Effects of TRP53 upregulation by nutlin 3a on the transcriptome of interstitial cell of Cajal (ICC) stem cells

Interstitial cells of Cajal (ICC) are electrical pacemakers and mediators of neuromuscular neurotransmission in the gastrointestinal tract. Studies in organotypic cultures of intact gastric muscular wall tissues identified Kit(low)Cd44(+)Cd34(+)Insr(+)Igf1r(+) cells as local ICC progenitors (Lorincz et al., Gastroenterology 2008;134:1083-1093). Subsequently, conditionally immortalized Kit(low)Cd34(+) cells were isolated by immunomagnetic and fluorescent cell sorting and serial cloning from the gastric muscular wall of homozygous, 14-day-old H-2Kb-tsA58 and 7-day-old wild-type C57BL/6J mice and shown to share other phenotypic characteristics with in-situ ICC precursors (Bardsley et al., Gastroenterology 2010;139:942-952). ICC and their precursors undergo age-related depletion in mice and humans. These changes are associated with increased protein levels for transformation-related protein 53 (TRP53). This study utilized total RNA-sequencing to investigate the effects of TRP53 upregulation by nutlin 3a treatment (control: nutlin 3b) on the transcriptome of the ICC precursor line D2211B maintained in the verified absence of the SV40 tsA58 large T antigen.

卡哈尔间质细胞（Interstitial cells of Cajal，ICC）是胃肠道中的电起搏细胞，同时也是神经肌肉信号传递的介质。在完整胃壁肌层组织的器官型培养研究中，研究者将Kit(low)Cd44(+)Cd34(+)Insr(+)Igf1r(+)细胞鉴定为局部ICC祖细胞（Lorincz等人，《胃肠病学》，2008年；134:1083-1093）。后续研究通过免疫磁珠与荧光细胞分选技术及连续克隆法，从14日龄纯合H-2Kb-tsA58小鼠与7日龄野生型C57BL/6J小鼠的胃壁肌层中分离出条件永生化Kit(low)Cd34(+)细胞，且证实该细胞与原位ICC前体具有其他共同的表型特征（Bardsley等人，《胃肠病学》，2010年；139:942-952）。ICC及其前体在小鼠与人类体内均会随年龄增长而出现耗竭现象，该变化与转化相关蛋白53（transformation-related protein 53，TRP53）的蛋白水平升高相关。本研究采用全转录组RNA测序技术，探究了nutlin 3a处理诱导TRP53上调（对照组：nutlin 3b）对经验证确无SV40 tsA58大T抗原表达的ICC前体细胞系D2211B转录组的影响。