RNA Interference-Mediated Gene Silencing in Esophageal Adenocarcinoma

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1756)

Abstract

RNA interference (RNAi) is a normal physiological mechanism in which a short effector antisense RNA molecule regulates target gene expression. It is a powerful tool to silence a particular gene of interest in a sequence-specific manner and can be used to target against various molecular pathways in esophageal adenocarcinoma by designing RNAi targeting key pathogenic genes. RNAi-based therapeutics against esophageal adenocarcinoma can be developed using different strategies including inhibition of overexpressed oncogenes, blocking cell division by interfering cyclins and related genes or enhancing apoptosis by suppressing anti-apoptotic genes. In addition, RNAi against multidrug resistance genes or chemo-resistance targets may provide promising cancer therapeutic options. Here, we describe RNAi technology using MET, a proto-oncogene in esophageal adenocarcinoma cells, as a model target. Lentiviral particles expressing MET shRNA was used to silence MET genes. Then, Western blot analysis was performed to confirm MET knockdown.

Key words

RNAi Esophageal adenocarcinoma Western blot siRNA shRNAs 

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Farhadul Islam
    • 1
    • 2
  • Vinod Gopalan
    • 1
  • Alfred K. Lam
    • 1
  1. 1.Cancer Molecular Pathology of School of MedicineGriffith UniversityGold CoastAustralia
  2. 2.Department of Biochemistry and Molecular BiologyUniversity of RajshahiRajshahiBangladesh

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