Study on the Improvement of Transfection Efficiency of Antisense Oligonucleotides and Its in Vivo Anti-tumor Effect by Reduction Responsive Micelle Based on Polyethyleneimine
10.13748/j.cnki.issn1007-7693.20231077
- VernacularTitle:基于聚乙烯亚胺的还原响应型胶束提高反义寡核苷酸转染效率及其体内抗肿瘤作用研究
- Author:
Shuang YANG
1
;
Fei PAN
2
;
Beibei HE
2
;
Minfei SHI
2
;
Cuiping HE
2
;
Bin ZHENG
2
Author Information
1. Shanxi Medical University School of Basic Medical Sciences
2. Shanxi Medical University School of Pharmacy, Taiyuan 030001, China
- Publication Type:Journal Article
- Keywords:
polyethyleneimine； reduction responsive ； antisense oligonucleotide；transfection；anti-tumor
- From:
Chinese Journal of Modern Applied Pharmacy
2024;41(9):1159-1167
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE :To synthesize a polymer PEI-ss-PEG2000-DSPE containing disulfide bond and to prepare as cationic micelle(P-ss-PD) based on branched polyethyleneimine(PEI). To investigate the ability of P-ss-PD micelle to reduce cytotoxicity and improve the transfection efficiency of antisense oligonucleotide(ASO) in human breast cancer cell lines, and to study the anti-tumor effect of P-ss-PD micelle in nude mice.
METHODS
PEI-ss-PEG2000-DSPE was synthesized by grafting PEG2000-DSPE onto branched PEI with disulfide bond as a connecting arm. P-ss-PD micelle was prepared by ethanol injection method and P-ss-PD/ASO nanocomplex was obtained by combining P-ss-PD micelle with ASO. The particle size and zeta potential of P-ss-PD/ASO nanocomplex at various mass ratios were determined by laser particle size analyzer. Agarose gel retardation assay was used to investigate the binding degree of P-ss-PD/ASO nanocomplex and determine the optimal mass ratio. At the same time, the reduction responsive ability of P-ss-PD micelle was investigated. The cytotoxicity of P-ss-PD micelle was detected by CCK8 kit. The transfection efficiency of P-ss-PD micelle was investigated by flow cytometry and high content cell imaging analysis system in MDA-MB-231 cells. The anti-tumor effect of P-ss-PD micelle was investigated by tumor-bearing nude mice models.
RESULTS
When the mass ratio was 300∶1, the particle size of P-ss-PD/ASO nanocomplex was the smallest and had a good stability. The average particle size was (58.90 ± 4.08)nm, the average zeta potential was (16.80 ± 1.23)mV, and the morphology was uniform spherical. P-ss-PD/ASO nanocomplex had the reduction responsive ability and could release ASO under highly reductive conditions.In vitro, compared with unmodified branched PEI, the cytotoxicity of P-ss-PD micelle was significantly reduced and the transfection efficiency was significantly increased.In vivo, the tumor growth inhibition rate of P-ss-PD/ASO nanocomplex in tumor-bearing nude mice was more than 50%.
CONCLUSION
The P-ss-PD micelle prepared in this study is a kind of low toxicity and high transfection efficiency non-viral vector, which has the characteristics of reduction responsive releasing, and shows a promising application in ASO drug delivery.
