AT11-guided liposomes for oral cancer cells: From characterization towards in vitro evaluation

The increasing prevalence of oral cancers, particularly those attributed to human papillomavirus infection, along with the associated drawbacks of conventional therapies, has spurred research into more effective treatment modalities. Liposomes have emerged as a potential strategy to improve the delivery of anticancer molecules to the cells of interest. However, functionalizing these nanoparticles with targeting agents could significantly enhance their selectivity. A prospective approach involves the incorporation of aptamers, which facilitate drug accumulation within cancer cells. Among these, the AT11 aptamer has garnered attention due to its improved toxicity and high affinity to nucleolin, making it a promising targeting moiety. Therefore, we propose to use AT11 for liposomes’ functionalization, to enhance the selectivity of C 8 , a potential anticancer compound, specifically targeting oral cancer cells. Thus, we produced liposomes (empty or C 8 -associated) by ethanol injection method and, then, proceeded with their functionalization with AT11-TEG-Cholesteryl. The resulting liposomes were characterized by dynamic light scattering and hydrodynamic diameters of 130–136 nm were obtained. Additionally, the effect of the produced liposomes on the viability of squamous cell carcinoma of the tongue (UPCI-SCC-154) and nonmalignant (Het1A) cells was determined, by MTT assay. It was demonstrated that the viability of cells treated with empty liposomes was almost unaffected until the 53.6 μg/mL concentration. After treating the cells with C 8 -associated liposomes, both cell lines showed a dose-response effect. Moreover, the AT11-functionalization of the obtained C 8 -associated liposomes enhanced the selectivity of the liposomes towards oral cancer cell line, as observed by the MTT assay (43.2 % in UPCI-SCC-154 versus 79.4 % in Het1A) and confocal microscopy. Furthermore, the anticancer potential of AT11 C 8 -associated liposomes was evaluated in terms of proliferation (ki67 positive cells), cell death (propidium iodide internalization in non-permeabilized cells and caspase-3 activity), migration (cell scratch assay) and invasion. Overall, the produced liposomes decreased tongue cancer cell proliferation (53.6 % versus 99.4 %), migration (from 24.6 % to −3.8 %) and invasion (to 19 %) and induced cell death (11.6-fold increase in propidium iodide positive cells, and a 140 % of caspase-3 activity). These findings suggest that the AT11 C 8 -associated liposomes are promising drug carriers for oral cancer therapy.