Observational Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Dec 27, 2021; 13(12): 2137-2149
Published online Dec 27, 2021. doi: 10.4254/wjh.v13.i12.2137
New stem cell autophagy surrogate diagnostic biomarkers in early-stage hepatocellular carcinoma in Egypt: A pilot study
Tarek Yosef, Wesam Ahmed Ibrahim, Marwa Matboli, Amina Ahmed Swilam, Sarah El-Nakeep
Tarek Yosef, Wesam Ahmed Ibrahim, Sarah El-Nakeep, Gastroenterology and Hepatology Unit, Department of Internal Medicine, Faculty of Medicine, Ain Shams University, Cairo 11591, Egypt
Marwa Matboli, Department of Biochemistry, Faculty of Medicine, Ain Shams University, Cairo 11591, Egypt
Amina Ahmed Swilam, Department of Internal Medicine, Health Affair Directorate, Ministry of Health and Population, Cairo 11591, Egypt
Author contributions: Yosef T supervised the conduction of the study; Ibrahim WA and El-Nakeep S followed the clinical collection of data and the availability of patients; El-Nakeep S and Matboli M formulated the research question and its applicability and wrote the final draft; Matboli M conducted the laboratory analysis; Swilam AA collected the data from the patients; all authors revised and accepted the final submitted manuscript.
Institutional review board statement: The Internal Medicine Department, Faculty of Medicine, Ain Shams University, approved this study’s protocol in 2016 for ethics of conducting the study and in accordance with the ethical standards of the Declaration of Helsinki. Informed consent was obtained from each participant. Both the patients and controls were randomly selected.
Informed consent statement: Informed consent was obtained from each participant. Both the patients and controls were randomly selected.
Conflict-of-interest statement: All Authors declare that they have no conflict of interest.
Data sharing statement: Data sharing of the original excel sheets and other datasets could be obtained upon request and approval of all authors.
STROBE statement: The authors have read the STROBE Statement—checklist of items, and the manuscript was prepared and revised according to the STROBE Statement—checklist of items.
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: Sarah El-Nakeep, MD, Associate Professor, Gastroenterology and Hepatology Unit, Department of Internal Medicine, Faculty of Medicine, Ain Shams University, Ramsees Street, Cairo 11591, Egypt. sarahnakeep@gmail.com
Received: May 4, 2021
Peer-review started: May 4, 2021
First decision: June 16, 2021
Revised: June 21, 2021
Accepted: October 27, 2021
Article in press: October 27, 2021
Published online: December 27, 2021

Stem cell autophagy disruption is responsible for the development of hepatocellular carcinoma (HCC). Many non-coding RNAs are linked to the activation and inhibition of certain genes. The SQSTM1 gene controls stem cell autophagy as shown in previous studies. The upregulation of SQSTM1 is associated with the inhibition of autophagy in cancerous stem cells in patients with HCC.


To determine whether serum microRNA, hsa-miR-519d, is linked to SQSTM1 gene and whether they could be used as diagnostic biomarkers for early-stage HCC.


In silico analysis was performed to determine the most correlated genes of autophagy with microRNAs. SQSTM1 and hsa-miR-519d were validated through this pilot clinical study. This study included 50 Egyptian participants, who were classified into three subgroups: Group 1 included 34 patients with early-stage HCC, Group 2 included 11 patients with chronic liver disease, and Group 3 (control) included 5 healthy subjects. All patients were subjected to full laboratory investigations, including viral markers and alpha-fetoprotein (AFP), abdominal ultrasound, and clinical assessment with the Child–Pugh score calculation. Besides, the patients with HCC underwent triphasic computed tomography with contrast to diagnose and determine the tumor site, size, and number. Quantitative real-time polymerase chain reaction was used to assess hsa-miR-519d-3p and SQSTM1 in the serum of all the study participants.


Hsa-miR-519d-3p was significantly upregulated in patients with HCC compared with those with chronic liver disease and healthy subjects with an area under the curve (AUC) of 0.939, with cutoff value 8.34, sensitivity of 91.2%, and specificity of 81.8%. SQSTM1 was upregulated with an AUC of 0.995, with cutoff value 7.89, sensitivity of 97.1%, and specificity of 100%. AFP significantly increased in patients with HCC with an AUC of 0.794, with cutoff value 7.30 ng/mL, sensitivity of 76.5%, and specificity of 72.7%.


This study is the first to show a direct relation between SQSTM1 and hsa-miR-519d-3p; they are both upregulated in HCC. Thus, they could be used as surrogate diagnostic markers for stem cell autophagy disturbance in early-stage HCC.

Keywords: Autophagy, Hepatocellular carcinoma, miRNA, miR-519d, Stem cell, SQSTM1

Core Tip: Hepatocellular carcinoma (HCC) is the most common primary liver cancer. HCC is associated with poor prognosis due to difficult discovery at an early stage. The molecular pathophysiology behind HCC is not yet fully understood. Autophagy is one of the important affected pathways in HCC pathogenesis. In this study we used in silico analysis to determine a new molecular pathway and find the underlying background controlling genetic and epigenetic pathways. We found that autophagy-controlling gene SQSTM1 is related to hsa-miR-519d-3p. Also, we found that their use as early detecting biomarkers for HCC diagnosis are more efficient than the currently used alpha-fetoprotein.