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
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Izquierdo M (2005) Short interfering RNAs as a tool for cancer gene therapy. Cancer Gene Ther 12:217–227
Li CX, Parker A, Menocal E et al (2006) Delivery of RNA interference. Cell Cycle 5:2103–2109
Saurabh S, Vidyarthi AS, Prasad D (2014) RNA interference: concept to reality in crop improvement. Planta 239:543–564
Takeshita F, Ochiya T (2006) Therapeutic potential of RNA interference against cancer. Cancer Sci 97:689–696
Fire A, Xu S, Montgomery MK et al (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811
Hamilton AJ, Baulcombe DC (1999) A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286:950–952
Hayashi R, Schnabl J, Handler D et al (2016) Genetic and mechanistic diversity of piRNA 3′-end formation. Nature 539:588–592
Siomi MC, Sato K, Pezic D et al (2011) PIWI-interacting small RNAs: the vanguard of genome defence. Nat Rev Mol Cell Biol 12:246–258
Ahlquist P (2002) RNA-dependent RNA polymerases, viruses, and RNA silencing. Science 296:1270–1273
Liu Q, Rand TA, Kalidas S et al (2003) R2D2, a bridge between the initiation and effector steps of the Drosophila RNAi pathway. Science 301:1921–1925
Weh KM, Howell AB, Kresty LA (2016) Expression, modulation, and clinical correlates of the autophagy protein Beclin-1 in esophageal adenocarcinoma. Mol Carcinog 55:1876–1885
Myers AL, Lin L, Nancarrow DJ et al (2015) IGFBP2 modulates the chemoresistant phenotype in esophageal adenocarcinoma. Oncotarget 6:25897–25916
Beales IL, Ogunwobi OO (2010) Microsomal prostaglandin E synthase-1 inhibition blocks proliferation and enhances apoptosis in esophageal adenocarcinoma cells without affecting endothelial prostacyclin production. Int J Cancer 126:2247–2255
Hong J, Resnick M, Behar J et al (2011) Role of Rac1 in regulation of NOX5-S function in Barrett’s esophageal adenocarcinoma cells. Am J Physiol Cell Physiol 301:C413–C420
Beales IL, Ogunwobi OO (2009) Glycine-extended gastrin inhibits apoptosis in Barrett’s esophageal and esophageal adenocarcinoma cells through JAK2/STAT3 activation. J Mol Endocrinol 42:305–318
Kebenko M, Drenckhan A, Gros SJ et al (2015) ErbB2 signaling activates the Hedgehog pathway via PI3K-Akt in human esophageal adenocarcinoma: identification of novel targets for concerted therapy concepts. Cell Signal 27:373–381
MacFarlane LA, Gu Y, Casson AG et al (2010) Regulation of fibroblast growth factor-2 by an endogenous antisense RNA and by argonaute-2. Mol Endocrinol 24:800–812
Pierini R, Kroon PA, Guyot S et al (2008) The procyanidin-mediated induction of apoptosis and cell-cycle arrest in esophageal adenocarcinoma cells is not dependent on p21(Cip1/WAF1). Cancer Lett 270:234–241
Rees JR, Onwuegbusi BA, Save VE (2006) In vivo and in vitro evidence for transforming growth factor-beta1-mediated epithelial to mesenchymal transition in esophageal adenocarcinoma. Cancer Res 66:9583–9590
Wang Z, Hao Y, Lowe AW (2008) The adenocarcinoma-associated antigen, AGR2, promotes tumor growth, cell migration, and cellular transformation. Cancer Res 68:492–497
Gentile A, Trusolino L, Comoglio PM (2008) The Met tyrosine kinase receptor in development and cancer. Cancer Metastasis Rev 27:85–94
Tuynman JB, Lagarde SM, Ten Kate FJ et al (2008) Met expression is an independent prognostic risk factor in patients with esophageal adenocarcinoma. Br J Cancer 98:1102–1108
Hack SP, Bruey JM, Koeppen H (2014) HGF/MET-directed therapeutics in gastresophageal cancer: a review of clinical and biomarker development. Oncotarget 5:2866–2880
Jardim DL, de MeloGagliato D, Falchook GS et al (2014) MET aberrations and c-MET inhibitors in patients with gastric and esophageal cancers in a phase I unit. Oncotarget 5:1837–1845
Mesteri I, Schoppmann SF, Preusser M et al (2014) Overexpression of CMET is associated with signal transducer and activator of transcription 3 activation and diminished prognosis in esophageal adenocarcinoma but not in squamous cell carcinoma. Eur J Cancer 50:1354–1360
Lennerz JK, Kwak EL, Ackerman A et al (2011) MET amplification identifies a small and aggressive subgroup of esophagogastric adenocarcinoma with evidence of responsiveness to crizotinib. J Clin Oncol 29:4803–4810
Watson GA, Zhang X, Stang MT et al (2006) Inhibition of c-Met as a therapeutic strategy for esophageal adenocarcinoma. Neoplasia 8:949–955
Anderson MR, Harrison R, Atherfold PA et al (2006) Met receptor signaling: a key effector in esophageal adenocarcinoma. Clin Cancer Res 12:5936–5943
Herrera LJ, El-Hefnawy T, Queiroz de Oliveira PE (2005) The HGF receptor c-Met is overexpressed in esophageal adenocarcinoma. Neoplasia 7:75–84
Lockwood WW, Thu KL, Lin L et al (2012) Integrative genomics identified RFC3 as an amplified candidate oncogene in esophageal adenocarcinoma. Clin Cancer Res 18:1936–1946
Lyros O, Rafiee P, Nie L et al (2015) Wnt/β-Catenin signaling activation beyond robust nuclear β-Catenin accumulation in non dysplastic Barrett’s esophagus: regulation via Dickkopf-1. Neoplasia 17:598–611
Hong YS, Kim J, Pectasides E et al (2014) Src mutation induces acquired lapatinib resistance in ERBB2-amplified human gastresophageal adenocarcinoma models. PLoS One 9:e109440
Aichler M, Elsner M, Ludyga N et al (2013) Clinical response to chemotherapy in esophageal adenocarcinoma patients is linked to defects in mitochondria. J Pathol 230:410–419
Sims-Mourtada J, Izzo JG, Apisarnthanarax S et al (2006) Hedgehog: an attribute to tumor regrowth after chemoradiotherapy and a target to improve radiation response. Clin Cancer Res 12:6565–6572
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Islam, F., Gopalan, V., Lam, A.K. (2018). RNA Interference-Mediated Gene Silencing in Esophageal Adenocarcinoma. In: Lam, A. (eds) Esophageal Adenocarcinoma. Methods in Molecular Biology, vol 1756. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7734-5_23
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DOI: https://doi.org/10.1007/978-1-4939-7734-5_23
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