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©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
Antihepatoma peptide, scolopentide, derived from the centipede scolopendra subspinipes mutilans
Yu-Xing Hu, Zhuo Liu, Zhen Zhang, Zhe Deng, Zhen Huang, Ting Feng, Qing-Hong Zhou, Si Mei, Chun Yi, Qing Zhou, Pu-Hua Zeng, Gang Pei, Sha Tian, Xue-Fei Tian
Yu-Xing Hu, Zhen Zhang, Zhe Deng, Zhen Huang, Ting Feng, Sha Tian, Xue-Fei Tian, College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Yu-Xing Hu, Xue-Fei Tian, Hunan Key Laboratory of Translational Research in Formulas and Zheng of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Zhuo Liu, Zhen Zhang, Department of Scientific Research, Hunan Academy of Traditional Chinese Medicine Affiliated Hospital, Changsha 410208, Hunan Province, China
Zhe Deng, Hunan Province University Key Laboratory of Oncology of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Qing-Hong Zhou, Department of Pediatric, Shenzhen Hospital of Beijing University of Chinese Medicine, Shenzhen 518000, Guangdong Province, China
Si Mei, Department of Physiology, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Chun Yi, Department of Pathology, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Qing Zhou, Department of Andrology, First Hospital of Hunan University of Chinese Medicine, Changsha 410007, Hunan Province, China
Pu-Hua Zeng, Department of Oncology, Hunan Academy of Traditional Chinese Medicine Affiliated Hospital, Changsha 410208, Hunan Province, China
Gang Pei, College of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Sha Tian, Dr Neher’s Biophysics Laboratory for Innovative Drug Discovery, Macau University of Science and Technology, Macau 999078, China
Author contributions: Tian XF and Tian S contributed equally to this work, and they are co-corresponding authors; Tian XF, Tian S, Pei G, and Hu YX designed the study; Hu YX, Liu Z, Deng Z, Mei S, Yi C, Huang Z, and Feng T performed experiments; Pei G helped perform the high performance liquid chromatography and mass spectrum; Zhang Z performed data analyses; Zhou QH processed figures and tables; Hu YX wrote the manuscript; Tian XF, Zhou Q, and Zeng PH reviewed and edited the manuscript; all authors reviewed and approved the final version of the article.
Supported by the National Natural Science Foundation of China, No. U20A20408 and No. 82074450; Natural Science Foundation of Hunan Province, No. 2020JJ4066 and No. 2021JJ40405; Key scientific research project of Hunan Education Department, No. 21A0243; and Key project of academician workstation guidance project, No. 21YSZQ007.
Institutional animal care and use committee statement: All animal experiments were conducted according to a protocol approved by the Ethics Review Committee of Experimental Animal Welfare at the Hunan University of Chinese Medicine (Approval No. LL2021040705).
Conflict-of-interest statement: The authors declare that they have no competing interests.
Data sharing statement: Dataset available from the corresponding author at 003640@hnucm.edu.cn.
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:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Xue-Fei Tian, PhD, Professor, College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, No. 300 Xueshi Road, Changsha 410208, Hunan Province, China. 003640@hnucm.edu.cn
Received: December 18, 2022
Peer-review started: December 18, 2022
First decision: January 22, 2023
Revised: February 2, 2023
Accepted: March 15, 2023
Article in press: March 15, 2023
Published online: March 28, 2023
Processing time: 100 Days and 8.8 Hours
ARTICLE HIGHLIGHTS
Research background
Centipedes have been used to treat tumors for hundreds of years in China, while current studies rarely focus on hepatoma. The molecular identities of antihepatoma bioactive components in centipedes have not yet been extensively investigated. It is a challenge to isolate and characterize the effective components of centipedes due to limited peptide purification technologies for animal-derived medicines.
Research motivation
The antihepatoma components in centipedes remain unclear. We investigated the centipede components with the strongest antihepatoma activity to develop candidates for antihepatoma drugs.
Research objectives
To purify, characterize, and synthesize the bioactive components with the strongest antihepatoma activity from centipedes and determine the antihepatoma mechanism. To provide a reference for the extraction, purification and characterization of effective components for animal-derived medicines.
Research methods
An antihepatoma peptide (scolopentide) was isolated and identified from the centipede scolopendra subspinipes mutilans using a combination of enzymatic hydrolysis, a Sephadex G-25 column, and two steps of high-performance liquid chromatography. Additionally, the CCK8 assay was used to select the extracted fraction with the strongest antihepatoma activity. The molecular weight of the extracted scolopentide was characterized by quadrupole time-of-flight mass spectrometry, and the sequence was matched by using the Mascot search engine. Scolopentide was then synthesized using solid-phase peptide synthesis methods. The antihepatoma effects of extracted and synthetic scolopentide were confirmed in vitro and in vivo. Mechanistically, flow cytometry and Hoechst staining were conducted to confirm the occurrence of apoptosis. Molecular docking and CCK8 assays were performed to determine the relationship between scolopentide and the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) pathway. Reactive oxygen species assessment, quantitative real-time polymerase chain reaction and Western blot were used to further verify the hypothesis that scolopentide can stimulate the TRAIL pathway to induce apoptosis.
Research results
A small molecule polypeptide with the strongest antihepatoma activity was derived from Scolopendra subspinipes mutilans L. Koch. The molecular weight was 1018.997 Da, and the peptide sequence was RAQNHYCK. Both extracted and synthesized scolopentide had antihepatoma activity in a concentration-dependent manner. Mechanistically, scolopentide activated death receptor 4 (DR4) and DR5 and induced apoptosis in liver cancer cells by promoting the expression of Fas-associated death domain protein (FADD), caspase-8 and caspase-3 through a mitochondria-independent pathway.
Research conclusions
Scolopentide, an antihepatoma peptide, was isolated and identified from centipedes, which activated DR4 and DR5 and induced apoptosis through a mitochondria-independent pathway.
Research perspectives
Scolopentide is considered to be a promising drug candidate for cancer treatment, especially treatment of liver cancer. Ways in which to modify the spatial architecture of synthetic scolopentide, fully activate the TRAIL pathway and improve its antihepatoma activity will be key areas for future research.
