Case Control Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Nov 14, 2019; 25(42): 6322-6341
Published online Nov 14, 2019. doi: 10.3748/wjg.v25.i42.6322
MicroRNA signature in patients with hepatocellular carcinoma associated with type 2 diabetes
Moustafa Nouh Elemeery, Marwa Anwar Mohamed, Marwa Ahmed Madkour, Mohammed Mohammed Shamseya, Noha Mahmoud Issa, Ahmed Noah Badr, Doaa Ahmed Ghareeb, Cheol-Ho Pan
Moustafa Nouh Elemeery, Département de Neurosciences, CRCHUM, Université de Montréal, Montréal, Quebec H2X 3E4, Canada
Moustafa Nouh Elemeery, Medical Biotechnology Laboratory, Genetic Engineering and Biotechnology Research Division, National Research Centre, Cairo 12622, Egypt
Moustafa Nouh Elemeery, Cheol-Ho Pan, Natural Product Informatics Research Center, KIST Gangneung Institute of Natural Products, Gangneung 25451, South Korea
Moustafa Nouh Elemeery, Cheol-Ho Pan, Division of Bio-Medical Science and Technology, KIST School, Korea University of Science and Technology, Seoul 02792, South Korea
Marwa Anwar Mohamed, Department of Chemical Pathology, Medical Research Institute, Alexandria University, Alexandria 21511, Egypt
Marwa Ahmed Madkour, Mohammed Mohammed Shamseya, Experimental and Clinical Internal Medicine Department, Medical Research Institute, Alexandria University, Alexandria 21511, Egypt
Noha Mahmoud Issa, Human Genetics Department, Medical Research Institute, Alexandria University, Alexandria 21511, Egypt
Ahmed Noah Badr, Food Toxicology and Contaminates Department, National Research Centre, Dokki, Cairo 12622, Egypt
Doaa Ahmed Ghareeb, Bioscreening and preclinical trial lab, Biochemistry Department, Faculty of Science, Alexandria University, Alexandria 12522, Egypt
Doaa Ahmed Ghareeb, Pharmaceutical and fermentation industries development center, the city of scientific research and technological applications, Alexandria 26411, Egypt
Author contributions: Elemeery MN designed the research, performed the laboratory work, and drafted the manuscript; Mohamed MA and Issa NM supplied reagents and performed the laboratory work; Madkour MA performed the case selection and the clinical and radiological evaluations, and drafted the manuscript; Shamseya MM performed the case selection and the clinical and radiological evaluations; Badr AN performed the data analysis and biological parameter assessment; Ghareeb DA performed the RT-qPCR work; Pan CH supplied reagents and designed the primers; All authors contributed equally to the manuscript revision.
Institutional review board statement: The study was approved by the ethical committees of the Medical Research Institute of Alexandria University; the National Hepatology and Tropical Medicine Research Institute (NHTMRI, Giza, Egypt); and the National Research Centre (NRC, Cairo, Egypt).
Informed consent statement: All patients provided informed consent.
Conflict-of-interest statement: No benefits in any form have been received or will be received from a commercial party related directly or indirectly to the subject of this article.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at
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 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:
Corresponding author: Cheol-Ho Pan, PhD, Professor, Natural Product Informatics Research Center, KIST Gangneung Institute of Natural Products, Saimdang-ro, Gangneung 25451, South Korea.
Telephone: +82-10-2207-9702
Received: September 3, 2019
Peer-review started: September 3, 2019
First decision: October 14, 2019
Revised: October 29, 2019
Accepted: November 7, 2019
Article in press: November 7, 2019
Published online: November 14, 2019

Nonalcoholic steatohepatitis-related cirrhosis is one of the liver complications in type 2 diabetes mellitus (T2DM) and reported to be a risk factor for developing hepatocellular carcinoma (HCC). A reliable screening biomarker of liver cirrhosis (LC) and HCC among T2DM patients is important to reduce the morbidity and mortality of this disease. MicroRNA (miRNA) is considered a key player in HCC and T2DM, and it might be a hidden culprit in diabetes-associated HCC, making it a promising reliable prognostic tool.


To investigate the signature of serum miRNAs as early biomarkers for the screening of HCC among diabetic patients.


Expression profiles of miRNAs in serum samples of diabetic LC and diabetic HCC patients were assessed using Illumina sequencing; then, RT-qPCR was used to validate significantly altered miRNAs between the two groups. Candidate miRNAs were tested in serum samples of 200 T2DM patients, 270 LC patients, 200 HCC patients, and 225 healthy control subjects. Additionally, receiver operating characteristic (ROC) analysis, with area under the curve (AUC), was performed to assess the diagnostic performance of the screened miRNAs for discriminating HCC from LC and nonmalignant patients (LC + T2DM).


Expression of the sequenced miRNAs in serum was different in HCC vs LC-positive T2DM patients. Two miRNAs (miR-34a, miR-221) were significantly up-regulated and five miRNAs (miR-16, miR-23-3p, miR-122-5p, miR-198, miR-199a-3p) were significantly down-regulated in HCC compared to LC patients. Analysis of ROC curve demonstrated that the combination of these seven miRNAs can be used as a reliable biomarker for detection of HCC in diabetic patients, as it could identify HCC with high diagnostic accuracy in diabetic LC patients (AUC = 0.993) and in diabetic nonmalignant patients (AUC = 0.961).


This study validates a panel of serum miRNAs that can be used as a reliable noninvasive screening biomarker of HCC among T2DM cirrhotic and noncirrhotic patients. The study recommends further research to shed light on a possible role of c-Met in T2DM-associated HCC via the miRNA regulatory pathway.

Keywords: MicroRNA, Hepatocellular carcinoma, Type 2 diabetes, Nonalcoholic fatty liver disease

Core tip: The identification of high-sensitivity biomarkers for detection of he-patocellular carcinoma (HCC) in high-risk individuals is essential. The present study was undertaken to identify and validate serum microRNAs as potential biomarkers for HCC among type 2 diabetes patients.