Asialoglycoprotein receptor-magnetic dual targeting nanoparticles for delivery of RASSF1A to hepatocellular carcinoma

Abstract

We developed a nanovector with double targeting properties for efficiently delivering the tumor suppressor gene RASSF1A specifically into hepatocellular carcinoma (HCC) cells by preparing galactosylated-carboxymethyl chitosan-magnetic iron oxide nanoparticles (Gal-CMCS-Fe3O4-NPs). After conjugating galactose and CMCS to the surface of Fe3O4-NPs, we observed that Gal-CMCS-Fe3O4-NPs were round with a relatively stable zeta potential of +6.5 mV and an mean hydrodynamic size of 40.1 ± 5.3 nm. Gal-CMCS-Fe3O4-NPs had strong DNA condensing power in pH 7 solution and were largely nontoxic. In vitro experiments demonstrated that Gal-CMCS-Fe3O4-NPs were highly selective for HCC cells and liver cells. In vivo experiments showed the specific accumulation of Gal-CMCS-Fe3O4-NPs in HCC tissue, especially with the aid of an external magnetic field. Nude mice with orthotopically transplanted HCC received an intravenous injection of the Gal-CMCS-Fe3O4-NPs/pcDNA3.1(+)RASSF1A compound and intraperitoneal injection of mitomycin and had an external magnetic field applied to the tumor area. These mice had the smallest tumors, largest percentage of TUNEL-positive cells, and highest caspase-3 expression levels in tumor tissue compared to other groups of treated mice. These results suggest the potential application of Gal-CMCS-Fe3O4-NPs for RASSF1A gene delivery for the treatment of HCC.

Ligand-based targeted therapy: a novel strategy for hepatocellular carcinoma

“…In aqueous solution, the glucose group becomes the chain structure, which can react with amino group. 51,71,72 The Gal group yet maintains a stable ring structure. In addition, some research studies indicated that only a small amount of NPs without galactosamine was internalized into hepatoma cells, whereas the galactosamine-modified NPs lead to high targeting to hepatic tumor and facilitate their cellular uptake.…”

N‐Acetylgalactosamine‐Targeted Delivery of Dendrimer‐Doxorubicin Conjugates Influences Doxorubicin Cytotoxicity and Metabolic Profile in Hepatic Cancer Cells

“…We measured the extent of hemolysis induced by P1 and P2 conjugates in the presence of freshly isolated red blood cells (RBCs) by quantifying the amount of released hemoglobin from ruptured RBCs after a 1 h incubation at 37 °C, and compared this behavior to naked, nonPEGylated G5‐(NH 2 ) 128 dendrimers ( Figure , Panel A). Results are presented as a percentage of hemolysis caused by distilled (DI) water, which is considered to cause 100% hemolysis through osmotic swelling and rupture of RBCs . Unmodified G5‐(NH 2 ) 128 dendrimers exhibited complete hemolysis (98.7 ± 3.1%), which can be attributed to membrane destabilization caused by the cationic quaternary ammonium ions that develop at the amine‐terminated surfaces of PAMAM dendrimers .…”

“…Finally, 200 µL of the supernatant was collected and added to 96‐well plates, and the excitation of hemoglobin was measured by UV (λ ex = 541 nm). The raw data was normalized to PBS values (which is nonhemolytic due to its buffering capacity) and presented as a percentage of hemolysis caused by DI water (which causes hemolysis through osmotic swelling and rupture of RBCs). Results are presented as the mean of three replicates ± SEM.…”

Liver Cancer: Current and Future Trends Using Biomaterials

“…Magnetic iron NPs have the ability to be controlled by superparamagnetism, allowing the regulation of their movement and concentration using an external magnetic field [71]. Xue et al utilized galactosylated-carboxymethyl chitosan-magnetic iron oxide NPs (Gal-CMCS-Fe 3 O 4 -NPs) to deliver the tumor suppressor gene RASSF1A [72]. They were able to improve the transfection efficiency of the particles by controlling the direction of travel of the super paramagnetic iron oxide NPs.…”