Design and characterization of pluronic nanomicelles containing retinoic acid and sodium butyrate and evaluating targeted therapeutic potential in MDA-MB-231 breast cancer cell

Abstract

Introduction: Malignant neoplasms are characterized by loss of cellular differentiation and uncontrolled proliferation. Therapeutic differentiation with the aim of restoring endogenous differentiation programs is one of the significant strategies. ATRA causes different types of stem cells and breast cancer cells to differentiate. However, clinical trials of ATRA have shown limited therapeutic success due to the lack of effective delivery systems and, in part, to frequent epigenetic silencing of RARb. Histone deacetylase inhibitors re-express RARb and sensitize cells to treatment. Also, one of the beneficial results of treatment with drug delivery systems is increasing the effectiveness and half-life of the drug, reducing toxicity and the required dose of drugs. Materials and methods: Nanoparticles were formed by forming a thin layer and hydrating it. Interactions with FT-IR and drug loading and release were investigated in acidic and neutral environment. The absorption of nanoparticles and the toxicity effects of nanoparticles on MDA-MB-231, MDA-MB-468 and MCF-7 cell lines were performed. The scratch and spheroid test were performed and the expression of ALDH, CD44, CD24 and RARb genes in all three lines was checked with HA-PF127@ATRA@SB and the apoptosis test was analyzed in three cell lines. Results: The binding of hyaluronic acid to Pluronic was confirmed by 1H-NMR. The size of HA-PF127@ATRA, HA-PF127@SB and HA-PF127@ATRA@SB nanoparticles were 32.02 nm, 47.46 nm and 52.10 nm, respectively. The morphology of the nanoparticles was spherical. FT-IR results indicated physical interaction of drugs with Pluronic micelles. ATRA loading rate was around 96.25% and its release was higher than normal in acidic conditions. The cellular absorption of nanoparticles was higher than the fluorescent substance alone, and the toxicity test showed that MDA-MB-231 was resistant to ATRA and HA nanoparticles HA-PF127@ATRA@SB were able to break the resistance. IC50 of formulated drugs were lower than blank drugs. In the migration test, inhibition of MCF-7 was more than MDA-MB-231, and the size of spheroids decreased the most in the treatment with HA-PF127@ATRA@SB. The results of real time-PCR in the treatment with combination therapy in MDA-MB-231 reduces the characteristics of stemness and in MDA-MB-468 and MCF-7 it causes a relative increase in the characteristics of stemness and malignancy. In the apoptosis test, in MDA-MB-231, a large percentage of the population entered the Late apoptosis phase, and in MDA-MB-468 and MCF-7, most of them are in the Early apoptosis phase. Conclusion: Synthesized nanomicelles have good size and interaction characteristics with drugs and show better performance compared to empty drugs. Also, the adopted therapeutic combination has been able to inhibit MDA-MB-231, which has primary and metastatic characteristics, and abrogate ATRA resistance.