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Impact of Mistletoe Triterpene Acids on the Uptake of Mistletoe Lectin by Cultured Tumor Cells.

Mulsow K, Enzlein T, Delebinski C, Jaeger S, Seifert G, Melzig MF - PLoS ONE (2016)

Bottom Line: It could be shown that the intracellular uptake after 120 minutes amounted to 20% in all cell lines after incubation with viscumTT.The studies further revealed that the uptake in THP-1-, HL-60- and Ewing TC-71-cells was independent of the addition of TT extract.Interestingly, the uptake of ML by 143B-cells could only be measured after addition of triterpenes pointing to resistance to mistletoe lectin.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pharmacy, Department of Pharmaceutical Biology, Freie Universitaet Berlin, Berlin, Germany.

ABSTRACT
Complementary treatment possibilities for the therapy of cancer are increasing in demand due to the severe side effects of the standard cytostatics used in the first-line therapy. A common approach as a complementary treatment is the use of aqueous extracts of Viscum album L. (Santalaceace). The therapeutic activity of these extracts is attributed to Mistletoe lectins which are Ribosome-inactivating proteins type II. Besides these main constituents the extract of Viscum album L. comprises also a mixture of lipophilic ingredients like triterpene acids of the oleanane, lupane and ursane type. However, these constituents are not contained in commercially available aqueous extracts due to their high lipophilicity and insolubility in aqueous extraction media. To understand the impact of the extract ingredients in cancer therapy, the intracellular uptake of the mistletoe lectin I (ML) by cultured tumor cells was investigated in relation to the mistletoe triterpene acids, mainly oleanolic acid. Firstly, these hydrophobic triterpene acids were solubilized using cyclodextrins ("TT" extract). Afterwards, the uptake of either single compounds (isolated ML and the aqueous "viscum" extract) or in combination with the TT extract (ML+TT, viscumTT), was analyzed. The uptake of ML was studied inTHP-1-, HL-60-, 143B- and Ewing TC-71-cells and determined after 30, 60 and 120 minutes by an enzyme linked immunosorbent assay which quantifies the A-chain of the hololectin. It could be shown that the intracellular uptake after 120 minutes amounted to 20% in all cell lines after incubation with viscumTT. The studies further revealed that the uptake in THP-1-, HL-60- and Ewing TC-71-cells was independent of the addition of TT extract. Interestingly, the uptake of ML by 143B-cells could only be measured after addition of triterpenes pointing to resistance to mistletoe lectin.

No MeSH data available.


Related in: MedlinePlus

Cell viability of HL-60-cells.The cells were treated with different ML concentrations (see Tables 2 and 3) for 30, 60 and 120 minutes. The isolated ML, three viscum extract batches and the viscum extract batch 161 in combination with TT 161 extract batch (25 μg/mL and 35 μg/mL OA) were used. The viability was determined with Annexin V-APC and propidium iodide by flow cytometry. The values are expressed as percentages of the untreated control cells. Error bars represent the standard deviation of n = 2 experiments.
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pone.0153825.g001: Cell viability of HL-60-cells.The cells were treated with different ML concentrations (see Tables 2 and 3) for 30, 60 and 120 minutes. The isolated ML, three viscum extract batches and the viscum extract batch 161 in combination with TT 161 extract batch (25 μg/mL and 35 μg/mL OA) were used. The viability was determined with Annexin V-APC and propidium iodide by flow cytometry. The values are expressed as percentages of the untreated control cells. Error bars represent the standard deviation of n = 2 experiments.

Mentions: The PI and Annexin V binding assays displayed no significant differences between treated and untreated cells. Control cells had an average cell viability of 100 ± 6.7% and treated cells of 94.1 ± 10.7%. Fig 1 shows the cell viability for the HL-60-cellline.


Impact of Mistletoe Triterpene Acids on the Uptake of Mistletoe Lectin by Cultured Tumor Cells.

Mulsow K, Enzlein T, Delebinski C, Jaeger S, Seifert G, Melzig MF - PLoS ONE (2016)

Cell viability of HL-60-cells.The cells were treated with different ML concentrations (see Tables 2 and 3) for 30, 60 and 120 minutes. The isolated ML, three viscum extract batches and the viscum extract batch 161 in combination with TT 161 extract batch (25 μg/mL and 35 μg/mL OA) were used. The viability was determined with Annexin V-APC and propidium iodide by flow cytometry. The values are expressed as percentages of the untreated control cells. Error bars represent the standard deviation of n = 2 experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835140&req=5

pone.0153825.g001: Cell viability of HL-60-cells.The cells were treated with different ML concentrations (see Tables 2 and 3) for 30, 60 and 120 minutes. The isolated ML, three viscum extract batches and the viscum extract batch 161 in combination with TT 161 extract batch (25 μg/mL and 35 μg/mL OA) were used. The viability was determined with Annexin V-APC and propidium iodide by flow cytometry. The values are expressed as percentages of the untreated control cells. Error bars represent the standard deviation of n = 2 experiments.
Mentions: The PI and Annexin V binding assays displayed no significant differences between treated and untreated cells. Control cells had an average cell viability of 100 ± 6.7% and treated cells of 94.1 ± 10.7%. Fig 1 shows the cell viability for the HL-60-cellline.

Bottom Line: It could be shown that the intracellular uptake after 120 minutes amounted to 20% in all cell lines after incubation with viscumTT.The studies further revealed that the uptake in THP-1-, HL-60- and Ewing TC-71-cells was independent of the addition of TT extract.Interestingly, the uptake of ML by 143B-cells could only be measured after addition of triterpenes pointing to resistance to mistletoe lectin.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pharmacy, Department of Pharmaceutical Biology, Freie Universitaet Berlin, Berlin, Germany.

ABSTRACT
Complementary treatment possibilities for the therapy of cancer are increasing in demand due to the severe side effects of the standard cytostatics used in the first-line therapy. A common approach as a complementary treatment is the use of aqueous extracts of Viscum album L. (Santalaceace). The therapeutic activity of these extracts is attributed to Mistletoe lectins which are Ribosome-inactivating proteins type II. Besides these main constituents the extract of Viscum album L. comprises also a mixture of lipophilic ingredients like triterpene acids of the oleanane, lupane and ursane type. However, these constituents are not contained in commercially available aqueous extracts due to their high lipophilicity and insolubility in aqueous extraction media. To understand the impact of the extract ingredients in cancer therapy, the intracellular uptake of the mistletoe lectin I (ML) by cultured tumor cells was investigated in relation to the mistletoe triterpene acids, mainly oleanolic acid. Firstly, these hydrophobic triterpene acids were solubilized using cyclodextrins ("TT" extract). Afterwards, the uptake of either single compounds (isolated ML and the aqueous "viscum" extract) or in combination with the TT extract (ML+TT, viscumTT), was analyzed. The uptake of ML was studied inTHP-1-, HL-60-, 143B- and Ewing TC-71-cells and determined after 30, 60 and 120 minutes by an enzyme linked immunosorbent assay which quantifies the A-chain of the hololectin. It could be shown that the intracellular uptake after 120 minutes amounted to 20% in all cell lines after incubation with viscumTT. The studies further revealed that the uptake in THP-1-, HL-60- and Ewing TC-71-cells was independent of the addition of TT extract. Interestingly, the uptake of ML by 143B-cells could only be measured after addition of triterpenes pointing to resistance to mistletoe lectin.

No MeSH data available.


Related in: MedlinePlus