Observational Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Oncol. Aug 15, 2022; 14(8): 1574-1584
Published online Aug 15, 2022. doi: 10.4251/wjgo.v14.i8.1574
RASSF1A methylation as a biomarker for detection of colorectal cancer and hepatocellular carcinoma
Jian Li, Huan Li, Zeng-Ci Run, Zhen-Lei Wang, Tao Jiang, Yang An, Zhi Li
Jian Li, Zeng-Ci Run, Zhen-Lei Wang, Zhi Li, Department of General Surgery, Affiliated Tumor Hospital of Zhengzhou University, Henan Tumor Hospital, Zhengzhou 450000, Henan Province, China
Huan Li, Department of Gastroenterology, Sixth Medical Center of Chinese PLA General Hospital, Beijing 100048, China
Tao Jiang, Medicine Innovation Research Division of Chinese PLA General Hospital, Beijing 100853, China
Yang An, Faculty of Hepato-Pancreato-Biliary Surgery, Sixth Medical Center of Chinese PLA General Hospital, Beijing 100048, China
Author contributions: Li J and Li Z designed the study; Li J and Li H contributed equally to this study; An Y and Li Z are the co-corresponding authors; Run ZC, Wang ZL, Jiang T, and An Y collected the samples; Li J, Li H, An Y performed the research; Li J, Li H and Li Z analyzed the date; Li J wrote the paper; Li J and Li Z revised the manuscript for final submission.
Supported by National Natural Science Foundation of China, No. 81972010; Science Developing Funds of Navy General Hospital, No. CXPY201810.
Institutional review board statement: The study was reviewed and approved by the Chinese PLA General Hospital Review Board.
Informed consent statement: All study participants or their legal guardian provided written informed consent prior to study enrollment.
Conflict-of-interest statement: We declare that we have no financial or personal relationships with other individuals or organizations that can inappropriately influence our work and that there is no professional or other personal interest of any nature in any product, service and/or company that could be construed as influencing the position presented in or the review of the manuscript.
Data sharing statement: No additional data are available.
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: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Zhi Li, DA, Chief Doctor, Department of General Surgery, Affiliated Tumor Hospital of Zhengzhou University, Henan Tumor Hospital, No. 127 Dong Ming Road, Jin Shui District, Zhengzhou 450000, Henan Province, China. lizhihn2009@163.com
Received: May 26, 2022
Peer-review started: May 26, 2022
First decision: June 21, 2022
Revised: July 2, 2022
Accepted: July 22, 2022
Article in press: July 22, 2022
Published online: August 15, 2022
Processing time: 76 Days and 3.7 Hours
Abstract
BACKGROUND

Studies have validated the potential of methylated cell-free DNA as a biomarker in various tumors, and methylated DNA in plasma may be a potential biomarker for cancer.

AIM

To evaluate the diagnostic value of RASSF1A methylation in plasma for colorectal cancer (CRC) and hepatocellular carcinoma (HCC).

METHODS

A total of 92 CRC patients, 67 colorectal polyp (CRP) patients, 63 HCC patients, and 66 liver cirrhosis (LC) patients were enrolled. The plasma DNA was subjected to DNA extraction, double-strand DNA concentration determination, bisulfite conversion, purification, single-strand DNA concentration determination, and digital polymerase chain reaction (PCR) detection. The methylation rate was calculated. The diagnostic value was evaluated by the area under the curve (AUC).

RESULTS

The age and sex in the CRC and CRP groups and the HCC and LC groups were also matched. The DNA methylation rate of RASSF1A in plasma in the CRC group was 2.87 ± 1.80, and that in the CRP group was 1.50 ± 0.64. DNA methylation of RASSF1A in plasma showed a significant difference between the CRC and CRP groups. The AUC of RASSF1A methylation for discriminating the CRC and CRP groups was 0.82 (0.76-0.88). The AUCs of T1, T2, T3 and T4 CRC and CRP were 0.83 (0.72-0.95), 0.87 (0.78-0.95), 0.86 (0.77-0.95), and 0.75 (0.64-0.85), respectively. The DNA methylation rate of RASSF1A in plasma in the HCC group was 4.45 ± 2.93, and that in the LC group was 2.46 ± 2.07. DNA methylation of RASSF1A in plasma for the HCC and LC groups showed a significant difference. The AUC of RASSF1A methylation for discriminating the HCC and LC groups was 0.70 (0.60-0.79). The AUCs of T1, T2, T3 and T4 HCC and LC were 0.80 (0.61, 1.00), 0.74 (0.59-0.88), 0.60 (0.42-0.79), and 0.68 (0.53-0.82), respectively.

CONCLUSION

RASSF1A methylation in plasma detected by digital PCR may be a potential biomarker for CRC and HCC.

Keywords: RASSF1A; Methylation; Digital polymerase chain reaction; Colorectal cancer; Hepatocellular carcinoma

Core Tip: Accurate quantitative polymerase chain reaction (qPCR) quantification relies on a standard curve and good amplification efficiency and is sensitive to factors affecting amplification efficiency. Digital PCR technology is an emerging PCR technology. In this study, we evaluated the diagnostic value of RASSF1A methylation in plasma by digital polymerase chain reaction.