由于细胞反应从DNA损伤诱导的细胞凋亡转向损伤后的DNA修复。