Anticancer Molecular Mechanism of Protocatechuic Acid Loaded on Folate Coated Functionalized Graphene Oxide Nanocomposite Delivery System in Human Hepatocellular Carcinoma

Abstract

Liver cancer is listed as the fifth-ranked cancer, responsible for 9.1% of all cancer deaths globally due to its assertive nature and poor survival rate. To overcome this obstacle, efforts have been made to ensure effective cancer therapy via nanotechnology utilization. Recent studies have shown that functionalized graphene oxide (GO)-loaded protocatechuic acid has shown some anticancer activities in both passive and active targeting. The nanocomposites’ physicochemical characterizations were conducted. A lactate dehydrogenase experiment was conducted to estimate the severity of cell damage. Subsequently, a clonogenic assay was carried out to examine the colony-forming ability during long-term exposure of the nanocomposites. The Annexin V/ propidium iodide analysis showed that nanocomposites induced late apoptosis in HepG2 cells. Following the intervention of nanocomposites, cell cycle arrest was ascertained at G2/M phase. There was depolarization of mitochondrial membrane potential and an upregulation of reactive oxygen species when HepG2 cells were induced by nanocomposites. Finally, the proteomic profiling array and quantitative reverse transcription polymerase chain reaction revealed the expression of pro-apoptotic and anti-apoptotic proteins induced by graphene oxide conjugated PEG loaded with protocatechuic acid drug folic acid coated nanocomposite (GOP–PCA–FA) in HepG2 cells. In conclusion, GOP–PCA–FA nanocomposites treated HepG2 cells exhibited significant anticancer activities with less toxicity compared to pristine protocatechuic acid and GOP–PCA nanocomposites, due to the utilization of a folic acid-targeting nanodrug delivery system.

Graphene Oxide Loaded with Protocatechuic Acid and Chlorogenic Acid Dual Drug Nanodelivery System for Human Hepatocellular Carcinoma Therapeutic Application

“…The nanoformulation of dual drug-loaded graphene oxide nanocomposites were successfully formulated in this study. Graphene oxide is a hydrophilic nanocarrier which possess characteristics such as a large surface area, biocompatibility, bioavailability, endocytosis ability, mechanical properties and low cytotoxicity [ 23 ]. The attachment of the functional groups on the basal and edge of the GO makes it a suitable nanocarrier for multiple drug attachment and modifications for the nanodrug delivery system.…”

“…Figure 3 d represents the particle size distribution of GOP-PCA/CA whereas Figure 3 e represents the particle size distribution of GOP-PCA/CA-FA nanocomposite. Overall, the synthesized nanocomposites show a spherical shape, narrow size distribution and a smooth surface [ 23 , 33 ].…”

“…In vitro release studies of PCA and CA from GOP-PCA/CA and GOP-PCA/CA-FA nanocomposites were conducted in a human body replicated phosphate buffer saline (PBS) solution of pH 4.8 (intercellular lysosomal pH) and pH 7.4 (human blood pH) at 37 °C [ 23 ]. The absorption for PCA and CA were taken at 256 nm and 330 nm, respectively, as these are their individual lambda maxes [ 8 ].…”

“…The activated carboxylic acid group in GO was catalyzed using 400 mg of N-(3-Dimethylaminopropyl-N’-ethylcarbodiimide hydrochloride (EDC) and 240 mg of N-hydroxysulfosuccinimide (NHS) and stirred for 24 h. This was followed by 1.5 g of PEG4000, which was added to the above suspension and constantly stirred overnight. Finally, the suspension was washed with deionized water and centrifuged to obtain GO conjugated PEG (GOP) [ 23 ].…”

“…Whereas, overexpression of receptor on the cancer cell membrane assists in ligand-based active targeting [ 22 ]. The multidrug delivery of passive and active targeting therapy has shown enhanced anticancer effects in the clinical application [ 23 ]. In this study, we have described the formation of a graphene oxide nanocomposite by the encapsulation of two drugs via nanoabsorption, incorporated with passive and active targeting for a nanodrug delivery system.…”