Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 21, 2021; 27(15): 1616-1629
Published online Apr 21, 2021. doi: 10.3748/wjg.v27.i15.1616
Effects of CXCL12 isoforms in a pancreatic pre-tumour cellular model: Microarray analysis
Monia Cecati, Matteo Giulietti, Alessandra Righetti, Berina Sabanovic, Francesco Piva
Monia Cecati, Matteo Giulietti, Alessandra Righetti, Berina Sabanovic, Francesco Piva, Department of Specialistic Clinical and Odontostomatological Sciences, Polytechnic University of Marche, Ancona 60126, Italy
Author contributions: Piva F designed and coordinated the study; Cecati M and Righetti A performed the experiments, acquired and analyzed data; Giulietti M and Sabanovic B analyzed and interpreted the data; Giulietti M, Sabanovic B and Cecati M wrote the manuscript; all authors approved the final version of the article.
Supported by Fondazione Cariverona, No. 2017.0570 (to Piva F).
Institutional review board statement: The authors declare that the requirement of the Institutional Review Board Approval Form or Document is not applicable, since the experiments did not involve humans.
Conflict-of-interest statement: The authors have no conflicts of interest to disclose.
Data sharing statement: Datasets available from the corresponding author at m.giulietti@univpm.it.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Matteo Giulietti, PhD, Academic Research, Department of Specialistic Clinical and Odontostomatological Sciences, Polytechnic University of Marche, Via Brecce Bianche, Ancona 60126, Italy. m.giulietti@univpm.it
Received: January 28, 2021
Peer-review started: January 28, 2021
First decision: February 24, 2021
Revised: March 5, 2021
Accepted: March 29, 2021
Article in press: March 29, 2021
Published online: April 21, 2021
Abstract
BACKGROUND

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of death among cancers, it is characterized by poor prognosis and strong chemoresistance. In the PDAC microenvironment, stromal cells release different extracellular components, including CXCL12. The CXCL12 is a chemokine promoting the communication between tumour and stromal cells. Six different splicing isoforms of CXCL12 are known (α, β, γ, δ, ε, θ) but their role in PDAC has not yet been characterized.

AIM

To investigate the specific role of α, β, and γ CXCL12 isoforms in PDAC onset.

METHODS

We used hTERT-HPNE E6/E7/KRasG12D (Human Pancreatic Nestin-Expressing) cell line as a pancreatic pre-tumour model and exposed it to the α, β, and γ CXCL12 isoforms. The altered expression profiles were assessed by microarray analyses and confirmed by Real-Time polymerase chain reaction. The functional enrichment analyses have been performed by Enrichr tool to highlight Gene Ontology enriched terms. In addition, wound healing assays have been carried out to assess the phenotypic changes, in terms of migration ability, induced by the α, β, and γ CXCL12 isoforms.

RESULTS

Microarray analysis of hTERT-HPNE cells treated with the three different CXCL12 isoforms highlighted that the expression of only a few genes was altered. Moreover, the α and β isoforms showed an alteration in expression of different genes, whereas γ isoform affected the expression of genes also common with α and β isoforms. The β isoform altered the expression of genes mainly involved in cell cycle regulation. In addition, all isoforms affected the expression of genes associated to cell migration, adhesion and cytoskeleton. In vitro cell migration assay confirmed that CXCL12 enhanced the migration ability of hTERT-HPNE cells. Among the CXCL12 splicing isoforms, the γ isoform showed higher induction of migration than α and β isoforms.

CONCLUSION

Our data suggests an involvement and different roles of CXCL12 isoforms in PDAC onset. However, more investigations are needed to confirm these preliminary observations.

Keywords: CXCL12, Splicing isoforms, Pancreatic cancer, Microarray, Migration, Wound healing assay

Core Tip: In the microenvironment of pancreatic ductal adenocarcinoma (PDAC), stromal cells release different extracellular components, including CXCL12, in order to communicate with cancer cells. Here, we investigated the specific role of α, β, and γ CXCL12 splicing isoforms in PDAC onset, by using a pre-tumour model. Microarray analysis suggested a role of CXCL12 in cell migration, and wound healing assays confirmed this hypothesis. In particular, γ isoform showed the highest promotion of migration. Our results shed light on the molecular basis of PDAC onset and progression.