Kylarová Dana, Vrchovecký Jan, Holinka Martin, Erdösová Bela
Department of Histology and Embryology, Faculty of Medicine, Palacký University, Olomouc, Czech Republic.
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2004 Dec;148(2):229-32. doi: 10.5507/bp.2004.046.
In last few years, numerous groups of proteins participating in the regulation of cell proliferation, differentiation and death during ontogenesis have been described. In this study we compared the occurrence of Bcl-2, p53 and myc protein families with the level of proliferative activity and apoptosis during development of duodenal epithelium. Paraffin embedded tissues of eight human embryos and foetuses aged from the 6th-18th week of IUD were used. For the detection of apoptotic cells the TUNEL method was performed, the proliferative marker PCNA and all the proteins studied were detected by means of indirect three-step immunohistochemical method. In the 6th and 8th week of intrauterine development we observed isolated TUNEL positive epithelial cells only and this was accompanied by the disperse presence of PCNA as well as by all the studied proteins: Bcl-2, Bax, Bcl-XL, c-myc, N-myc, p53, p63 and p73. In the early foetal period of duodenal development we registered changes in PCNA and TUNEL positivity in accordance with the constitution of the stem cell pool on base of villi, where more numerous Bcl-2 positive cells were also found. The separation of primitive crypts and villi was not accompanied by any differences in distribution of Bax, Bcl-XL, c-myc, N-myc, p63 and p73 proteins between those compartments: all the studied proteins showed dispersed character. P53 rapidly decreased in this period. In the 18th week of intrauterine development the balance between proliferation in crypts and apoptosis of villi epithelium was well established and no p53 positive cells were found. In the presence of Bcl-2, Bax, Bcl-XL, p63 and p73 we did not find any dramatic changes. The myc proteins were restricted within the epithelium of the Lieberkuhn crypts only.
在过去几年中，已经描述了许多在个体发育过程中参与细胞增殖、分化和死亡调节的蛋白质组。在本研究中，我们比较了十二指肠上皮发育过程中Bcl-2、p53和myc蛋白家族的出现情况与增殖活性和细胞凋亡水平。使用了8例妊娠6至18周的人类胚胎和胎儿的石蜡包埋组织。采用TUNEL法检测凋亡细胞，采用间接三步免疫组织化学法检测增殖标志物PCNA以及所有研究的蛋白质。在子宫内发育的第6周和第8周，我们仅观察到孤立的TUNEL阳性上皮细胞，同时伴有PCNA以及所有研究蛋白质（Bcl-2、Bax、Bcl-XL、c-myc、N-myc、p53、p63和p73）的分散存在。在十二指肠发育的早期胎儿期，我们根据绒毛干细胞池的构成记录了PCNA和TUNEL阳性的变化，在那里也发现了更多的Bcl-2阳性细胞。原始隐窝和绒毛的分离并没有伴随着这些区域之间Bax、Bcl-XL、c-myc、N-myc、p63和p73蛋白分布的任何差异：所有研究的蛋白质都表现出分散的特征。在此期间p53迅速减少。在子宫内发育的第18周，隐窝增殖和绒毛上皮细胞凋亡之间的平衡已经建立良好，未发现p53阳性细胞。在存在Bcl-2、Bax、Bcl-XL、p63和p73的情况下，我们没有发现任何显著变化。myc蛋白仅局限于利伯孔隐窝的上皮内。
