Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Oct 28, 2023; 29(40): 5543-5556
Published online Oct 28, 2023. doi: 10.3748/wjg.v29.i40.5543
Roles of phosphatidylinositol-3-kinases signaling pathway in inflammation-related cancer: Impact of rs10889677 variant and buparlisib in colitis-associated cancer
Nurul Nadirah Razali, Raja Affendi Raja Ali, Khairul Najmi Muhammad Nawawi, Azyani Yahaya, Norshafila Diana Mohd Rathi, Norfilza Mohd Mokhtar
Nurul Nadirah Razali, Norshafila Diana Mohd Rathi, Norfilza Mohd Mokhtar, Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
Raja Affendi Raja Ali, School of Medical and Life Sciences, Sunway University, Sunway City 47500, Malaysia
Raja Affendi Raja Ali, Khairul Najmi Muhammad Nawawi, Norfilza Mohd Mokhtar, GUT Research Group, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
Raja Affendi Raja Ali, Khairul Najmi Muhammad Nawawi, Gastroenterology Unit, Department of Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
Azyani Yahaya, Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
Author contributions: Mokhtar NM and Raja Ali RA contributed to the conceptualization; Mokhtar NM, Raja Ali RA, Muhammad Nawawi KN, and Razali NN contributed to the methodology and participant recruitments; Razali NN, and Mohd Rathi ND contributed to the animal handling; Razali NN, Yahaya A, and Mokhtar NM performed the validation; Razali NN, and Mokhtar NM contributed to the data analysis; Razali NN wrote the original draft preparation; Mokhtar NM, Raja Ali RA, and Muhammad Nawawi KN wrote the review and editing; Mokhtar NM, Raja Ali RA, and Muhammad Nawawi KN contributed to the supervision; All authors have read and agreed to the published version of the manuscript.
Supported by The Fundamental Research Grant Scheme, Ministry of Higher Education, Malaysia, No. FRGS/1/2018/SKK06/UKM/02/4.
Institutional review board statement: The human study has been approved under the Universiti Kebangsaan Malaysia Research Ethics Committee with a reference number: UKM/PPI/111/8/JEP-2019-572.
Institutional animal care and use committee statement: The animal study has been approved under the Animal Ethics Committee of Universiti Kebangsaan Malaysia with a reference number: PPUKM/2019/NORFILZA/25-SEPT./1035-SEPT.-2019-DEC.-2021 has approved the animal handling protocol.
Informed consent statement: Written informed consent was obtained from all the patients.
Conflict-of-interest statement: There is no conflict of interest.
Data sharing statement: Our raw data will be shared upon official request.
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: Norfilza Mohd Mokhtar, PhD, Professor, Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras 56000, Kuala Lumpur, Malaysia. norfilza@ppukm.ukm.edu.my
Received: July 20, 2023
Peer-review started: July 20, 2023
First decision: August 25, 2023
Revised: September 5, 2023
Accepted: October 11, 2023
Article in press: October 11, 2023
Published online: October 28, 2023
Abstract
BACKGROUND

Phosphatidylinositol-3-kinases (PI3K) is a well-known route in inflammation-related cancer. Recent discovery on PI3K-related genes revealed a potential variant that links ulcerative colitis (UC) and colorectal cancer (CRC) with colitis-associated cancer (CAC). PI3K/AKT pathway has been recommended as a potential additional therapeutic option for CRC due to its substantial role in modifying cellular processes. Buparlisib is a pan-class I PI3K inhibitor previously shown to reduce tumor growth.

AIM

To investigate the regulation of rs10889677 and the role of buparlisib in the PI3K signaling pathway in CAC pathogenesis.

METHODS

Genomic DNA from 32 colonic samples, including CAC (n = 7), UC (n = 10) and CRC (n = 15), was sequenced for the rs10889677 mutation. The mutant and wildtype fragments were amplified and cloned in the pmirGLO vector. The luciferase activity of cloned vectors was assessed after transfection into the HT29 cell line. CAC mice were induced by a mixture of a single azoxymethane injection and three cycles of dextran sulphate sodium, then buparlisib was administered after 14 d. The excised colon was subjected to immunohistochemistry for Ki67 and Cleaved-caspase-3 markers and quantitative real-time polymerase chain reaction analysis for Pdk1 and Sgk2.

RESULTS

Luciferase activity decreased by 2.07-fold in the rs10889677 mutant, confirming the hypothesis that the variant disrupted miRNA binding sites, which led to an increase in IL23R expression and the activation of the PI3K signaling pathway. Furthermore, CAC-induced mice had a significantly higher disease activity index (P < 0.05). Buparlisib treatment significantly decreased mean weight loss in CAC-induced mice (P < 0.05), reduced the percentage of proliferating cells by 5%, and increased the number of apoptotic cells. The treatment also caused a downward trend of Pdk1 expression and significantly decreased Sgk2 expression.

CONCLUSION

Our findings suggested that the rs10889677 variant as a critical initiator of the PI3K signaling pathway, and buparlisib had the ability to prevent PI3K-non-AKT activation in the pathophysiology of CAC.

Keywords: Colitis-associated cancer, Colorectal cancer, Phosphatidylinositol 3-kinase, Animal model, Luciferases, Renilla, Phosphatidylinositol 3-kinase inhibitor

Core Tip: The role of phosphatidylinositol-3-kinases (PI3K) in promoting cancer progression has been widely acknowledged due to its crucial involvement in regulating the survival, differentiation, and proliferation of cancer cells. Here, we investigate the role of PI3K signaling in colitis-associated cancer (CAC) pathogenesis by studying the regulation of potential variant in PI3K-related gene in the colorectal cancer cell line and the utilization of PI3K inhibitor, buparlisib, in the CAC-induced mice model. We suggested that rs10889677 variant plays a crucial role in initiating the PI3K signaling pathway, and buparlisib has the capability to inhibit PI3K-non-AKT activation in the pathophysiology of CAC.