Construction and therapeutic efficacy study of spherical B-cell lymphoblastoma-2 gene antisense oligonucleotide nanoparticles loaded with doxorubicin for enhanced anti-leukemia
Objective To design and self-assemble a novel carrier-free spherical nucleotide nanoparticles for combined treat-ment of leukemia,and study its morphology,physical and chemical properties,drug sustained release behavior and in vitro and invivo efficacy.Methods The 1,3-dilinolenin and B-cell lymphoblastoma-2(Bcl-2)antisense phosphorothioate oligodeoxynucleotide(G3139)were coupled by chemical reaction to form spherical nucleotides with nanocapsule structure by self-assembly,and chemical therapeutic drug daunorubicin(DNR)was encapsulated to construct combined therapeutic spherical nucleotide self-carrier nanomedicine(DBD NP).The intermolecular coupling was detected by agarose gel electrophoresis;particle size,Zeta potential,morphology and stability were determined by dynamic light scattering(DLS)and transmission electron microscopy(TEM);sustained release of DBD nano-drug was detected by enzyme calibration;Sustained release behavior of DBD nanomedicine was detected by enzyme-linked immunosorbent ussay(ELISA).Uptake of DBD NP was observed by laser confocal scanning microscope,and therapeutic effect of DBD NP on leukemia model rats was studied by small animal in vivo imager and flow cytometry.Re-sults DBD NP were spherical nanoparticles with diameter of 140.5 nm,and Zeta potential was(-26.34±2.66)mV.In simulated blood environment,DBD NP exhibit excellent stability within 7-day and sustainably release drug DNR,and cumulative drug release amount reached 76.53％±2.01％on the 5th day.In uptake experiment of LT12 cells,the fluorescence intensity of DNR in DBD NP group was 71.03 × 105,which was about 8 times that of free DNR group,indicated that nanoparticles significantly increased drug uptake of leukemia cells.Animal experiments further confirmed that DBD NP significantly improved therapeutic effect of drugs.The ability to kill leukemia cells was 2-4 times higher than that of free DNR group,and survival time of tumor-bearing rats was prolonged from 23-day to 29-day.Conclusion It is demonstrated that DBD NP could enhance cellular uptake of DNR and G3139 in leukemia cells,thereby impeding the progression of rat leukemia,which is a novel nano-drug with significant potential.
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