Targa A C, César A C G, Cury P M, Silva A E
Departamento de Biologia, Universidade Estadual Paulista, UNESP, São José do Rio Preto, SP, Brasil.
Genet Mol Res. 2007 Sep 30;6(3):554-65.
Apoptosis has an essential function in maintaining the integrity of the gastrointestinal mucosa. Its deregulation is associated with the occurrence of lesions such as in atrophic gastritis, peptic ulcers, intestinal metaplasia, and stomach tumorigenesis. Thus, the aim of the present study was to investigate the frequency of apoptotic cells (apoptotic index, AI) by using two different immunohistochemical techniques, TUNEL and anti-activated caspase-3 antibody (CPP32), in gastric dyspepsia [chronic gastritis (CG, N = 34), chronic atrophic gastritis (CAG, N = 11), gastric ulcer (GU, N = 17), and intestinal metaplasia (IM, N = 15)], normal gastric mucosae (NM, N = 8), and gastric adenocarcinoma (GC, N = 12). The relationship was investigated between the AI and Helicobacter pylori infection, diagnosed by PCR, overexpression of p53 protein determined by immunohistochemistry, and aneuploidy by fluorescence in situ hybridization, as performed by our laboratory in previous studies. No significant differences were observed in AI between the different groups, whether by the TUNEL technique (F = 1.60; p = 0.1670) or by CPP32 antibody (F = 1.70; p = 0.1420). Nonetheless, CAG and CG groups had AI statistically higher than those of normal mucosae. These two groups (CAG and CG) also showed a higher frequency of apoptosis-positive cases (TUNEL+ or CPP32+). Generally, there was no correlation between the AI detected by the TUNEL and CPP32 techniques in the groups studied, except in the GC group (r = 0.70). Moreover, there was no significant association between apoptosis and H. pylori infection, overexpression of p53 protein and aneuploidy, but the H. pylori-positive cases only of GU (p = 0.0233) and IM (p = 0.0253) groups displayed a statistically higher AI compared to H. pylori-negative NM, when the CPP32 antibody technique was used. Thus, CG and CAG have increased apoptosis, which may occur independent of an association with H. pylori infection, aneuploidy and overexpression of p53 protein.
细胞凋亡在维持胃肠道黏膜完整性方面具有重要作用。其失调与萎缩性胃炎、消化性溃疡、肠化生及胃肿瘤发生等病变的发生有关。因此，本研究的目的是使用两种不同的免疫组织化学技术，即TUNEL法和抗活化半胱天冬酶-3抗体（CPP32），来研究胃消化不良[慢性胃炎（CG，N = 34）、慢性萎缩性胃炎（CAG，N = 11）、胃溃疡（GU，N = 17）和肠化生（IM，N = 15）]、正常胃黏膜（NM，N = 8）及胃腺癌（GC，N = 12）中凋亡细胞的频率（凋亡指数，AI）。研究了通过聚合酶链反应诊断的AI与幽门螺杆菌感染、通过免疫组织化学测定的p53蛋白过表达以及通过荧光原位杂交检测的非整倍体之间的关系，这些检测均由我们实验室在先前研究中进行。无论是通过TUNEL技术（F = 1.60；p = 0.1670）还是通过CPP32抗体（F = 1.70；p = 0.1420），不同组之间的AI均未观察到显著差异。尽管如此，CAG组和CG组的AI在统计学上高于正常黏膜组。这两组（CAG和CG）也显示出凋亡阳性病例（TUNEL+或CPP32+）的频率更高。一般来说，在所研究的组中，除了GC组（r = 0.70）外，TUNEL法和CPP32法检测到的AI之间没有相关性。此外,细胞凋亡与幽门螺杆菌感染、p53蛋白过表达及非整倍体之间没有显著关联，但当使用CPP32抗体技术时，仅GU组（p = 0.0233）和IM组（p = 0.0253）的幽门螺杆菌阳性病例与幽门螺杆菌阴性的NM相比，AI在统计学上更高。因此，CG和CAG的细胞凋亡增加，这可能独立于与幽门螺杆菌感染、非整倍体及p53蛋白过表达的关联而发生。
