Copper or Manganese Supplementation Endows the Peptic Hydrolysate from Bovine Lactoferrin with Enhanced Activity to Human Gastric Cancer AGS Cells
A lactoferrin hydrolysate (LFH) was generated from bovine lactoferrin by pepsin, mixed with Cu2+ and Mn2+ at 0.64–1.28 and 0.28–0.56 mg/g protein, respectively; and then their in vitro effects on human gastric cancer AGS cells were assessed. With incubation times of 24 or 48 h, LFH and its Cu2+/Mn2+ mixtures at 10–30 mg/mL in dose-dependent manner inhibited cell growth; and more, these mixtures showed higher activities than LFH alone. Cell treatments of LFH and the mixtures (25 mg/mL) for 24 h could arrest cell cycle at G0/G1-phase, damage mitochondrial membrane integrity, and induce apoptosis, while the mixtures were also more powerful than LFH to exert these three effects. Higher Cu2+/Mn2+ supplementation level resulted in higher growth inhibition, cell cycle arrest, mitochondrial membrane potential disruption, and apoptosis induction; furthermore, Mn2+ was notable for its higher efficacy than Cu2+ to increase these four effects. Western-blot assay results revealed that four apoptosis-related proteins Bad, Bax, cytochrome c, and p53 were up-regulated, and both caspase-3 and caspase-9 also were cleaved and activated; moreover, two autophagy-related proteins LC3-II and cleaved Beclin-1 were down- and up-regulated, respectively. It is thus concluded that Cu2+ and especially Mn2+ could endow supplemented LFH with increased anti-cancer effects in AGS cells, with two proposed events as enhanced apoptosis induction (via activating apoptosis-related proteins) and autophagy inhibition (via activating autophagy-related proteins).
KeywordsLactoferrin hydrolysate Copper Manganese Gastric cancer cells Apoptosis Autophagy
This work was funded by the Innovative Research Team of Higher Education of Heilongjiang Province (Project No. 2010td11). The authors thank Prof. Li-Ling Yue from Qiqihar Medical University for his kindly help in Western-blot assay, and also thank the anonymous referees for their valuable advice.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no competing interests.
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