Citation: Jelaˇ ca, S.; Daji´ c-Stevanovi´ c, Z.; Vukovi´ c, N.; Kolašinac, S.; Trendafilova, A.; Nedialkov, P.; Stankovi´ c, M.; Tani´ c, N.; Tani´ c, N.T.; Acovi´ c, A.; et al. Beyond Traditional Use of Alchemilla vulgaris: Genoprotective and Antitumor Activity In Vitro. Molecules 2022, 27, 8113. https://doi.org/10.3390/ molecules27238113 Academic Editors: Irwin Rose Alencar Menezes, Henrique Douglas Melo Coutinho, Almir Gonçalves Wanderley and Jaime Ribeiro-Filho Received: 7 November 2022 Accepted: 19 November 2022 Published: 22 November 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). molecules Article Beyond Traditional Use of Alchemilla vulgaris: Genoprotective and Antitumor Activity In Vitro Sanja Jelaˇ ca 1 , Zora Daji´ c-Stevanovi´ c 2 , Nenad Vukovi´ c 3 , Stefan Kolašinac 2 , Antoaneta Trendafilova 4 , Paraskev Nedialkov 5 , Miroslava Stankovi´ c 6 , Nasta Tani´ c 6 , Nikola T. Tani´ c 1 , Aleksandar Acovi´ c 7 , Sanja Mijatovi´ c 1 and Danijela Maksimovi´ c-Ivani´ c 1, * 1 Department of Immunology, Institute for Biological Research “Siniša Stankovi´ c”—National Institute of the Republic of Serbia, University of Belgrade, Bulevar Despota Stefana 142, 11060 Belgrade, Serbia 2 Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade, Serbia 3 Department of Chemistry, Faculty of Science, University of Kragujevac, Radoja Domanovi´ ca 12, 34000 Kragujevac, Serbia 4 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., bl. 9, 1113 Sofia, Bulgaria 5 Departmentof Pharmacognosy, Faculty of Pharmacy, Medical University of Sofia, Dunav Str. 2, 1000 Sofia, Bulgaria 6 Laboratory for Radiobiology and Molecular Genetics, Institute of Nuclear Sciences—National Institute of the Republic of Serbia, University of Belgrade, Mike Petrovi´ ca Alasa 12–14, 11351 Belgrade, Serbia 7 Department of Dentistry, Faculty of Medical Sciences, University of Kragujevac, Svetozara Markovi´ ca 69, 34000 Kragujevac, Serbia * Correspondence: nelamax@ibiss.bg.ac.rs; Tel.: +381-11-2078452 Abstract: 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. Keywords: Alchemilla vulgaris L.; antitumor action; antioxidative activity; genoprotective effect 1. Introduction Ethnopharmacological data are of crucial importance for the finding of new promising bioactive compounds, as well as for the verification of already accepted herbal drugs. Therefore, it is necessary to preserve the traditional knowledge of medicinal plants in addition to addressing the need for their sustainable collection from the wild. Nowadays, therapeutic approaches have switched from attacking and directly destroying the damaged cells and pathogenic microorganisms towards the activation of self-healing and protective processes upon the initiation of different repair mechanisms of the human body. Such a view has a great impact on scientific research focusing on the bioactivity of natural products [1]. Numerous pathological conditions cannot be fully treated by standard pharmaceutics [2], Molecules 2022, 27, 8113. https://doi.org/10.3390/molecules27238113 https://www.mdpi.com/journal/molecules