Expression of human adrenomedullin gene in gastric adenocarcinoma and construction and identification of adrenomedullin overexpression vector and adrenomedullin-shRNA vector

• 作者: Chun-Xiao ji Fu-Hao Qiao Xiao-Qin Lu
• 来源期刊: 肿瘤微环境研究
• 学科: 医学
• 关键词: ADRENOMEDULLIN Adrenomedullin gene Gastric adenocarcinoma Lentiviral vector
• 年，卷（期）: 2020,（3）
• 页码范围: 71-78
• 页数: 8页
• 分类号: R73

摘要

Background:To explore the expression of adrenomedullin in gastric adenocarcinoma tissues,and to construct a eukaryotic expression vector that effectively overexpresses adrenomedullin gene and a short hairpin RNA eukaryotic expression vector that effectively inhibits adrenomedullin gene.This study lays the foundation for exploring the impact of adrenomedullin on solid tumors.Methods:A total of 60 samples of gastric adenocarcinoma tissues and adjacent tissues were collected.Immunohistochemical staining was used to verify the expression of adrenomedullin in gastric adenocarcinoma and the adjacent tissues.According to the adrenomedullin gene sequence in the National Center for Biotechnology Information and the design principle of the small interfering RNA target sequence,design and construct three specific short hairpin RNA expression vectors targeting the adrenomedullin gene mRNA and one homology using the lentiviral vector KLPO.1.The negative control vector,RT-PCR and Western blotting methods were used to detect the expression of the adrenomedullin gene,and the expression vector with the best inhibitory effect was selected.The eukaryotic expression vector pcDNA3 was used to construct an overexpression vector containing the full length of the adrenomedullin cDNA.RT-PCR and Western blotting methods were used to detect the expression of the adrenomedullin gene.Results:The PLKO.1-adrenomedullin with the best inhibitory effect and the human adrenomedullin gene overexpression vector pcDNA-adrenomedullin were successfully constructed and screened.Conclusion:Adrenomedullin is highly expressed in gastric cancer,and effectively inhibiting the expression of adrenomedullin in gastric cancer may have certain value in the treatment of gastric cancer.