Repression of Esophageal Neoplasia and Inflammatory Signaling by Anti-miR-31 Delivery In Vivo

Taccioli, Cristian; Garofalo, Michela; Chen, Hongping; Jiang, Yubao; Tagliazucchi, Guidantonio Malagoli; Di Leva, Gianpiero; Alder, Hansjuerg; Fadda, Paolo; Middleton, Justin; Smalley, Karl J.; Selmi, Tommaso; Naidu, Srivatsava; Farber, John L.; Croce, Carlo M.; Fong, Louise Y.

Abstract

Background:
Overexpression of microRNA-31 (miR-31) is implicated in the pathogenesis of esophageal squamous cell carcinoma (ESCC), a deadly disease associated with dietary zinc deficiency. Using a rat model that recapitulates features of human ESCC, the mechanism whereby Zn regulates miR-31 expression to promote ESCC is examined.

Methods:
To inhibit in vivo esophageal miR-31 overexpression in Zn-deficient rats (n = 12–20 per group), locked nucleic acid–modified anti-miR-31 oligonucleotides were administered over five weeks. miR-31 expression was determined by northern blotting, quantitative polymerase chain reaction, and in situ hybridization. Physiological miR-31 targets were identified by microarray analysis and verified by luciferase reporter assay. Cellular proliferation, apoptosis, and expression of inflammation genes were determined by immunoblotting, caspase assays, and immunohistochemistry. The miR-31 promoter in Zn-deficient esophagus was identified by ChIP-seq using an antibody for histone mark H3K4me3. Data were analyzed with t test and analysis of variance. All statistical tests were two-sided.

Results:
In vivo, anti-miR-31 reduced miR-31 overexpression (P = .002) and suppressed the esophageal preneoplasia in Zn-deficient rats. At the same time, the miR-31 target Stk40 was derepressed, thereby inhibiting the STK40-NF-κΒ–controlled inflammatory pathway, with resultant decreased cellular proliferation and activated apoptosis (caspase 3/7 activities, fold change = 10.7, P = .005). This same connection between miR-31 overexpression and STK40/NF-κΒ expression was also documented in human ESCC cell lines. In Zn-deficient esophagus, the miR-31 promoter region and NF-κΒ binding site were activated. Zn replenishment restored the regulation of this genomic region and a normal esophageal phenotype.

Conclusions:
The data define the in vivo signaling pathway underlying interaction of Zn deficiency and miR-31 overexpression in esophageal neoplasia and provide a mechanistic rationale for miR-31 as a therapeutic target for ESCC.

Citation

Taccioli, C., Garofalo, M., Chen, H., Jiang, Y., Tagliazucchi, G. M., Di Leva, G., …Fong, L. Y. (2015). Repression of Esophageal Neoplasia and Inflammatory Signaling by Anti-miR-31 Delivery In Vivo. JNCI: Journal of the National Cancer Institute, 107(11), djv220. https://doi.org/10.1093/jnci/djv220

• Journal Article Type: Article
• Acceptance Date: Jul 20, 2015
• Online Publication Date: Aug 18, 2015
• Publication Date: 2015-11
• Deposit Date: Jun 12, 2023
• Journal: Journal of the National Cancer Institute
• Print ISSN: 0027-8874
• Electronic ISSN: 1460-2105
• Publisher: Oxford University Press
• Peer Reviewed: Peer Reviewed
• Volume: 107
• Issue: 11
• Pages: djv220
• DOI: https://doi.org/10.1093/jnci/djv220
• Keywords: Cancer Research; Oncology