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Updated: Jan 29, 2026
Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-β Signaling
Published on: August 3, 2018
Systems Biology Analyses Show Hyperactivation of Transforming Growth Factor-β and JNK Signaling Pathways in
Andrew E Blum1, Srividya Venkitachalam2, Durgadevi Ravillah2
1Division of Gastroenterology, Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine, Cleveland, Ohio; University Hospitals Cleveland Medical Center, Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine, Cleveland, Ohio; Division of Gastroenterology, Louis Stokes Veterans Affairs Medical Center, Cleveland, Ohio.
Background & Aims:
Esophageal adenocarcinoma (EAC) is resistant to standard chemoradiation treatments, and few targeted therapies are available. We used large-scale tissue profiling and pharmacogenetic analyses to identify deregulated signaling pathways in EAC tissues that might be targeted to slow tumor growth or progression.
Methods:
We collected 397 biopsy specimens from patients with EAC and nonmalignant Barrett's esophagus (BE), with or without dysplasia. We performed RNA-sequencing analyses and used systems biology approaches to identify pathways that are differentially activated in EAC vs nonmalignant dysplastic tissues; pathway activities were confirmed with immunohistochemistry and quantitative real-time polymerase chain reaction analyses of signaling components in patient tissue samples. Human EAC (FLO-1 and EsoAd1), dysplastic BE (CP-B, CP-C, CP-D), and nondysplastic BE (CP-A) cells were incubated with pharmacologic inhibitors or transfected with small interfering RNAs. We measured effects on proliferation, colony formation, migration, and/or growth of xenograft tumors in nude mice.
Results:
Comparisons of EAC vs nondysplastic BE tissues showed hyperactivation of transforming growth factor-β (TGFB) and/or Jun N-terminal kinase (JNK) signaling pathways in more than 80% of EAC samples. Immunohistochemical analyses showed increased nuclear localization of phosphorylated JUN and SMAD proteins in EAC tumor tissues compared with nonmalignant tissues. Genes regulated by the TGFB and JNK pathway were overexpressed specifically in EAC and dysplastic BE. Pharmacologic inhibition or knockdown of TGFB or JNK signaling components in EAC cells (FLO-1 or EsoAd1) significantly reduced cell proliferation, colony formation, cell migration, and/or growth of xenograft tumors in mice in a SMAD4-independent manner. Inhibition of the TGFB pathway in BE cell lines reduced the proliferation of dysplastic, but not nondysplastic, cells.
Conclusions:
In a transcriptome analysis of EAC and nondysplastic BE tissues, we found the TGFB and JNK signaling pathways to be hyperactivated in EACs and the genes regulated by these pathways to be overexpressed in EAC and dysplastic BE. Inhibiting these pathways in EAC cells reduces their proliferation, migration, and formation of xenograft tumors. Strategies to block the TGFB and JNK signaling pathways might be developed for treatment of EAC.
Insights
Targeting transforming growth factor-β (TGFB) and Jun N-terminal kinase (JNK) pathways shows promise for esophageal adenocarcinoma (EAC). Inhibiting these pathways in EAC cells reduced tumor growth and migration, offering new therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Signaling
Background:
- Esophageal adenocarcinoma (EAC) exhibits resistance to conventional treatments.
- Limited targeted therapies exist for EAC, necessitating novel therapeutic strategies.
Purpose of the Study:
- To identify deregulated signaling pathways in EAC tissues for targeted therapy.
- To investigate the potential of targeting transforming growth factor-β (TGFB) and Jun N-terminal kinase (JNK) signaling in EAC.
Main Methods:
- RNA-sequencing and systems biology approaches on 397 EAC and Barrett's esophagus (BE) biopsy specimens.
- Pathway activity validation using immunohistochemistry and qPCR.
- In vitro and in vivo experiments with pharmacologic inhibitors and small interfering RNAs in EAC and BE cell lines and xenografts.
Main Results:
- Hyperactivation of TGFB and/or JNK signaling pathways observed in over 80% of EAC samples.
- Increased nuclear localization of phosphorylated JUN and SMAD proteins in EAC tissues.
- Inhibition of TGFB/JNK signaling significantly reduced EAC cell proliferation, migration, and xenograft tumor growth in a SMAD4-independent manner.
- TGFB pathway inhibition reduced proliferation in dysplastic BE cells but not in non-dysplastic cells.
Conclusions:
- TGFB and JNK signaling pathways are hyperactivated in EAC and contribute to tumor progression.
- Targeting TGFB and JNK pathways represents a potential therapeutic strategy for EAC.
- These findings may lead to the development of novel treatments for esophageal adenocarcinoma.
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