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<p>Dietary chemoprevention offers a low-toxicity strategy to complement traditional cancer therapies. Bovine casein, the primary milk protein fraction, can produce bioactive peptides with reported anticancer properties. However, the influence of cattle genetic lineage on casein composition and activity remains insufficiently understood. This study compared the characteristics of casein-derived native and hybrid cows, evaluated their cytotoxic and apoptotic effects on human colorectal cancer HT-29 cells, and assessed their biocompatibility with Vero cells. Gir milk showed higher protein (10.7 vs. 1.6&#xa0;mg/mL) and carbohydrate (2.4 vs. 1.6&#xa0;mg/mL) contents than Jersey milk, while fat levels differed (4.34 vs. 5.44%), with total solids being similar (~ 16.6–16.8%). Native-PAGE revealed a major casein band along with minor bands in Gir, whereas SDS-PAGE identified two prominent bands (~ 35–25&#xa0;kDa for αs1- and β-casein) and additional minor bands. Chromatographic analysis and MALDI-TOF/MS displayed a single prominent peak in Gir and three in Jersey, with Gir exhibiting higher m/z features (up to ~ 2020.985) and Jersey exhibiting lower m/z features (~ 719.638). Both chromatographic and mass spectrometry analyses indicated peptide profiles, with Gir having higher-mass features and Jersey having lower-mass features. Gir casein demonstrated poor biocompatibility, as evidenced by a dose-dependent decrease in Vero cell viability and an approximately 93.8% loss of viability in Jersey cells. Both caseins showed moderate, dose-dependent cytotoxicity toward HT-29 cells (67.50% for Gir and 75.80% for Jersey at 50&#xa0;µg/mL; 44.80% and 50.09% at 350&#xa0;µg/mL), with morphological apoptosis confirmed by PI staining. Variations in casein and peptide composition among breeds influence their safety and efficacy profiles, highlighting their potential as natural supplements for colorectal cancer; further research is needed to clarify their molecular mechanisms and in vivo effects.</p>

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Casein protein from Gir and Jersey cow milk: biochemical profiling, MALDI-TOF/MS peptide fingerprinting, and cytotoxic effects on HT-29 cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月7日

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  • Sivasamy Sivaranjani,
  • Veerapandiyan Ramabhai,
  • Arumugam Sarumathi,
  • Arockiasamy Anusuya,
  • Rengarajan Murugesan

摘要

Dietary chemoprevention offers a low-toxicity strategy to complement traditional cancer therapies. Bovine casein, the primary milk protein fraction, can produce bioactive peptides with reported anticancer properties. However, the influence of cattle genetic lineage on casein composition and activity remains insufficiently understood. This study compared the characteristics of casein-derived native and hybrid cows, evaluated their cytotoxic and apoptotic effects on human colorectal cancer HT-29 cells, and assessed their biocompatibility with Vero cells. Gir milk showed higher protein (10.7 vs. 1.6 mg/mL) and carbohydrate (2.4 vs. 1.6 mg/mL) contents than Jersey milk, while fat levels differed (4.34 vs. 5.44%), with total solids being similar (~ 16.6–16.8%). Native-PAGE revealed a major casein band along with minor bands in Gir, whereas SDS-PAGE identified two prominent bands (~ 35–25 kDa for αs1- and β-casein) and additional minor bands. Chromatographic analysis and MALDI-TOF/MS displayed a single prominent peak in Gir and three in Jersey, with Gir exhibiting higher m/z features (up to ~ 2020.985) and Jersey exhibiting lower m/z features (~ 719.638). Both chromatographic and mass spectrometry analyses indicated peptide profiles, with Gir having higher-mass features and Jersey having lower-mass features. Gir casein demonstrated poor biocompatibility, as evidenced by a dose-dependent decrease in Vero cell viability and an approximately 93.8% loss of viability in Jersey cells. Both caseins showed moderate, dose-dependent cytotoxicity toward HT-29 cells (67.50% for Gir and 75.80% for Jersey at 50 µg/mL; 44.80% and 50.09% at 350 µg/mL), with morphological apoptosis confirmed by PI staining. Variations in casein and peptide composition among breeds influence their safety and efficacy profiles, highlighting their potential as natural supplements for colorectal cancer; further research is needed to clarify their molecular mechanisms and in vivo effects.