视黄酸-伊立替康脂质体制备及其抗甲状腺未分化癌体外评价OA

Objective:To prepare retinoic acid(RA)and irinotecan(IRI)co-loaded liposomes(RA-IRI@Lip)and evaluate their in vitro therapeutic effects against anaplastic thyroid carcinoma(ATC)and their ability to eliminate cancer stem cells(CSCs).Methods:RA-IRI@Lip was obtained through the co-encapsulation of RA and IRI within liposomes,using the thin-film hydration method.The particle size,Zeta potential,encapsulation efficiency,and in vitro release profiles of RA-IRI@Lip were characterized.Hemolysis assay was conducted to evaluate blood compatibility.Cellular uptake of FITC-labeled liposomes was assessed.The in vitro anti-ATC effects were evaluated by cell counting kit-8(CCK-8),calcein-AM/PI staining,and Transwell assay.The synergistic effect of RA and IRI was analyzed using the combination index,while flow cytometry was performed to detect the proportions of aldehyde dehydrogenase-positive cells and CD133-positive cells.Results:The prepared RA-IRI@Lip had an average particle size of(210.3±5.2)nm,a polydispersity index less than 0.3,and Zeta potential of －(13.7±2.3)mV.The encapsulation efficiencies of RA and IRI were(88.1±2.9)%and(86.8±1.6)%,respectively.In vitro release studies showed that RA-IRI@Lip significantly delayed the release of both RA and IRI,with 48-h cumulative release rates of(66.2±3.6)%and(77.7±4.1)%,respectively.Hemolysis assays demonstrated that all tested concentrations of RA-IRI@Lip exhibited hemolysis rates below 3%,indicating good hemocompatibility.Liposomes can be efficiently taken up by ATC cells after FITC labeling.CCK-8 and calcein-AM/PI staining demonstrated that RA-IRI@Lip significantly inhibited the viability of ATC cells and promoted cell death,with an IC50 significantly lower than those of the free IRI group and the free RA+IRI group(both P<0.01).Combination index analysis demonstrated that both the free RA+IRI group and the RA-IRI@Lip group exhibited synergistic effects(combination index<1),with stronger synergism observed in the RA-IRI@Lip group.Transwell assay results demonstrated that RA-IRI@Lip exerted the most significant inhibitory effect on the migration and invasion of ATC cells.Flow cytometry results showed that RA-IRI@Lip significantly decreased the proportions of aldehyde dehydrogenase-positive cells and CD133-positive cells(both P<0.01).Conclusion:RA-IRI@Lip was successfully prepared,achieving synergistic effects of differentiation therapy and chemotherapy of RA and IRI,thereby effectively eliminating ATC cells and CSCs.