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Submitted: 11 Jun 2024
Revision: 12 Jul 2025
Accepted: 20 Aug 2025
ePublished: 16 Sep 2025
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Immunopathol Persa. Inpress.
doi: 10.34172/ipp.2025.40686
  Abstract View: 13

Original

Anti-tumor activity of prepared human alpha-lactalbumin-oleic acid and bovine alpha-lactalbumin-oleic acid complexes on mice bearing AN3 murine mammary adenocarcinoma

Omar A. Mahmoud 1* ORCID logo, Shahlaa M. Salih 2 ORCID logo, Zahraa K. Zedan 2 ORCID logo

1 Department of Molecular Genetics and DNA Fingerprint Forensic DNA Center for Research and Training, AL-Nahrain University, Jadriya, Baghdad, Iraq
2 Department of Molecular and Medical Biotechnology, College of Biotechnology, Jadriya, Baghdad, Iraq
*Corresponding Author: Omar A. Mahmoud, Email: omar.abd206@nah-rainuniv.edu.iq

Abstract

Introduction: Current medical issues include the difficulty of selectively targeting and diminishing cancer cells without impacting normal cells, as seen by traditional therapies like chemotherapy and radiation therapy. Recent research has identified proteins with anticancer capabilities, notably alpha-lactalbumin (α-LA), a major protein in human and bovine milk. α-LA, especially in conjunction with oleic acid, has shown specific cytotoxicity towards cancer cells. This research examines the anticancer properties of HAMLET (human alpha-lactalbumin made lethal to tumor cells) and BAMLET (bovine alpha-lactalbumin made lethal to tumor cells) complexes on AN3 murine mammary adenocarcinoma.

Objectives: This study seeks to evaluate the antitumor effect of human α-LA-oleic acid and bovine α-LA-oleic acid complexes against AN3 murine mammary cancer in mice.

Materials and Methods: In this in vivo experimental study, human breast milk was collected from mothers aged 24-28 years in Baghdad, and cow milk was purchased locally. Additionally, α-LA was purified using high-performance liquid chromatography (HPLC), with human α-LA at 1.2597 ppm and bovine α-LA at 1.8352 ppm. Complexes of α-LA with oleic acid were prepared, heated, sonicated, and pH-adjusted. Mice bearing AN3 mammary adenocarcinoma were injected subcutaneously with HAMLET or BAMLET and monitored. Leukocyte counts, interferon-γ levels, and histological analysis of tumors were conducted to assess immune responses and tumor changes.

Results: The HPLC study verified the presence of α-LA in both human and bovine specimens. Mice treated with HAMLET and BAMLET showed significant increases in total leukocyte, lymphocyte, neutrophil, and monocyte counts compared to controls. HAMLET-treated mice exhibited a total leukocyte count of 10920.0±71.14 cells/mL blood, and BAMLET-treated mice showed 10324.0±205.05 cells/mL blood, both significantly higher than the control group’s 9556.6±595.4 cells/mL blood. Interferon-γ levels were notably elevated in both treatment groups, Histopathological analysis revealed pronounced necrosis and increased inflammatory cells in treated tumors.

Conclusion: Both HAMLET and BAMLET complexes exhibited significant antitumor effects on mice bearing AN3 murine mammary adenocarcinoma, enhancing innate and adaptive immune responses and inducing high levels of interferon-γ. These findings suggest potential therapeutic applications for α-LA-oleic acid complexes in cancer treatment.


Citation: Mahmoud OA, Salih SM, Zedan ZK. Anti-tumor activity of prepared human alpha-lactalbumin-oleic acid and bovine alpha-lactalbumin-oleic acid complexes on mice bearing AN3 murine mammary adenocarcinoma. Immunopathol Persa. 2025;x(x):e40686. DOI:10.34172/ipp.2025.40686.
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