Influence of fetal bovine serum on cytotoxic and genotoxic effects of lectins in MCF‐7 cells

Faheina–Martins, Gláucia V.; Silveira, Alethéia Lacerda da; Ramos, Márcio V.; Marques‐Santos, Luis Fernando; Araújo, Demétrius Antônio Machado de

doi:10.1002/jbt.20388

Cited by 27 publications

(7 citation statements)

References 32 publications

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“…We also tested the ApoE mimetic peptide under low bovine fetal serum concentrations with glutamine-free medium (1%), a stress condition that significantly reduces cell viability [17]. Triplicate flow cytometry analyses demonstrated, as expected, a significant increase in apoptosis in IEC-6 cells 24 h post-5-FU challenge.…”

Section: Resultsmentioning

confidence: 99%

Apolipoprotein E COG 133 mimetic peptide improves 5-fluorouracil-induced intestinal mucositis

Azevedo

Oliveira

et al. 2012

BMC Gastroenterol

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BackgroundIntestinal mucositis is one of the major troublesome side effects of anticancer chemotherapy leading to poor patient compliance. In this study we addressed the role of the novel apolipoprotein E (ApoE) COG 133 mimetic peptide in 5-fluorouracil (5-FU)-challenged Swiss mice and IEC-6 cell monolayers. Experiments were also conducted in C57BL6J ApoE knock-out mice to assess the effects of apoE peptide treatment.MethodsExperimental groups were as follows: unchallenged controls, 5-FU-challenged mice (450 mg/kg, i.p) with or without the ApoE peptide (0.3, 1, and 3 μM, given twice daily i.p. for 4 days). Mice were sacrificed 3 days after 5-FU challenge. Proximal small intestinal samples were harvested for molecular biology and histological processing. We conducted ELISA assays and RT-PCR to target IL-1β, TNF-α, IL-10, iNOS, and myeloperoxidase (MPO) to assess intestinal inflammation. Cell death and NF-κB assays were also conducted in apoE knock-out mice. In our in vitro models, IEC-6 cells were exposed to 1 mM of 5-FU in glutamine free media with or without the ApoE peptide (0.02, 0.2, 2, 5, 10, and 20 μM). We investigated IEC-6 cell proliferation and migration, 24 h after the 5-FU challenge. Additionally, apoptotic IEC-6 cells were measured by Tunel and flow cytometry. Equimolar doses of the ApoA-I (D4-F) peptide were also used in some experiments for comparative studies.ResultsVillus blunting and heavy inflammatory infiltrates were seen in the 5-FU-challenged group, findings that were partially ameliorated by the ApoE peptide. We found increased intestinal MPO and pro-inflammatory IL-1β and TNF-α levels, and TNF-α and iNOS transcripts, and reduction of IL-10 following 5-FU treatment, each of which were partially abrogated by the peptide. Improvements were also found in IEC-6 cell apoptosis and migration following ApoE and D-4F treatment.ConclusionAltogether, these findings suggest that the novel ApoE COG 133 mimetic peptide can reduce 5-FU-induced intestinal changes and potentially benefit mucositis.

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Section: Resultsmentioning

confidence: 99%

Apolipoprotein E COG 133 mimetic peptide improves 5-fluorouracil-induced intestinal mucositis

Azevedo

Oliveira

et al. 2012

BMC Gastroenterol

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“…Apoptosis is main mechanism of cell death caused by lectins as reported in literature [28,[14][15] . An important marker of cell death is the disruption of mitochondrial membrane accessed by MTT assay.…”

Section: Resultsmentioning

confidence: 99%

Mechanistic Studies of in vitroAnti-Proliferative Potential of Arisaena intermedium Lectin

Gautam¹,

Kaur²,

Thakur³

et al. 2016

IJLSSR

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ABSTRACT-Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti-proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry's method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.

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“…The most potent activity was observed for the melanoma cell line LOX IMVI (> 95% inhibition). It should be noted that the presence of serum in cell culture media can significantly decrease the cytotoxic effects of lectins, as reported previously , and studies on lectins are often carried out with less than 1% serum. The activity assays used in this work were performed under more stringent conditions (5% serum), showing that inhibitory properties of BfL are robust.…”

Section: Discussionmentioning

confidence: 99%

Structural analysis and unique molecular recognition properties of a Bauhinia forficata lectin that inhibits cancer cell growth

et al. 2017

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Lectins have been used at length for basic research and clinical applications. New insights into the molecular recognition properties enhance our basic understanding of carbohydrate-protein interactions and aid in the design/development of new lectins. In this study, we used a combination of cell based assays, glycan microarrays, and X-ray crystallography to evaluate the structure and function of the recombinant Bauhinia forficata lectin (BfL). The lectin was shown to be cytostatic for several cancer cell lines included in the NCI-60 panel; in particular, it inhibited growth of melanoma cancer cells (LOX IMVI) by over 95%. BfL is dimeric in solution and highly specific for binding of oligosaccharides and glycopeptides with terminal N-acetylgalactosamine (GalNAc). BfL was found to have especially strong binding (apparent Kd = 0.5-1.0 nM) to the tumor associated Tn antigen. High-resolution crystal structures were determined for the ligand-free lectin, as well as for its complexes with three Tn glycopeptides, globotetraose, and the blood group A antigen. Extensive analysis of the eight crystal structures and comparison to structures of related lectins revealed several unique features of GalNAc recognition. Of special note, the carboxylate group of Glu126, lining the glycan-binding pocket, forms H-bonds with both the N-acetyl of GalNAc and the peptide amido group of Tn antigens. Stabilization provided by Glu126 is described here for the first time for any GalNAc-specific lectin. Taken together, the results provide new insights into the molecular recognition of carbohydrates and provide a structural understanding that will enable rational engineering of BfL for a variety of applications.

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Influence of fetal bovine serum on cytotoxic and genotoxic effects of lectins in MCF‐7 cells

Cited by 27 publications

References 32 publications

Apolipoprotein E COG 133 mimetic peptide improves 5-fluorouracil-induced intestinal mucositis

Apolipoprotein E COG 133 mimetic peptide improves 5-fluorouracil-induced intestinal mucositis

Mechanistic Studies of in vitroAnti-Proliferative Potential of Arisaena intermedium Lectin

Structural analysis and unique molecular recognition properties of a Bauhinia forficata lectin that inhibits cancer cell growth

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