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衰老的大肠上皮不仅多种生理功能下降,而且对大肠癌在内的多种衰老相关的肠道疾病易感性显著增加,但结肠上皮衰老及衰老的结肠上皮对癌症易感的分子机制仍然不清楚.为此,应用双向凝胶电泳(2-DE)技术分离青年人及老年人的正常结肠上皮的总蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)对差异表达的蛋白质点进行鉴定,免疫组化和实时定量(real-timequantitative)PCR检测部分差异蛋白,在青年人和老年人结肠上皮中的表达水平.得到了分辨率较高、重复性较好的青年人和老年人结肠上皮的2-DE图谱,质谱分析鉴定了17个结肠上皮衰老相关的蛋白质,免疫组化和real-timequantitativeRT-PCR证实了部分差异蛋白质的表达水平.研究结果提示,线粒体功能受伤、抗氧化能力下降是结肠上皮衰老的重要原因,4个差异蛋白质即guaninenucleotide-bindingproteinbetasubunit-likeprotein(Rack1)、stress-70protein、40SribosomalproteinSA和chlorideintracellularchannelprotein1可能与衰老的结肠上皮对癌症易感有关.
The aging of the large intestine epithelium not only reduces the physiological functions, but also significantly increases the susceptibility to various aging-related intestinal diseases, including colorectal cancer. However, the molecular mechanism of colon epithelial senescence and aging susceptibility to cancer remains unclear For this purpose, the total protein in normal colonic epithelium of young and old people was separated by 2-DE, image analysis identified differentially expressed protein spots, and matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF -MS) were used to identify differentially expressed protein spots, and immunohistochemistry and real-time quantitative PCR were used to detect the expression levels of some differentially expressed proteins in the colonic epithelium of young and old people. The 2-DE patterns of colonic epithelium in young adults and the elderly were analyzed by mass spectrometry, and 17 proteins related to the aging of colonic epithelium were identified. Immunohistochemistry and real-time quantitative RT-PCR confirmed the expression levels of some differentially expressed proteins. Tip, mitochondrial dysfunction, decreased antioxidant capacity is an important reason for the aging of the colon epithelium, four differential proteins that guaninenucleotide- Binding protein betasubunit-likeprotein (Rack1), stress-70protein, 40SribosomalproteinSA and chlorideintracellularchannelprotein1 may be associated with cancer susceptibility to aging colon epithelium.