Graphene Oxide Nanofilm and Chicken Embryo Extract Decrease the Invasiveness of HepG2 Liver Cancer Cells

Abstract

Background: The extracellular matrix (ECM) is a mosaic of various structural and functional proteins that cooperate with the cell, regulate adhesion, and consequently manage its further fate. Liver destruction is accompanied by a disruption of the physicochemical properties of the ECM which deregulates the cell-ECM interaction and can lead to uncontrolled proliferation and neoplastic transformation of cells. Therefore, it can be assumed that ECM modification and restoration of its characteristics for healthy tissue may counteract uncontrolled cell proliferation. The purpose of the presented research model was to optimise the physical characteristics of ECM by introducing a graphene oxide plane/nanofilm (nfGO) and enriching the cell environment with potentially missing proteins by adding a functional protein cocktail (chicken embryo liver extract) and determine the impact of these factors on cell-ECM cooperation and its consequences on adhesion, proliferation, and cell phase, which are factors of the invasiveness of cancer cells.Results: Experiments were performed with non-cancer HS-5 cells and liver cancer cells HepG2 and C3A. The cells were divided into four groups; (1) control, (2) cultured on GO nanofilm, (3) cultured with the addition of chicken embryo liver extract (CELE) to the medium and (4) cultured on the GO nanofilm with the addition of CELE. CELE contained 1735 proteins; the top 57 of these proteins have been presented. The use of GO nanofilm as well as CELE and nfGO + CELE reduced the proliferation of HepG2 cancer cells to the greatest extent; this is in contrast to non-cancer cells and also to C3A cancer cells. Furthermore, the combined use of the CELE protein cocktail and GO substrate effectively resulted in a decrease in the population of HepG2 cells in the G0/G1 phase and an increase of the population in G2/M. Molecular analysis of HepG2 cancer cells also showed an increase in the expression of genes responsible for adhesion such as fak (focal adhesion kinase), e-cadherin, and n-cadherin and a decrease in β-catenin, which is considered a proto-oncogen. Conclusions: Studies have shown that both the GO surface structure on which the cells are grown as well as the presence of a multi-component natural cocktail of regulatory proteins, can modify the expression of integrins, increase adhesion and, as a consequence, proliferation and the cell cycle - entering the resting phase. For the first time, it has been documented that hepatic cancer cells of the HepG2 line under the influence of stimuli derived from mimic ECM (graphene oxide) in interaction with a unique protein complex derived from chicken liver embryo decreased of the invasiveness of cancer cells.

Design Strategies of Tumor‐Targeted Delivery Systems Based on 2D Nanomaterials

“…Sosnowska et al added a functional protein cocktail, chicken embryo liver extract protein cocktail, and GOx substrate to a GO plane to increase the expression of adhesion-related genes and decrease the expression of the protooncogene β-catenin to decrease the invasiveness of HepG2 liver cancer cells. [156] On the one hand, the simulated ECM skeleton prepared using the BP photothermal effect showed good photothermal ablation of tumors. On the other hand, released bioactive substances promoted fibroblast proliferation, promoted the expression of genes related to angiogenesis and ECM components of skin tissue regeneration, and accelerated wound healing (Figure 10e).…”