Effects of targeted nano-delivery systems combined with hTERT-siRNA and Bmi-1-siRNA on MCF-7 cells.
The aim of this study was to evaluate the efficiency of a targeted siRNA nano-delivery system to silence the expression of Bmi-1 and hTERT, and to verify the toxicity of this delivery system in MCF-7 breast cancer cells. The most effective Bmi-1 siRNA and hTERT siRNA sequences were selected using RT-PCR and Western blotting. The polyethyleneimine (PEI)/siRNA nano-condensate was synthesized using PEI and modified using an NGR peptide fragment for targeting to tumor cells. The vector morphology, particle size and zeta potential were observed using an atomic force microscope and a laser particle size analyzer. The MCF-7 breast cancer cell line was transfected with the vector, and cytotoxicity was tested by MTT assays. The transfection efficiency was evaluated by qRT-PCR and Western blotting. Changes in gene expression and apoptosis rate were measured by flow cytometry. The size of LPN carrier and the condensate particle was between 100 and 200 nm and the potentials were close to neutral. There was maximum transfection efficiency and no significant increase in toxicity at 15 pmol/L. Bmi-1 and hTERT expression decreased, but the inhibition rate increased in the hTERT siRNA group, the hTERT+Bmi-1 siRNA group and the hTERT+Bmi-1 siRNA group compared with the scrambled siRNA group and the control group. Moreover, the hTERT+Bmi-1 siRNA group had the highest level of gene silencing. The complex, composed of Lipo, PEI and siRNA, is low toxicity and efficient transfection vectors. The expression level of Bmi-1 and hTERT was decreased by the gene silencing of either Bmi-1 or hTERT, but the effects were more significant when both were silenced simultaneously.
Department of Pathology, Basic Medical College, Henan Key Laboratory for Tumor Pathology, Zhengzhou University Henan Province, China.
Lei
L
Liu
Department of Pathology, Basic Medical College, Henan Key Laboratory for Tumor Pathology, Zhengzhou University Henan Province, China.
Huixiang
H
Li
Department of Pathology, Basic Medical College, Henan Key Laboratory for Tumor Pathology, Zhengzhou University Henan Province, China.
Min
M
Zhang
Department of Pathology, Basic Medical College, Henan Key Laboratory for Tumor Pathology, Zhengzhou University Henan Province, China.
Xinquan
X
Lv
D017209:Apoptosis; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D049109:Cell Proliferation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D061986:MCF-7 Cells; D050997:Nanomedicine; D053758:Nanoparticles; D009842:Oligopeptides / Q000378:metabolism / Q000633:toxicity; D063150:Polycomb Repressive Complex 1 / Q000235:genetics* / Q000378:metabolism; D011094:Polyethyleneimine / Q000378:metabolism / Q000633:toxicity; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D000067492:RNAi Therapeutics / Q000379:methods*; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D014162:Transfection / Q000379:methods*
C069077:BMI1 protein, human; C526428:NGR peptide; D009842:Oligopeptides; D034741:RNA, Small Interfering; D011094:Polyethyleneimine; D063150:Polycomb Repressive Complex 1; C509186:TERT protein, human; D019098:Telomerase
Bmi-1; Breast cancer; MCF-7; PEI; nano-delivery system; siRNA