Lipids as Tumoricidal Components of Human α-Lactalbumin Made Lethal to Tumor Cells (HAMLET)

Ho, James C. S.; Storm, Petter; Rydström, Anna; Bowen, Ben; Alsin, Fredrik; Sullivan, Louise M.; Ambite, Inès; Mok, K. Hun; Northen, Trent; Svanborg, Catharina

doi:10.1074/jbc.m113.468405

Cited by 21 publications

(14 citation statements)

References 59 publications

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“…While we have no definite explanation for the discrepancy, poor solubility of OA, its tendency to stick to the walls of plastic tubes under the conditions of the study and consequent low availability [17] for interacting with the cells apparently necessitated high concentrations of the fatty acid to induce hemolysis. Oleate, that forms HAMLET-like complexes [41] and its relevant constituent [42], was however more hemolytic than OA and moderately more hemolytic than even α-LA associated OA (Figure 2B). BAMLET prepared by the procedure of Kamijima et al [20], when subjected to crosslinking with 0.01% glutaraldehyde resulted in the formation of adducts that moved in SDS-PAGE as polypeptides ranging from 50 kDa to those that were too large to enter the gel (Figure S2) as also reported by Spolaore et al [19].…”

Section: Resultsmentioning

confidence: 99%

“…During their investigation of HAMLET-induced ion channels in A549 lung carcinoma cells, Storm et al [43] observed that the increase in free intracellular calcium was not restricted by inclusion of EGTA in the medium but by the inhibition of INsP3-gated ER Ca 2+ channel with U73122, implying that it originated mainly from intracellular stores. In another recent study [42] it was however shown that HAMLET and oleate trigger overlapping but fundamentally different responses on the ion channels of the lung carcinoma and Jurkat cells.…”

Section: Resultsmentioning

confidence: 99%

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Oleic Acid May Be the Key Contributor in the BAMLET-Induced Erythrocyte Hemolysis and Tumoricidal Action

et al. 2013

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A chance discovery of the tumoricidal action of a human milk fraction led to the characterization of the active component as oleic acid complex of the α-lactalbumin, which was given the acronym HAMLET. We report in this study that the oleic acid complex of bovine α-lactalbumin (BAMLET) is hemolytic to human erythrocytes as well as to those derived from some other mammals. Indirect immunofluorescence analysis suggested binding of BAMLET to erythrocytes prior to induction of hemolysis. Free OA was hemolytic albeit at higher concentrations, while sodium oleate caused hemolysis at far lower concentrations. Amiloride and BaCl2 offered protection against BAMLET-induced hemolysis suggesting the involvement of a cation leak channel in the process. BAMLET coupled to CNBr-activated Sepharose was not only hemolytic but also tumoricidal to Jurkat and MCF-7 cells in culture. The Sepharose-linked preparation was however not toxic to non-cancerous peritoneal macrophages and primary adipocytes. The tumoricidal action was studied using the MTT-assay while apoptosis induction measured by the annexin V-propidium iodide assay. Repeated incubation of the immobilized BAMLET with erythrocytes depleted oleic acid and decreased the hemolytic activity of the complex. Incubation of MCF-7 and Jurkat cells with OA, soluble or immobilized BAMLET resulted in increase in the uptake of Lyso Tracker Red and Nile red by the cells. The data presented support the contention that oleic acid plays the key role, both in BAMLET-induced hemolysis and tumoricidal action.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Oleic Acid May Be the Key Contributor in the BAMLET-Induced Erythrocyte Hemolysis and Tumoricidal Action

et al. 2013

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“…Typically, some further restrictions are placed on the culture chambers used, as normal cell culture at these time scales requires regular change of the cell culture medium. Consequently, long term studies have used various kinds of perfusion chambers to be able to replace the medium without disturbing the imaging view [30,32,39,43,49,55,61]. Several studies have also used larger culture chambers with large enough volume to avoid changing the medium entirely, such as by using a T25 cell culture flask (Table 1).…”

Section: Choice Of Cell Culture and Imaging Vesselmentioning

confidence: 99%

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

2018

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Abstract:To understand complex biological processes, scientists must gain insight into the function of individual living cells. In contrast to the imaging of fixed cells, where a single snapshot of the cell's life is retrieved, live-cell imaging allows investigation of the dynamic processes underlying the function and morphology of cells. Label-free imaging of living cells is advantageous since it is used without fluorescent probes and maintains an appropriate environment for cellular behavior, otherwise leading to phototoxicity and photo bleaching. Quantitative phase imaging (QPI) is an ideal method for studying live cell dynamics by providing data from noninvasive monitoring over arbitrary time scales. The effect of drugs on migration, proliferation, and apoptosis of cancer cells are emerging fields suitable for QPI analysis. In this review, we provide a current insight into QPI applied to cancer research.

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“…As monitored by DH, 15 min of incubation, with 35 μM of HAMLET, was enough for a reduction in cell area and increase in thickness, with evidence of membrane blebbing in human lung carcinoma A549 cells [26]. After 60 min, the cells became even smaller in area and thicker [30]. Puthia et al examined how HAMLET affects β-catenin and Wnt-signaling in the treated human colon cancer cell line DLD1.…”

Section: Cell Death Of Adherent Cellsmentioning

confidence: 99%

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

El-Schich¹,

Kamlund²,

Janicke³

et al. 2017

Holographic Materials and Optical Systems

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Digital holographic (DH) microscopy is a digital high-resolution holographic imaging technique with the capacity of quantification of cellular conditions without any staining or labeling of cells. The unique measurable parameters are the cell number, cell area, thickness, and volume, which can be coupled to proliferation, migration, cell cycle analysis, viability, and cell death. The technique is cell friendly, fast and simple to use and has unique imaging capabilities for time-lapse investigations on both the single cell and the cell-population levels. The interest for analyzing specifically cell volume changes with DH microscopy, resulting from cytotoxic treatments, drug response, or apoptosis events has recently increased in popularity. We and others have used DH microscopy showing that the technique has the sensitivity to distinguish between different cells and treatments. Recently, DH microscopy has been used for cellular diagnosis in the clinic, providing support for using the concept of DH, e.g., screening of malaria infection of red blood cells (RBC), cervix cancer screening, and sperm quality. Because of its quick and label-free sample handling, DH microscopy will be an important tool in the future for personalized medicine investigations, determining the optimal therapeutic concentration for both different cancer types and individual treatments.

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Lipids as Tumoricidal Components of Human α-Lactalbumin Made Lethal to Tumor Cells (HAMLET)

Cited by 21 publications

References 59 publications

Oleic Acid May Be the Key Contributor in the BAMLET-Induced Erythrocyte Hemolysis and Tumoricidal Action

Oleic Acid May Be the Key Contributor in the BAMLET-Induced Erythrocyte Hemolysis and Tumoricidal Action

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

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