In-vitro (2D and 3D cultures) and in-vivo cytotoxic properties of Zataria multiflora essential oil (ZEO) emulsion in breast and cervical cancer cells along with the investigation of immunomodulatory potential

Zataria multiflora is an iranian valuable traditional plants, called Avishan Shirazi in Persian language used to reduce inflammation, spasm, pain, and cancer symptoms. Zataria essential oil (ZEO) is one of the essential oils possessing broad biological activities. The aim was to investigate the anticancer effects of ZEO both in-vitro and in-vivo using mouse mammary carcinoma 4T1 cell line and mouse cervical cancer TC1 cell line. The in-vitro effects of ZEO on the proliferation of these cell lines were considered in 2D and 3D culture by MTT assay. In the following, to indicate death mode, fluorescence staining, AnnexinV/PI flowcytometry and caspase-3 activity assay of monolayer cells treated with ZEO was done. In order to evaluate the antitumor activities of ZEO, tumor-bearing BALB/c and C57BL/6 mice were intraperitoneally administered with ZEO and the immunomodulatory effects of ZEO were considered through cytokine assay. Additionally, hematobiochemical factors including aspartate aminotransferase and alanine aminotransferase were investigated to confirm the harmless effects of ZEO. The In-vitro results showed that treatment of cells with ZEO leads to significant inhibition of 4T1 and TC1 cell proliferation and apoptosis in monolayer cell culture (2D) and multicellular spheroids (3D). Based on In-vivo results, ZEO was effective in decreasing the tumor weight compared to the control. Furthermore, ZEO was effective in tilting the balance of cytokines in favor of T helper 1 through the increase in the secretion of TNF-α, IFN-γ, IL-2 and decrease in IL-4. During the treatment with ZEO, hematobiochemical factors of mice did not significantly change. the present study demonstrated that the ZEO has potent antiproliferative, apoptosis-inducing and immune system stimulant properties in breast and cervical cancer. [Display omitted] •ZEO prevents the proliferation of 4T1 and TC1 cells in 2D and 3D cultures.•ZEO induces the apoptosis in 4T1 and TC1 cells.•ZEO reduces tumor growth in 4T1 and TC1 tumor-bearing mice.•ZEO modulates the immune response to reduce tumor growth.