Synthesis, characterization, and anticancer effect of nanoformulated Zataria multiflora Boiss. extract against prostate and breast cancer cells in vitro

This study aimed to investigate the effect of nanoformulated Zataria multiflora Boiss. (Shirazi thyme) extract on inhibiting the growth of PC3 prostate and MDA-MB-231 breast cancer cell lines by regulating autophagy. The hydroalcoholic extract of Zataria multiflora Boiss. (ZM) was prepared by ultrasonic-assisted maceration in 70% ethanol. Blank (Gel NP) and extract-loaded gelatin nanoparticles (ZM.Gel NPs) were synthesized via a two-step desolvation method and characterized in terms of size, surface charge, morphology, chemical structure, loading parameters, and in vitro release kinetics. PC3 and MDA-MB-231 cell lines were treated with various concentrations of the free extract, Gel NPs, and ZM.Gel NPs (with equivalent encapsulated ZM extract) for 48 and 72 h. Cell proliferation was evaluated using the MTT assay, and IC50 values were calculated. The expression of apoptosis- and autophagy-related genes was determined using Real-Time PCR. Gel NPs and ZM.Gel NPs exhibited spherical morphology, with smooth surfaces and suitable physicochemical properties, while providing sustained release of ZM extract over time. ZM.Gel NPs significantly enhanced the antiproliferative potential of the free extract, thereby reducing the IC50 value. ZM.Gel NPs significantly upregulated TP53 and BAX and downregulated BCL2 , while decreasing Beclin-1 , ATG5 , ATG7 , and LC3A expression compared with the free extract. Gel NPs enhanced the antiproliferative and cytotoxic effects of ZM extract, upregulating pro-apoptotic genes and downregulating autophagy-driven genes in prostate and breast cancer cell lines.