Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

mRNA Transcriptomic Profiling of Human Hepatocellular Carcinoma Cells HepG2 Treated with Catharanthus roseus-Silver Nanoparticles

Version 1 : Received: 23 September 2021 / Approved: 24 September 2021 / Online: 24 September 2021 (12:44:00 CEST)

How to cite: Azhar, N.A.; Abu Bakar, S.A.; Citartan, M.; Ahmad, N.H. mRNA Transcriptomic Profiling of Human Hepatocellular Carcinoma Cells HepG2 Treated with Catharanthus roseus-Silver Nanoparticles. Preprints 2021, 2021090431 (doi: 10.20944/preprints202109.0431.v1). Azhar, N.A.; Abu Bakar, S.A.; Citartan, M.; Ahmad, N.H. mRNA Transcriptomic Profiling of Human Hepatocellular Carcinoma Cells HepG2 Treated with Catharanthus roseus-Silver Nanoparticles. Preprints 2021, 2021090431 (doi: 10.20944/preprints202109.0431.v1).

Abstract

Background: The demand in the development of cancer nanomedicine has increased due to various limitations in conventional cancer therapy. This study assessed the mRNA transcriptomic profiling of human HepG2 cells exposed to C. roseus-AgNPs. Methods: The proliferative activity of hepatocellular carcinoma (HepG2) and normal human liver (THLE3) cells treated with C. roseus‑AgNPs 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, DEG analysis, GO analysis, and pathway analysis. Results: The mean IC50 values of C. roseus‑AgNPs on HepG2 was 4.38±1.59 µg/mL while on THLE3 cells was 800±1.55 µg/mL. Transciptomic profiling revealed an alteration of 296 genes. C. roseus‑AgNPs induced the expression of stress-associated genes such as MT, HSP and HMOX-1. Cellular signaling pathways were potentially activated through MAPK, TNF and TGF pathways that responsible for apoptosis and cell cycle arrest. The alteration of ARF6, EHD2, FGFR3, RhoA, EEA1, VPS28, VPS25, TSG101 indicated the uptake of C. roseus-AgNPs via both clathrin-dependent and clathrin-independent endocytosis. Conclusions: This study provides the new insights on gene expression study of biosynthesized AgNPs on cancer cells. The cytotoxicity effect is mediated by the aberrant gene alteration, and more interestingly the unique selective antiproliferative properties indicates the C. roseus‑AgNPs as an ideal anticancer candidate.

Keywords

C. roseus; HepG2; silver nanoparticles; AgNPs; mRNA; transcriptomic; gene expression; oxidative stress; apoptosis; cell cycle.

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