Involvement of proliferative and apoptotic factors in the development of hindgut in rat fetuses with ethylenethiourea-induced anorectal malformations
Involvement of proliferative and apoptotic factors in the development of hindgut in rat fetuses with ethylenethiourea-induced anorectal malformations
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增殖和凋亡因子在亚乙基硫脲致肛门直肠畸形大鼠胎儿后肠发育中的作用
DOI：
10.1016/j.acthis.2019.151466
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发表时间：
2020-01-01
影响因子：
2.5
通讯作者：
Bai, Yuzuo
中科院分区：
文献类型：
--
作者：
Long, Caiyun;Xiao, Yunxia;Bai, Yuzuo
背景：肛肠畸形（ARMs）是一种常见的先天性末端消化道畸形，但其发病机制尚不清楚。异常细胞增殖/凋亡被认为与ARMs有关。然而，目前还没有关于细胞增殖/凋亡相关基因的研究。目的：研究MYC原癌基因和肿瘤蛋白p53两个增殖/凋亡相关基因在乙烯硫脲诱导的ARMs大鼠胎儿后肠中的时空表达模式，并探讨其潜在功能。方法：采用免疫组织化学染色、western blotting和实时定量聚合酶链反应（RT-qPCR）检测MYC和p53的表达。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）和p53染色来检测凋亡细胞和p53表达细胞的共定位。结果：发生ARMs的大鼠胎儿出现泌尿生殖隔与肛管膜融合失败。在对照组中，MYC从妊娠第14天至妊娠第16天持续表达，并分布在后肠，而p53在尿道末端和后肠中检测到较弱；ARMs组MYC表达明显降低，而p53在尿道和后肠广泛高表达。Western blotting和RT-qPCR证实ARMs中MYC表达降低，p53表达升高。TUNEL和p53共染色显示凋亡细胞和p53表达细胞之间有相当大的重叠。结论：在ARMs大鼠胚胎中，c-myc和p53的表达模式被破坏，c-myc的下调和p53的上调可能与ARMs形态发生关键时间点的ARMs发育有关。
Background: Anorectal malformations (ARMs) are common congenital malformations of the terminal digestive tract, but little is known regarding their pathogenesis. Aberrant cell proliferation/apoptosis are believed to be involved in ARMs. However, there are no studies on proliferation/apoptosis-related genes.Purpose: We aimed to investigate the spatiotemporal expression patterns of two proliferation/apoptosis-related genes (MYC proto-oncogene and tumor protein p53) and explore their potential functions in the hindguts of ethylene thiourea-induced ARMs rat fetuses.Methods: MYC and p53 expression was evaluated using immunohistochemical staining, western blotting, and quantitative real-time polymerase chain reaction (RT-qPCR). Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and p53 costaining were performed to assay the colocalization of apoptotic and p53-expressing cells.Results: Rat fetuses with ARMs displayed fusion failure of the urogenital septum and cloacal membrane. In the control group, MYC was persistently expressed from gestational day (GD)14 to GD16 and distributed throughout the hindgut, while p53 was weakly detected in the terminal segment of the urethra and hindgut; in the ARMs group, MYC expression was obviously reduced, while p53 was widely and highly expressed in the urethra and hindgut. Western blotting and RT-qPCR confirmed the decrease in MYC and increase in p53 expression in ARMs. TUNEL and p53 co-staining revealed considerable overlap between apoptotic and p53-expressing cells.Conclusion: The expression patterns of c-myc and p53 were disrupted in ARMs rat embryos, and the downregulation of c-myc and upregulation of p53 might be related to the development of ARMs at the key time points of ARMs morphogenesis.