Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 28, 2022; 28(32): 4620-4634
Published online Aug 28, 2022. doi: 10.3748/wjg.v28.i32.4620
Anti-tumour activity and toxicological studies of combination treatment of Orthosiphon stamineus and gemcitabine on pancreatic xenograft model
Ashwaq Hamid Salem Yehya, Ayappa V Subramaniam, Muhammad Asif, Gurjeet Kaur, Amin M S Abdul Majid, Chern Ein Oon
Ashwaq Hamid Salem Yehya, Ayappa V Subramaniam, Gurjeet Kaur, Chern Ein Oon, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang 11800, Malaysia
Ashwaq Hamid Salem Yehya, Cancer Research, Eman Biodiscoveries, Kedah 08000, Malaysia
Muhammad Asif, Department of Pharmacology, Faculty of Pharmacy, The Islamia University of Bahawalpur, Punjab 63100, Pakistan
Amin M S Abdul Majid, Department of Cancer Biology and Therapeutics, The John Curtin School of Medical Research, Australia 2601, Australia
Author contributions: Yehya AHS and Subramaniam AV carried out the experiments; Yehya AHS drafted the manuscript; Yehya AHS, Asif M, and Kaur G analysed the data; Abdul Majid AMS conceived and planned the experiments; Oon CE designed the experiments, derived the model, and supervised the project; All authors have read and approved the final manuscript.
Supported by the NKEA Research Grant Scheme (NRGS) by the Ministry of Agriculture Malaysia, No. 304/CIPPM/650736/k123.
Institutional animal care and use committee statement: The animal study was approved and conducted under strict guidelines according to USM Animal Ethics Committee (Reference #: USM/Animal Ethics Approval/2016/(97) (746).
Conflict-of-interest statement: Amin Malik Shah Abdul Majid has a commercial interest in C5EOSEW5050ESA O.s extract of NuvastaticTM. All the other authors declare no financial and non-financial competing interests.
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: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Chern Ein Oon, PhD, Associate Professor, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Chancellory II, Penang 11800, Malaysia. chern.oon@usm.my
Received: January 10, 2022
Peer-review started: January 10, 2022
First decision: April 16, 2022
Revised: April 26, 2022
Accepted: June 26, 2022
Article in press: June 26, 2022
Published online: August 28, 2022
Processing time: 227 Days and 17.9 Hours
Abstract
BACKGROUND

Pancreatic cancer is the most aggressive cancer type. Gemcitabine is the first line chemo-drug used for pancreatic cancer but exerts a broad spectrum of organ toxicities and adverse effects in patients.

AIM

To evaluate the anti-tumour activity and toxicological effects of Orthosiphon stamineus extract formulation (ID: C5EOSEW5050ESA trademarked as Nuva-staticTM), and gemcitabine combination on pancreatic xenograft model.

METHODS

Mice were randomly divided into six groups of 6 mice each (n = 6) and given different treatments for 28 d. The study design consisted of a 2 x 3 factorial treatment structure, with gemcitabine (yes/no) by oral (at 1200 and 400 mg/kg per day). Human pancreatic cancer cells were injected subcutaneously into the flanks of athymic nude mice. C5EOSEW5050ESA (200 or 400 mg/kg per day) was administered orally, while gemcitabine (10 mg/kg per 3 d) was given intraperitoneally either alone or in combination treatment. Histopathological analyses of vital organs, tumour tissues, and incidence of lethality were analysed. Analyses of tumour necrosis and proliferation were determined by haematoxylin-eosin staining and immunohistochemistry for Ki-67, respectively.

RESULTS

No signs of toxicity or damage to vital organs were observed in all treatment groups compared to the untreated group. C5EOSEW5050ESA at 200 mg/kg and gemcitabine combination had no additive antitumor effects compared to a single treatment. Remarkably, a comparably greater response in a reduction in tumour growth, Ki-67 protein expression, and necrosis was demonstrated by 400 mg/kg of C5EOSEW5050ESA and gemcitabine combination than that of the individual agents.

CONCLUSION

These results highlighted the synergistic activity of C5EOSEW5050ESA with gemcitabine to reduce pancreatic tumour growth in mice compared to a single treatment. Thus, this study provides valuable insights into using C5EOSEW5050ESA as a complementary treatment with gemcitabine for pancreatic cancer.

Keywords: Pancreatic cancer; Orthosiphon stamineus; C5EOSEW5050ESA; Gemcitabine; Complementary medicine

Core Tip: The combination of C5EOSEW5050ESA at 400 mg/kg and gemcitabine synergistically reduced tumour growth compared to either drug alone by reducing Ki-67 cell proliferation marker and tumour necrosis. In addition, no signs of toxicity or damage to vital organs in single C5EOSEW5050ESA or gemcitabine and chemo-herbal combinations treated animals compared to the untreated group, indicating the safety and efficacy of the combination treatment in a short term study. Findings from this study may provide the basis for product formulation and manufacturing of botanical drugs to be used as complementary medicine for the treatment of cancer.