Self-Assembled Fe-LOR-2501 Nanoparticles Loaded Rose Bengal Treat Oral Squamous Cell Carcinoma

Objective: To verify the gene silencing effect and killing effect of Fe-LOR-2501-RB nanoparticles（Fe LORR）by co-delivery of genes and photosensitizer,Rose Bengal（RB）and antisense oligonucleotide LOR-2501 have synergistic effects on oral squamous cells.Methods: First self-assembled prepared by one-step synthesis method of Fe-LOR-2501-RB nanoparticles,through electrostatic adsorption between iron and nucleic acid,ion and nucleic acid in combination of nanostructures,then negatively charged photosensitizer according to certain proportion into the reaction system,and nucleic acid and ionic union,form the photosensitizer and gene delivery system of nanometer.The surface morphology and size of the particles were observed by scanning electron microscopy and projection electron microscopy.Dynamic light scattering experiment to measure particle potential;The absorption peak curve of the nanoparticles was measured by uv spectrophotometer,the photodynamic effect of the nanoparticles was detected by a full-wavelength microplate analyzer,and the biosafety of the nanoparticles and the killing effect of light on oral squamous cell carcinoma CAL-27 were measured by CCK-8.Confocal microscopy was used to determine cell uptake levels and ROS production by flow cytometry.The expression level of R1 gene was detected by Real-time PCR in vitro.Annexin V-FITC/PI apoptosis detection kit was used to detect cell apoptosis.Western Blot assay was used to detect the expression of R1 protein.Results: Scanning electron microscopy（SEM）and transmission electron microscopy（TEM）observed that the synthesized nanoparticles have smooth surface,uniform shape,circular structure and good dispersion.The maximum absorption peak was about 550 nm measured by uv spectrophotometer.The survival rate of cells was lower than 10%when the concentration was 40 m g/ml measured by CCK-8.Confocal microscopy and flow cytometry confirmed that nanoparticles have good delivery effect for gene and photosensitizer.Real-time PCR results showed that Fe-LOR-2501-RB nanoparticles significantly reduced the expression level of R1 gene compared with the control group.Annexin V-FITC/PI apoptosis staining showed that the apoptosis rates of photosensitizer and genome cells were 10.3%,19.2% and 93.3% in the light and photosensitizer groups,respectively.Western Blot analysis showed that the co-delivery gene and photosensitizer group significantly down-regulated the expression of R1 protein.Conclusions: Fe-LOR-2501-RB nanoparticles delivery system can effectively deliver genes and photosensitizers,and achieve good killing effect.Compared with photosensitizers alone,nanoparticles combined with gene therapy can synergistically kill tumors and increase tumor cell apoptosis.