Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 7, 2019; 25(5): 567-583
Published online Feb 7, 2019. doi: 10.3748/wjg.v25.i5.567
Effect of adipose-derived mesenchymal stem cells on hepatocellular carcinoma: In vitro inhibition of carcinogenesis
Rim Serhal, Nagib Saliba, George Hilal, Mayssam Moussa, Ghada S Hassan, Oula El Atat, Nada Alaaeddine
Rim Serhal, Mayssam Moussa, Oula El Atat, Nada Alaaeddine, Regenerative Medicine Laboratory, Faculty of Medicine, Saint-Joseph University, Beirut 1107 2180, Lebanon
Nagib Saliba, Surgery Department, Faculty of Medicine, Saint-Joseph University and Hotel-Dieu de France, Beirut 1107 2180, Lebanon
George Hilal, Cancer and Metabolism Laboratory, Faculty of Medicine, Campus of Medical Sciences, Saint-Joseph University, Beirut 1107 2180, Lebanon
Ghada S Hassan, Nada Alaaeddine, Laboratoire d’Immunologie Cellulaire et Moléculaire, Centre Hospitalier de l’Université de Montréal, Montréal, QC H2X 0A9, Canada
Author contributions: Alaaeddine N participated in the conception and design of experiments, data analysis and interpretation, manuscript writing and final approval of manuscript; Serhal R performed the majority of experiments, design of the experiments, collection or assembly of data, data analysis and interpretation and manuscript writing; Hilal G collaborated in the conception and design of experiments; Saliba N provided study material (fat tissue); Moussa M and El Atat O participated in the collection or assembly of data and technical help; Hassan GS participated in manuscript writing and validation.
Institutional review board statement: This research was conducted with ethical approval from the Saint-Joseph University ethical committee and was carried out in accordance with the approved guidelines. Adipose tissues were obtained from healthy donors undergoing an elective liposuction procedure from abdominal, hip or thigh regions after written consent in the Department of Plastic Surgery, Hotel Dieu De France Hospital, Beirut, Lebanon.
Conflict-of-interest statement: The authors have declared no conflicts of interest.
Data sharing statement: All datasets generated and analyzed in this study are available upon request from the corresponding author.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This 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 Non Commercial (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: Nada Alaaeddine, PhD, Professor, Laboratoire d’Immunologie Cellulaire et Moléculaire, Centre Hospitalier de l’Université de Montréal, Montréal, QC H2X 0A9, Canada. naladdin@gu.edu.lb
Telephone: +961-1-426926 Fax: +961-1-427927
Received: September 10, 2018
Peer-review started: September 10, 2018
First decision: October 11, 2018
Revised: December 2, 2018
Accepted: December 6, 2018
Article in press: December 6, 2018
Published online: February 7, 2019
Processing time: 143 Days and 20.9 Hours
Abstract
AIM

To investigate the effect of adipose-derived mesenchymal stem cells (ADMSCs) and their conditioned media (CM) on hepatocellular carcinoma (HCC) cell tumorigenesis.

METHODS

The proliferation rate of HepG2 and PLC-PRF-5 HCC cancer cells was measured using the trypan blue exclusion method and confirmed using the cell-counting kit 8 (commonly known as CCK-8) assay. Apoptosis was detected by flow cytometry using annexin V-FITC. Protein and mRNA expression was quantified by ELISA and real time PCR, respectively. Migration and invasion rates were performed by Transwell migration and invasion assays. Wound healing was examined to confirm the data obtained from the migration assays.

RESULTS

Our data demonstrated that when co-culturing HCC cell lines with ADMSCs or treating them with ADMSC CM, the HCC cell proliferation rate was significantly inhibited and the apoptosis rate increased. The decreased proliferation rate was accompanied by an upregulation of P53 and Retinoblastoma mRNA and a downregulation of c-Myc and hTERT mRNA levels. More notably, ADMSCs and their CM suppressed the expression of the two important markers of HCC carcinogenicity, alpha-fetoprotein and Des-gamma-carboxyprothrombin. In addition, the migration and invasion levels of HepG2 and PLC-PRF-5 cells significantly decreased, potentially through increased expression of the tissue inhibitor metalloproteinases TIMP-1, TIMP-2 and TIMP-3.

CONCLUSION

These findings shed new light on a protective and therapeutic role for ADMSCs and their CM in controlling HCC invasiveness and carcinogenesis.

Keywords: Hepatocellular carcinoma; Adipose-derived mesenchymal stem cells; Adipose-derived mesenchymal stem cell conditioned media; Proliferation; Apoptosis; Migration; Invasion

Core tip: In this study, we report the in vitro effect of adipose derived mesenchymal stem cells (ADMSCs) on HepG2 and PLC-PRF-5 liver cell lines. It is the first study to demonstrate that ADMSCs and their respective conditioned media inhibited the expression of hepatocellular carcinoma markers alpha-fetoprotein and Des-gamma-carboxy-prothrombin and decreased cancer cell invasiveness by increasing the mRNA expression of tissue inhibitor metalloproteinases TIMP-1, TIMP-2 and TIMP-3. In addition, ADMSCs significantly reduced the proliferation rate, the invasiveness and the migration of the cancer cells while inducing their apoptosis.