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 eukar...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.展开更多
基金This study was supported by Shandong Provincial Medicine and Health Technology Development Plan(2018WS047).
文摘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.
