Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Mar 27, 2021; 13(3): 328-342
Published online Mar 27, 2021. doi: 10.4254/wjh.v13.i3.328
Promotive action of 2-acetylaminofluorene on hepatic precancerous lesions initiated by diethylnitrosamine in rats: Molecular study
Amany Helmy Hasanin, Eman K Habib, Nesreen El Gayar, Marwa Matboli
Amany Helmy Hasanin, Nesreen El Gayar, Clinical Pharmacology Department, Faculty of Medicine, Ain Shams University, Cairo 11318, Egypt
Eman K Habib, Anatomy and Embryology Department, Faculty of Medicine, Ain Shams University, Cairo 11318, Egypt
Marwa Matboli, Medical Biochemistry and Molecular Biology Department, Faculty of Medicine, Ain Shams University, Cairo 11381, Egypt
Author contributions: Hasanin AH and El Gayar N conceived the idea and designed the animal model and statistical analysis; Habib EK performed histochemical examinations; Matboli M conducted bioinformatics analysis, biochemical and molecular assay; all authors drafted the manuscript and critically reviewed the manuscript and approved the final version of the manuscript for publication.
Institutional animal care and use committee statement: All animal procedures were carried out in accordance with the National Institute of Health guide for the care and use of laboratory animals (NIH Publication No. 85-23, revised 1996) and were approved by the Institutional Animal Ethics Committee for Ain Shams University, Faculty of Medicine (approval No. 17585).
Conflict-of-interest statement: The authors declare no competing interest.
Data sharing statement: No additional data are available.
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 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:
Corresponding author: Marwa Matboli, MD, Adjunct Professor, Associate Professor, Medical Biochemistry and Molecular Biology Department, Faculty of Medicine, Ain Shams University, Abbassia, P.O. box 11381, Cairo 11381, Egypt.
Received: November 23, 2020
Peer-review started: November 23, 2020
First decision: December 12, 2020
Revised: December 14, 2020
Accepted: March 2, 2021
Article in press: March 2, 2021
Published online: March 27, 2021

Diethylnitrosamine (DEN) induces hepatic neoplastic lesions over a prolonged period.


To investigate the promotive action of 2-acetylaminofluorene (2-AAF) when combined with DEN in order to develop a rat model for induction of precancerous lesion and investigate the molecular mechanism underlying the activity of 2-AAF.


The pre-precancerous lesions were initiated by intraperitoneal injection of DEN for three weeks consecutively, followed by one intraperitoneal injection of 2-AAF at three different doses (100, 200 and 300 mg/kg). Rats were separated into naïve, DEN, DEN + 100 mg 2-AAF, DEN + 200 mg 2-AAF, and DEN + 300 mg 2-AAF groups. Rats were sacrificed after 10 wk and 16 wk. Liver functions, level of alpha-fetoprotein, glutathione S-transferase-P and proliferating cell nuclear antigen staining of liver tissues were performed. The mRNA level of RAB11A, BAX, p53, and Cyclin E and epigenetic regulation by long-noncoding RNA (lncRNA) RP11-513I15.6, miR-1262 (microRNA), and miR-1298 were assessed in the sera and liver tissues of the rats.


2-AAF administration significantly increased the percent area of the precancerous foci and cell proliferation along with a significant decrease in RAB11A, BAX, and p53 mRNA, and the increase in Cyclin E mRNA was associated with a marked decrease in lncRNA RP11-513I15.6 expression with a significant increase in both miR-1262 and miR-1298.


2-AFF promoted hepatic precancerous lesions initiated through DEN by decreasing autophagy, apoptosis, and tumor suppression genes, along with increased cell proliferation, in a time- and dose-dependent manner. These actions were mediated under the epigenetic regulation of lncRNA RP11-513I15.6/miR-1262/miR-1298.

Keywords: Acetylaminofluorene, Hepatic precancerous lesion, Diethylnitrosamine, Autophagy, Apoptosis, MicroRNA

Core Tip: 2-Acetylaminofluorene epigenetically regulated the expression of long-noncoding RNA RP11-513I15.6/miRNA-1262/miR-1298 (microRNA, miRNA) resulted in decrease in RAB11A, BAX, and p53 mRNA, and the increase in Cyclin E mRNA leading to increased hepatocyte proliferation and decreased apoptosis promoting hepatocellular promoted precancerous lesion in rat models.