Islam Farhadul, Gopalan Vinod, Lam Alfred K
Cancer Molecular Pathology of School of Medicine, Griffith University, Gold Coast, Australia.
Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh.
Methods Mol Biol. 2018;1756:269-279. doi: 10.1007/978-1-4939-7734-5_23.
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.
RNA干扰（RNAi）是一种正常的生理机制，其中短效应反义RNA分子调节靶基因表达。它是以序列特异性方式沉默特定目的基因的强大工具，通过设计靶向关键致病基因的RNAi，可用于针对食管腺癌中的各种分子途径。可使用不同策略开发基于RNAi的食管腺癌治疗方法，包括抑制过表达的癌基因、通过干扰细胞周期蛋白及相关基因阻断细胞分裂或通过抑制抗凋亡基因增强细胞凋亡。此外，针对多药耐药基因或化疗抗性靶点的RNAi可能提供有前景的癌症治疗选择。在此，我们以食管腺癌细胞中的原癌基因MET作为模型靶点描述RNAi技术。使用表达MET shRNA的慢病毒颗粒来沉默MET基因。然后，进行蛋白质免疫印迹分析以确认MET基因敲低。
