Lady’s-mantle (Alchemilla vulgaris) and Cancer (anticancer research): evidence and sources

Ethnomedicinal records have long mentioned the historical usage of Alchemilla vulgaris L. in folk medicine, particularly for the treatment of gynecological issues. Building on this ethnomedicinal knowledge regarding female illnesses, the aim of this research was to evaluate the impact of ethanolic extract of A. vulgaris on mouse breast cancer cells (4T1) in vitro and in vivo, in addition to its effect on the immune compartment in the tumor microenvironment. Behind viability decrease of 4T1 cells induced by treatment with A. vulgaris extract was strong inhibition of cell proliferation accompanied by caspase-dependent apoptosis and autophagic cell death. Observed changes in 4T1 cell culture after treatment were well orchestrated and led to a reduction in metastatic potential through weakened adhesion, invasion, migration, and colony-forming abilities in vitro. Enhanced intracellular production of reactive oxygen and nitrogen species promoted by the treatment might interfere with all the observed effects. Apart from the direct effect on tumor cells, the A. vulgaris extract significantly reduced tumor growth in the solid orthotropic mammary carcinoma model through restitution of efficient local and systemic immune response reflected in enhanced antigen-presenting potential of dendritic cells (DCs) as well as the extent and activity of effector T cells.

Despite the proven biological activity of the aerial part extract of Alchemilla vulgaris , scarce information is available about the activity of the root extract. This encouraged us to initiate the current investigation to study the cytotoxic activity of A. vulgaris methanolic root extract against various cancer cell lines in vitro, along with its antimicrobial activity and phytochemical screening. MTT assay was applied to test the cytotoxic effect against the prostate (PC-3), breast (MCF-7) and colorectal adenocarcinoma (Caco-2), together with normal Vero cells. Flow cytometry was employed to assess cell cycle arrest and apoptosis vs. necrosis in PC-3 cells. The expression of apoptosis-related genes ( BAX , BCL2 and P53 ) was quantified by qRT-PCR analysis. The obtained results showed strong antiproliferative activity on the three cancer cell lines and the normal Vero cells in a dose-dependent manner. A high selectivity index (SI) was recorded against the three cell lines with PC-3 cells showing the highest SI and the lowest IC 50 . This effect was associated with cell cycle arrest at G1 phase and induction of total apoptosis at 27.18% being mainly early apoptosis. Apoptosis induction was related to the upregulation of the proapoptotic genes P53 and BAX and the downregulation of the antiapoptotic gene BCL2 . Additionally, the extract demonstrated in vitro antibacterial activity against Agrobacterium tumefaciens , Serratia marcescens and Acinetobacter johnsoni . Additionally, it showed antifungal activity against Rhizoctonia solani , Penicillium italicum and Fusarium oxysporium . Seven phenolic acids and seven flavonoids were detected. The predominant phenolic acids were cinnamic and caffeic acids, while hisperdin and querestin were the principal flavonoids. These findings provide clear evidence about the promising proapoptotic effect of A. vulgaris root extract, which contributes to laying the basis for broader and in-depth future investigations.

The current study was designed to evaluate the phytochemical profile and biological properties (antioxidant, enzyme inhibitory and cytotoxic activity) of methanolic, ethanolic, ethyl-acetate and water extracts of Alchemilla vulgaris from Southeast Serbia (central Balkan) which was traditionally used to alleviate and treat many diseases. Bioactive compounds were characterized by LC-MS/MS technique and biological properties were evaluated using antioxidant, enzyme inhibitory and cytotoxic assays. Twenty-six phenolic compounds were quantified in methanolic, ethanolic, ethyl acetate and water extracts, whereas the highest yields were found in ethyl-acetate (EA) extract (gallic acid, caffeic acid, catchin, quercetin). This extract has also shown the greatest antioxidant, anticancer and inhibitory enzyme activities which were demonstrated for the first time in this study. The obtained results indicated that Alchemilla vulgaris from South Serbia possesses high potential for pharmaceutical applications.

Alchemilla vulgaris L. (lady's mantle) was used for centuries in Europe and Balkan countries for treatments of numerous conditions and diseases of the reproductive system, yet some of the biological activities of lady's mantle have been poorly studied and neglected. The present study aimed to estimate the potential of A. vulgaris ethanolic extract from Southeast Serbia to prevent and suppress tumor development in vitro, validated by antioxidant, genoprotective, and cytotoxic properties. A total of 45 compounds were detected by UHPLC-HRMS analysis in A. vulgaris ethanolic extract. Measurement of antioxidant activity revealed the significant potential of the tested extract to scavenge free radicals. In addition, the analysis of micronuclei showed an in vitro protective effect on chromosome aberrations in peripheral human lymphocytes. A. vulgaris extract strongly suppressed the growth of human cell lines derived from different types of tumors (MCF-7, A375, A549, and HCT116). The observed antitumor effect is realized through the blockade of cell division, caspase-dependent apoptosis, and autophagic cell death. Our study has shown that Alchemilla vulgaris L. is a valuable source of bioactive compounds able to protect the subcellular structure from damage, thus preventing tumorigenesis as well as suppressing tumor cell growth.