Basic Study
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World J Hepatol. Mar 27, 2023; 15(3): 393-409
Published online Mar 27, 2023. doi: 10.4254/wjh.v15.i3.393
mRNA transcriptome profiling of human hepatocellular carcinoma cells HepG2 treated with Catharanthus roseus-silver nanoparticles
Nur Asna Azhar, Siti Aishah Abu Bakar, Marimuthu Citartan, Nor Hazwani Ahmad
Nur Asna Azhar, Siti Aishah Abu Bakar, Marimuthu Citartan, Nor Hazwani Ahmad, Department of Biomedical Science, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Kepala Batas 13200, Pulau Pinang, Malaysia
Nur Asna Azhar, Nor Hazwani Ahmad, Liver Malignancies Research Program, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Kepala Batas 13200, Pulau Pinang, Malaysia
Siti Aishah Abu Bakar, Faculty of Bioresources and Food Industry, Universiti Sultan Zainal Abidin, Besut Campus, Besut 22200, Terengganu, Malaysia
Author contributions: Ahmad NH contributed to the methodology; Ahmad NH contributed to the validation; Azhar NA contributed to the conceptualisation; Azhar NA and Abu Bakar SA contributed to the formal analysis; Azhar NA contributed to the investigation; Ahmad NH contributed to the resources; Azhar NA, Abu Bakar SA, Citartan M and Ahmad NH contributed to the data curation; Azhar NA contributed to original draft preparation; Ahmad NH and Citartan M contributed to the review and editing; Ahmad NH and Citartan M contributed to the supervision; Ahmad NH contributed to project administration; Ahmad NH contributed to funding acquisition; all authors have read and agreed to the published version of the manuscript.
Supported by Fundamental Research Grant Scheme from the Malaysian Ministry of Higher Education, No. FRGS/1/2015/SG03/USM/03/1.
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: No additional data are available.
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: Nor Hazwani Ahmad, PhD, Senior Lecturer, Department of Biomedical Science, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Jalan Tun Hamdan Sheikh Tahir, Kepala Batas 13200, Pulau Pinang, Malaysia. norhazwani@usm.my
Received: October 27, 2022
Peer-review started: October 27, 2022
First decision: December 31, 2022
Revised: January 17, 2023
Accepted: March 3, 2023
Article in press: March 3, 2023
Published online: March 27, 2023
Processing time: 146 Days and 5.3 Hours
Abstract
BACKGROUND

The demand for the development of cancer nanomedicine has increased due to its great therapeutic value that can overcome the limitations of conventional cancer therapy. However, the presence of various bioactive compounds in crude plant extracts used for the synthesis of silver nanoparticles (AgNPs) makes its precise mechanisms of action unclear.

AIM

To assessed the mRNA transcriptome profiling of human HepG2 cells exposed to Catharanthus roseus G. Don (C. roseus)-AgNPs.

METHODS

The proliferative activity of hepatocellular carcinoma (HepG2) and normal human liver (THLE3) cells treated with C. roseusAgNPs were measured using MTT assay. The RNA samples were extracted and sequenced using BGIseq500 platform. This is followed by data filtering, mapping, gene expression analysis, differentially expression genes analysis, Gene Ontology analysis, and pathway analysis.

RESULTS

The mean IC50 values of C. roseusAgNPs on HepG2 was 4.38 ± 1.59 μg/mL while on THLE3 cells was 800 ± 1.55 μg/mL. Transcriptome profiling revealed an alteration of 296 genes. C. roseusAgNPs induced the expression of stress-associated genes such as MT, HSP and HMOX-1. Cellular signalling pathways were potentially activated through MAPK, TNF and TGF pathways that are responsible for apoptosis and cell cycle arrest. The alteration of ARF6, EHD2, FGFR3, RhoA, EEA1, VPS28, VPS25, and TSG101 indicated the uptake of C. roseus-AgNPs via both clathrin-dependent and clathrin-independent endocytosis.

CONCLUSION

This study provides new insights into gene expression study of biosynthesised AgNPs on cancer cells. The cytotoxicity effect is mediated by the aberrant gene alteration, and more interestingly the unique selective antiproliferative properties indicate the C. roseusAgNPs as an ideal anticancer candidate.

Keywords: Catharanthus roseus; HepG2; Silver nanoparticles; Transcriptome; oxidative stress; Apoptosis; Cell cycle

Core Tip: Despite the increased attention on cancer nanomedicine which is advantageous to overcome the limitations of conventional cancer treatment, the information on the selectivity and detailed mechanisms at the cellular and molecular level remain unclear. To evaluate its selectivity effects, the proliferative activity of both liver cancer cells HepG2 and normal liver cells THLE-3 in response to Catharanthus roseus-silver nanoparticles (C. roseus-AgNPs) was assessed. To determine the possible signalling pathways induced by the C. roseus-AgNPs, the mRNA transcriptome profiling of hepatocellular carcinoma cell line HepG2 was performed, highlighting the expression of genes associated with oxidative stress, apoptosis, and cell cycle arrest. The elucidation of its selectivity effects and detailed wide genome screening would enlighten the cellular and molecular signalling pathways and provide a strong basis towards the development of C. roseus-AgNPs as an anticancer drug for liver cancer.