Esophageal Adenocarcinoma Methods and Protocols

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MET is a receptor tyrosine kinase, act as a proto-oncogene

and promotes the different stages of pathogenesis of esophageal

adenocarcinoma [ 21 – 29 ]. Therefore, sequence-specific therapeu-

tic intervention of MET with RNAi has been shown impressive

potential in killing esophageal adenocarcinoma cells [ 23 , 27 ].

In this chapter, we describe the RNAi in esophageal adenocar-

cinoma cells using MET shRNA as a model gene. Figure 1 illus-

trates the RNAi-mediated silencing of MET in esophageal

adenocarcinoma cells. In the first step, we knockdown of MET

Table 1
RNAi approaches used in esophageal adenocarcinoma

Target

genes Methods of RNAi Remarks References

RFC3 Plasmid construct

shRNA

RFC3 suppression inhibit EAC cells growth and

proliferation

[ 30 ]

STAT3 siRNA

oligonucleotides

Enhanced apoptosis of EAC cells by blocking

anti-apoptotic survivin and BCL2L1 expression

[ 15 ]

SMAD4 siRNA

oligonucleotides

Knockdown of SMAD4 prevents TGF-β1 induced

EMT and inhibited EMT-associated invasive

behavior of EAC cells

[ 19 ]

mPGES-1 siRNA

oligonucleotides

Silencing of mPGES-1 reduced viability and increased

apoptosis of EAC cells

[ 13 ]

IGFBP2 Plasmid vector

construct

Suppression of IGFBP2 sensitized EAC cells to

cisplatin

[ 12 ]

Akt1/

Gli-1

Lentiviral vector

shRNA

Significantly inhibited the viability of EAC by

inactivating ErbB2-PI3K pathway

[ 16 ]

AGR2 Plasmid construct

shRNA

Reduced the tumor growth notably both in vitro and

in vivo

[ 20 ]

Dkk1 siRNA

oligonucleotides

Dkk1 perturbation-mediated malignant

transformation of Barratt’s esophagus

[ 31 ]

Src Lentiviral vector

shRNA

Silencing of Src sensitize Src-mutant EAC cells to

lapatinib therapy

[ 32 ]

COX7A2 siRNA

oligonucleotides

Knockdown of COX7A2 with sRNA increase the

sensitivity of EAC cells to chemo-agents and

reduced cells growth

[ 33 ]

Gli-1 siRNA

oligonucleotides

Inhibition of Gli-1 in EAC cells increased radio-

sensitivity remarkably by inactivating hedgehog

pathway

[ 34 ]

EAC esophageal adenocarcinoma, RFC3 Replication factor C subunit 3, STAT3 Signal transducer and activator of
transcription 3, SMAD4 SMAD family member 4, mPGES-1 Microsomal prostaglandin E synthase-1, IGFBP2 Insulin-
like growth factor binding protein 2, Akt1 RAC-alpha serine/threonine-protein kinase, Gli-1 Gli family Zinc finger 1,
AGR2 adenocarcinoma-associated antigen 2, Dkk1 Dickkopf-related protein 1, Src Proto-oncogene tyrosine-protein
kinase Src, COX7A2 Cytochrome C oxidase subunit 7A2

RNAi Gene Silencing and Esophageal Adenocarcinoma