Magnetic Nanoparticles Fe₃O₄ synergy with Adriamycin and Arsenic Trioxide Reverse Multidrug Resistance in Lymphoma in Vitro

The aim of this study was to optimize the conditions of combined effect of magnetic nanoparticles Fe₃O₄, adriamycin(ADM) and arsenic trioxide(As₂O₃), search for low toxicity and efficient MDR reversal way and investigate the effect and side reactions of the koinonia in lymphoma cells Raji and Jurket in vitro, which would provide the basis of theory and experiment for in vivo and clinical experiments.

The experimental team regulated quantites of combination which included Fe₃O₄, ADM and As₂O₃ with MTT assay. In a typical experiment, the Raji and Jurket cells were treated with ADM, As₂O₃, ADM and As₂O₃, or Fe₃O₄ synergy with ADM and As₂O₃ for 48 hours, and the cells treated without any drugs were used as control group. Cells apoptosis, and the concentration of ADM within the cells were measured by Flow cytometry (FCM). The distribution of ADM within the cells and cells morphous were observed by fluorescence microscope. The expressions of mRNA and protein of survivin, bcl-2/bax, caspase3, p53 and NF-κB were determined by semi-quantitative reverse transcription PCR (RT-PCR) and Western blot, respectively.

The experimental team selected the synergy quantities of ADM and As₂O₃ at 0. 5mg/L and 1μmol/L or 1. 0mg/L and 2μmol/L cooperated with Fe₃O₄ at 40mg/L while the best synergetic MDR reversal effect came out in both Raji cells and Jurket cells in vitro. The fresh evidence from flow cytometry showed that a higher apoptosis rate could be induced, and a higer concentration of ADM could be detected in lymphoma cells by group of Fe₃O₄ synergy with ADM and As₂O₃ as compared with those by ADM, As₂O₃ or ADM and As₂O₃ in the same concentrations(P<0. 05). The observation from fluorescence microscope presented the distribution of ADM an obvious apoptotic morphous including cell shrinkage, karyopycnosis, nuclear fragmentation, apoptotic body and so on. RT-PCR revealed that the expressions of survivin, bcl-2, and NF-κB mRNA decreased and the expressions of bax, caspase3 and p53 mRNA increased in the group of ADM and As₂O₃ or group of Fe₃O₄ synergy with ADM and As₂O₃ (P<0. 05), but there was no obvious change in other groups(P>0. 05). Western bolt demonstrated that the expressions of survivin, bcl-2, and NF-κB protein were much lower and the expressions of bax, caspase3 and p53 protein were higher in the cells treated with the ADM and As₂O₃ group or Fe₃O₄ synergy with ADM and As₂O₃ group(P<0. 05).

ADM combined As₂O₃ could reverse MDR in lymphoma cells and This synergy with the addition of Fe₃O₄ could enhance the reversal effect. Magnetic nanoparticles(MNPs) Fe₃O₄ copolymerization ADM and As₂O₃ could inhibit ADM eliminated from lymphoma cells and reversing MDR. Because of the ADM retention which could decrease the side effect on the normal tissues and play a persistent reaction to the targeted cells, the effect of inducing apoptosis or regulating gene expressions of Fe₃O₄ synergy with ADM and As₂O₃ group was a little stronger than group of ADM and As₂O₃. Survivin, bcl-2/bax, caspase3, p53 and NF-κB may play an important role in apoptosis induced by Fe₃O₄. More specific apoptosis mechanisms shoud be investigated for the treatment of MNPs in haematopoietic malignancies.

No relevant conflicts of interest to declare.