Swain Ian X, Kresak Adam M
Department of Pathology, School of Medicine, Case Western Reserve University, 2103 Cornell Road, Cleveland, OH 44106, USA.
Cancers (Basel). 2024 Apr 26;16(9):1678. doi: 10.3390/cancers16091678.
Colorectal tumorigenesis involves the development of aberrant crypt foci (ACF) or preneoplastic lesions, representing the earliest morphological lesion visible in colon cancer. The purpose of this study was to determine changes in protein expression in carcinogen-induced ACF as they mature and transform into adenomas. Protein expression profiles of azoxymethane (AOM)-induced F344 rat colon ACF and adenomas were compared at four time points, 4 (control), 8, 16, and 24 weeks post AOM administration ( = 9/group), with time points correlating with induction and transformation events. At each time point, micro-dissected ACF and/or adenoma tissues were analyzed across multiple quantitative two-dimensional (2D-DIGE) gels using a Cy-dye labeling technique and a pooled internal standard to quantify expression changes with statistical confidence. Western blot and subsequent network pathway mapping were used to confirm and elucidate differentially expressed ( ≤ 0.05) proteins, including changes in vinculin (; = 0.007), scinderin (; = 0.02), and profilin (; = 0.01), By determining protein expression changes in ACF as they mature and transform into adenomas, a "baseline" of altered regulatory proteins associated with adenocarcinoma development in this model has been elucidated. These data will enable future studies aimed at biomarker identification and understanding the molecular biology of intestinal tumorigenesis and adenocarcinoma maturation under varying intestinal conditions.
结直肠癌发生涉及异常隐窝灶（ACF）或癌前病变的发展，ACF是在结肠癌中可见的最早形态学病变。本研究的目的是确定致癌物诱导的ACF在成熟并转化为腺瘤过程中蛋白质表达的变化。在给予偶氮甲烷（AOM）后的4个时间点（4周（对照）、8周、16周和24周，每组n = 9）比较AOM诱导的F344大鼠结肠ACF和腺瘤的蛋白质表达谱，这些时间点与诱导和转化事件相关。在每个时间点，使用Cy染料标记技术和混合内标对经显微切割的ACF和/或腺瘤组织进行多个定量二维（二维差异凝胶电泳，2D-DIGE）凝胶分析，以有统计把握地定量表达变化。采用蛋白质印迹法及后续的网络通路映射来确认和阐明差异表达（P≤0.05）的蛋白质，包括纽蛋白（P = 0.007）、肌切蛋白（P = 0.02）和肌动蛋白结合蛋白（P = 0.01）的变化。通过确定ACF在成熟并转化为腺瘤过程中的蛋白质表达变化，已阐明了该模型中与腺癌发生相关的调节蛋白改变的“基线”。这些数据将有助于未来旨在鉴定生物标志物以及了解不同肠道条件下肠道肿瘤发生和腺癌成熟的分子生物学的研究。
