Gemcitabine and Antisense-microRNA Co-encapsulated PLGA–PEG Polymer Nanoparticles for Hepatocellular Carcinoma Therapy

Abstract

Hepatocellular carcinoma (HCC) is highly prevalent, and the third most common cause of cancer-associated deaths worldwide. HCC tumors respond poorly to chemotherapeutic anticancer agents due to inherent and acquired drug resistance, and low drug permeability. Targeted drug delivery systems with significant improvement in therapeutic efficiency are needed for successful HCC therapy. Here, we report the results of a technique optimized for the synthesis and formulation of antisense-miRNA-21 and gemcitabine (GEM) co-encapsulated PEGylated-PLGA nanoparticles (NPs) and their in vitro therapeutic efficacy in human HCC (Hep3B and HepG2) cells. Water-in-oil-in-water (w/o/ w) double emulsion method was used to coload antisense-miRNA-21 and GEM in PEGylated-PLGA-NPs. The cellular uptake of NPs displayed time dependent increase of NPs concentration inside the cells. Cell viability analyses in HCC (Hep3B and HepG2) cells treated with antisense-miRNA-21 and GEM co-encapsulated NPs demonstrated a nanoparticle concentration dependent decrease in cell proliferation, and the maximum therapeutic efficiency was attained in cells treated with nanoparticles co-encapsulated with antisense-miRNA-21 and GEM. Flow cytometry analysis showed that control NPs and antisense-miRNA-21-loaded NPs are not cytotoxic to both HCC cell lines, whereas treatment with free GEM and GEM-loaded NPs resulted in ~9% and ~15% apoptosis, respectively. Cell cycle status analysis of both cell lines treated with free GEM or NPs loaded with GEM or antisense-miRNA-21 displayed a significant cell cycle arrest at the S-phase. Cellular pathway analysis indicated that Bcl2 expression was significantly upregulated in GEM treated cells, and as expected, PTEN expression was noticeably upregulated in cells treated with antisense-miRNA-21. In summary, we successfully synthesized PEGylated-PLGA nanoparticles co- encapsulated with antisense-miRNA-21 and GEM. These co-encapsulated nanoparticles revealed increased treatment efficacy in HCC cells, compared to cells treated with either antisense-miRNA-21- or GEM-loaded NPs at equal concentration, indicating that down-regulation of endogenous miRNA-21 function can reduce HCC cell viability and proliferation in response to GEM treatment.

M6A-mediated upregulation of LINC00958 increases lipogenesis and acts as a nanotherapeutic target in hepatocellular carcinoma

“…PEGylated NPs are regarded as "stealth NPs" and characterized by increased circulation time in vivo and tumor uptake. The surface shielding with PEG avoids plasma protein adsorption, protects NPs from the immune recognition, and increases bioavailability [18].…”

“…Poly(ethylene glycol) (PEG) is safe for clinical application and has been used in many Food and Drug Administration-approved medications including intravenous injections [17]. PEGylated PLGA NPs have been acknowledged as one of the best controlled release nanoplatforms for targeted drug delivery [18].…”

Utilização de Nanopartículas no Tratamento do Câncer: Aspectos Gerais, Mecanismos de Ação Antineoplásicos e Aplicabilidades Tumorais

“…PLA e PEG também podem ser usados como carreadores de moléculas, como miRNA, conforme demonstrado no experimento de Devulapally et al 55 . O Quadro 2 mostra o resumo das principais NP abordadas e suas utilizações gerais que encapsularam miRNA supressor de proteína e a droga gentamicina contra células de hepatocarcinoma em NP de PLA.…”

Dual tumor-targeted poly(lactic-<em>co</em>-glycolic acid)&ndash;polyethylene glycol&ndash;folic acid nanoparticles: a novel biodegradable nanocarrier for secure and efficient antitumor drug delivery

“…As the most attractive and efficient polymeric nanocarrier, PLGA nanoparticles are widely used in clinical applications. [28][29][30][31][32] Additionally, polyethylene glycolated (PEGylated) PLGA nanoparticles have been developed to improve bioavailability and antitumor targeting further. 28,33,34 In other words, besides the hydrophobic region of PLGA being efficient in hydrophobic drug encapsulation, the hydration on the hydrophilic PEG corona can further prevent nanocarriers from aspecific protein adsorption and reticuloendothelial system clearance for prolonging blood circulation in vivo.…”

“…[28][29][30][31][32] Additionally, polyethylene glycolated (PEGylated) PLGA nanoparticles have been developed to improve bioavailability and antitumor targeting further. 28,33,34 In other words, besides the hydrophobic region of PLGA being efficient in hydrophobic drug encapsulation, the hydration on the hydrophilic PEG corona can further prevent nanocarriers from aspecific protein adsorption and reticuloendothelial system clearance for prolonging blood circulation in vivo. [35][36][37][38][39] Therefore, PEGylated PLGA nanoparticles are advanced platforms to enhance targeted antitumor drug delivery.…”