1 H NMR-Based Metabolomics Survey in Breast Cancer Cell Line Treated by Chimera Alpha - Fetoprotein (AFP) Peptide

INTRODUCTION: This study investigates the anticancer potential of the synthetic 15-mer HM-I peptide (SMAIRVSISERSLYI), derived from a chimeric rat/human α-fetoprotein (AFP). HM-I is proposed as a selective anti-cancer agent with minimal effects on normal cells and specificity for targeting hormone-dependent breast cancer cells. METHODS: Nuclear magnetic resonance (NMR)-based metabolomics, combined with LC/MS and GC/MS spectrometry, was utilized to analyze metabolic alterations induced by HM-I in ZR-75-1 breast cancer cells. Cells were cultured under three conditions: (1) with estradiol (positive control), (2) without estradiol (negative control), and (3) with both estradiol and HM-I peptide (treatment group). Spectral data were processed using H NMR and chemometric methods, including partial least squares-discriminant analysis (PLS-DA). Metabolites were identified via the Human Metabolome Database, and altered metabolic pathways were mapped using bioinformatics tools. RESULTS: Estradiol (10⁸⁹ M) stimulated the proliferation of ER⁷ ZR-75-1 cells, while treatment with HM-I peptide (10⁸⁸ M) significantly inhibited this estrogen-induced proliferation. Metabolic profiling revealed notable alterations primarily in amino acid and carbohydrate metabolism, including pathways related to aminoacyl-tRNA biosynthesis, glycolysis, and biotin synthesis. DISCUSSION: HM-I peptide demonstrates potent inhibitory effects on estradiol-induced breast cancer cell growth, with distinct metabolic alterations. CONCLUSION: In summary, the chimeric peptide HM-I at 10⁸⁸ M concentration effectively inhibits estrogen-induced proliferation in breast cancer cells, likely through modulation of estrogen signaling pathways, without direct cytotoxic effects in the absence of estrogen. These results suggest HMI acts as a selective modulator of hormone-dependent cancer growth. Nevertheless, further quantitative and mechanistic studies are required to fully understand its mode of action. Additionally, in vivo investigations addressing peptide stability, delivery, pharmacokinetics, and safety are crucial for advancing HM-I toward clinical application in estrogen-dependent cancers.