Rosário Pinheiro Danilo do, Harada Maria Lucia, Rodriguez Burbano Rommel Mario, Nascimento Borges Barbara do
Molecular Biology Laboratory, Instituto de Ciências Biológicas. Universidade Federal do Pará, Belém, PA, Brazil.
Human Cytogenetics Laboratory, Instituto de Ciências Biológicas. Universidade Federal do Pará, Belém, PA, Brazil.
Genet Mol Biol. 2018;41(2):496-501. doi: 10.1590/1678-4685-GMB-2016-0329. Epub 2018 May 14.
Gastric cancer (GC) remains one of the main causes of cancer-related death worldwide. There are two distinct histological types of GC: diffuse and intestinal. The latter is characterized by the presence of pre-neoplastic lesions. One of the most frequently altered enzymes in intestinal GC is COX-2, an important lesion marker. This work aimed to study COX-2 methylation and expression in N-methyl-N-Nitrosurea (MNU)-induced intestinal GC in six Sapajus apella animals. The partial promoter sequence of S. apella COX-2 gene was obtained and used to identify transcription factors and cis-regulatory element binding sites. The COX-2 methylation pattern was assessed using Methylation-Specific PCR (MSP), and expression was analyzed by immunohistochemistry (IHQ). A total of 20 samples were obtained. A 675 bp fragment of the S. apella COX-2 promoter region was obtained, and it was 99.2% and 68.2% similar to H. sapiens and S. boliviensis, respectively. Similar to humans, several transcription factors and cis-regulatory element binding sites were identified in the S. apella sequence. MSP revealed that all samples were methylated. However, IHQ results demonstrated positive COX-2 expression in all pre-neoplastic and tumoral samples. The results suggest that the analyzed fragment is not crucial in COX-2 regulation of GC in S. apella.
胃癌(GC)仍然是全球癌症相关死亡的主要原因之一。GC有两种不同的组织学类型:弥漫型和肠型。后者的特征是存在癌前病变。肠型GC中最常发生改变的酶之一是COX-2,它是一种重要的病变标志物。这项工作旨在研究六只僧帽猴(Sapajus apella)中N-甲基-N-亚硝基脲(MNU)诱导的肠型GC中COX-2的甲基化和表达情况。获得了僧帽猴COX-2基因的部分启动子序列,并用于鉴定转录因子和顺式调节元件结合位点。使用甲基化特异性PCR(MSP)评估COX-2甲基化模式,并通过免疫组织化学(IHQ)分析表达情况。共获得20个样本。获得了一段675 bp的僧帽猴COX-2启动子区域片段,它与人类和玻利维亚僧帽猴的相似性分别为99.2%和68.2%。与人类相似,在僧帽猴序列中鉴定出了几种转录因子和顺式调节元件结合位点。MSP显示所有样本均发生甲基化。然而,IHQ结果表明,所有癌前和肿瘤样本中COX-2均呈阳性表达。结果表明,所分析的片段在僧帽猴GC的COX-2调节中并非关键因素。