Effect of small interfering RNA delivery using galactosylated chitosan-graft-polyethylenimine on MRE11 gene expression in drug-resistant hepatocellular carcinoma BEL7402/5-Fu cells

doi:10.3969/j.issn.2095-4344.2017.06.020

Abstract

Abstract:

BACKGROUND: Chitosan is a kind of common non-viral vector characterized by good biocompatibility, biodegradability and low toxicity. However, it usually needs structural modification via derivatization due to its weak gene delivery ability.
OBJECTIVE: To observe the ability of galactosylated chitosan-graft-polyethylenimine (GAL-CHI-g-PEI) delivering small interfering RNA (siRNA) on drug-resistant hepatocellular carcinoma BEL7402/5-Fu and its effect on the expression of meiotic recombination 11 (MRE11).
METHODS: BEL7402/5-Fu cells or human hepatocytes HL-7702 were co-cultured with 0, 10, 20, 40, 80 and 100 μg/mL GAL-CHI-g-PEI or chitosan-PEI for 24 hours, and then the cell viability was detected. The composite transfection particles were prepared by mixing GAL-CHI-g-PEI with siRNA-FAM at the mass ratio of 0, 2, 4, 8, 16 and 32, respectively. The next experiments were performed based on BEL7402/5-Fu cells transfected with the compound transfection particles, and the optimal mass ratio and transfection time were selected according to the efficiency of transfection at 18, 72 and 96 hours afte transfection. At 48 hours after BEL7402/5-Fu cells transfected with compound transfection particles, GAL-CHI-g-PEI and siRNA, respertively, the expressions of MRE11 mRNA and protein were detected in comparison with untransfected cells as controls.
RESULTS AND CONCLUSION: Compared with chitosan-PEI, GAL-CHI-g-PEI demonstrated lower toxicity, and still showed low cytotoxicity when the mass concentration reached 100 μg/mL. The transfection efficiency was over 95% and the cell entry efficiency was extremely high when the mass ratio of GAL-CHI-g-PEI to siRNA-FAM was 4 and 8, respectively, with the optimal transfection effect at 48 hours after transfection. Therefore, the mass ratio in the subsequent experiments was set to 8. The expressions of MRE11 mRNA and protein in the composite transfection group were lower than those in the GAL-CHI-g-PEI, siRNA and control groups (P < 0.05). These results suggest that the effective delivery of siRNA and the silencing of the MRE11 gene in BEL7402/5-Fu cells can be achieved by GAL-CHI-g-PEI.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Chitosan, Transfection, Gene Silencing, Polyethyleneimine, Tissue Engineering

CLC Number:

R318

Liu Yun, Pan Jie, Dong Wei, Hui Jing, Li San-hua, Fan Fang, Zhu Xin-ting. Effect of small interfering RNA delivery using galactosylated chitosan-graft-polyethylenimine on MRE11 gene expression in drug-resistant hepatocellular carcinoma BEL7402/5-Fu cells[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(6): 934-939.

Figures/Tables 7

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