RESUMO
Styrene maleic acid copolymers (SMA) form discoidal lipid nanoparticles (lipid nanodisks) that mimic plasma high-density lipoproteins. We have previously prepared and characterized lipid nanodisks composed of SMA and the neutral phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). In the present study, we tested whether the surface charges can alter the physicochemical and biological properties of lipid-SMA discoidal particles. Unlike the case of DMPC alone, addition of saline to the buffer was necessary to induce the formation of lipid-SMA complexes containing either 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) or 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), with formation efficiency being dependent on the concentration of charged lipids. After purification, DMPG- or DMTAP-containing discoidal particles with an approximate size of 10 nm were obtained in a manner similar to DMPC alone. Although DMPG and DMTAP appeared to be similarly incorporated into the lipid nanodisks, the zeta potentials of both particles were comparable. That is, no significant differences were observed in the physicochemical properties between the lipid-SMA nanodisks. Compared to DMPC-SMA nanodisks, the uptake of DMPG or DMTAP-containing discoidal particles by RAW264 cells was increased for both particle types, whereas in MDA-MB-231 cells, only DMTAP-containing discoidal particle uptake was increased. In addition, fluorescence microscopy revealed that lipid-SMA nanodisks are localized adjacent to the plasma membrane of RAW264 cells but in MDA-MB-231 cells they accumulated in the center of the cell. Furthermore, these particles caused cytotoxicity in a cell-type dependent manner, with high toxicity in MDA-MB-231. These results raised the possibility that compositional alterations in lipid-SMA discoidal particles may modulate biological reactions in vivo.
Assuntos
Lipoproteínas/química , Maleatos/química , Maleatos/metabolismo , Poliestirenos/química , Poliestirenos/metabolismo , Membrana Celular/química , Dimiristoilfosfatidilcolina/química , Gotículas Lipídicas/química , Lipoproteínas/metabolismo , Nanopartículas/química , Fosfolipídeos/química , Solubilidade , Estireno/químicaRESUMO
Runt-related transcription factor 1 (RUNX1) is generally considered to function as a tumor suppressor in the development of leukemia, but a growing body of evidence suggests that it has pro-oncogenic properties in acute myeloid leukemia (AML). Here we have demonstrated that the antileukemic effect mediated by RUNX1 depletion is highly dependent on a functional p53-mediated cell death pathway. Increased expression of other RUNX family members, including RUNX2 and RUNX3, compensated for the antitumor effect elicited by RUNX1 silencing, and simultaneous attenuation of all RUNX family members as a cluster led to a much stronger antitumor effect relative to suppression of individual RUNX members. Switching off the RUNX cluster using alkylating agent-conjugated pyrrole-imidazole (PI) polyamides, which were designed to specifically bind to consensus RUNX-binding sequences, was highly effective against AML cells and against several poor-prognosis solid tumors in a xenograft mouse model of AML without notable adverse events. Taken together, these results identify a crucial role for the RUNX cluster in the maintenance and progression of cancer cells and suggest that modulation of the RUNX cluster using the PI polyamide gene-switch technology is a potential strategy to control malignancies.