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Akacid medical formulation induces apoptosis in myeloid and lymphatic leukemic cell lines in vitro and in vivo.

Neuwirt H, Wabnig E, Feistritzer C, Eder IE, Salvador C, Puhr M, Culig Z, Massoner P, Tiefenthaler M, Steurer M, Konwalinka G - PLoS ONE (2015)

Bottom Line: We have previously shown that AMF exerts antiproliferative effects on a variety of solid tumor cell lines.This effect was found also in G0-arrested cells.Finally, in our animal experiments utilizing male nu/nu Balb/c mice we found a significant growth retardation, which was immunohistochemically associated with a significantly lower number of KI67-positive cells and caspase-3 induction in AMF-treated mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine IV, Medical University of Innsbruck, Innsbruck, Austria.

ABSTRACT
Akacid medical formulation (AMF) is an oligoguanidine that exerts biocidal activity against airborne and surface microorganisms including bacteria, viruses, fungi, and molds, while showing relatively low toxicity to humans. We have previously shown that AMF exerts antiproliferative effects on a variety of solid tumor cell lines. In this study we raised the question whether AMF could also substantially inhibit cell growth or induce apoptosis in cell lines derived from hematologic malignancies such as leukemia or lymphoma. We found that AMF has antiproliferative effects on various hematologic cell lines derived from human leukemia and lymphoma. Additionally, we show that AMF induces apoptosis in leukemia cell lines not only via the extrinsic and intrinsic pathway, but also in a caspase-independent manner. This effect was found also in G0-arrested cells. Finally, in our animal experiments utilizing male nu/nu Balb/c mice we found a significant growth retardation, which was immunohistochemically associated with a significantly lower number of KI67-positive cells and caspase-3 induction in AMF-treated mice.

No MeSH data available.


Related in: MedlinePlus

Caspases are activated after AMF treatment.Capase -8, -9, and -3/-7 activity was measured using a luminometric assay after treatment with various concentrations of AMF for different time periods (4A). n = 3 (number of independent experiments carried out in triplicates). Furthermore Western blot analyses were performed for caspase activation and additionally for cleaved PARP, as a marker for apoptosis induction (4B). n = 3 (number of independent experiments, respresentative blots are shown)
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pone.0117806.g004: Caspases are activated after AMF treatment.Capase -8, -9, and -3/-7 activity was measured using a luminometric assay after treatment with various concentrations of AMF for different time periods (4A). n = 3 (number of independent experiments carried out in triplicates). Furthermore Western blot analyses were performed for caspase activation and additionally for cleaved PARP, as a marker for apoptosis induction (4B). n = 3 (number of independent experiments, respresentative blots are shown)

Mentions: In order to investigate whether the extrinsic or intrinsic apoptosis pathway was involved in AMF-induced apoptosis, we assessed caspase 8, -9 and -3/-7 activity using luminometric assays. As shown in Fig. 4A, AMF induced activity of all tested caspases in a concentration- and time-dependent manner. In particular, caspase 8 was induced already 8 hours after application of 3 μm of AMF in CEM C7H2, whereas in HL-60 higher concentrations and a longer treatment (10 μM for 24 hours) were necessary to yield a similar effect. Analogous results were found for caspase 9 activity. Finally, also caspase 3/-7 activity was induced in both cell lines, with CEM C7H2 being the more sensitive one. These results were corroborated on protein basis utilizing Western blotting for cleaved PARP, caspase -3, -8 and procaspase-9 (Fig. 4B). Unfortunately, we had to stick to an anti-procaspase-9 antibody (due to technical reasons), which when cleaved/activated should be decreased at protein level. However, as anticipated we found a profound decrease of procaspase-9 levels, suggestive for activation, which was found as stated above in experiments for Fig. 4A. Hence, we conclude that AMF induces apoptosis via both the extrinsic and the intrinsic pathway, which has been found to be the case also with various naturally occurring and synthetic molecules [14,15,16].


Akacid medical formulation induces apoptosis in myeloid and lymphatic leukemic cell lines in vitro and in vivo.

Neuwirt H, Wabnig E, Feistritzer C, Eder IE, Salvador C, Puhr M, Culig Z, Massoner P, Tiefenthaler M, Steurer M, Konwalinka G - PLoS ONE (2015)

Caspases are activated after AMF treatment.Capase -8, -9, and -3/-7 activity was measured using a luminometric assay after treatment with various concentrations of AMF for different time periods (4A). n = 3 (number of independent experiments carried out in triplicates). Furthermore Western blot analyses were performed for caspase activation and additionally for cleaved PARP, as a marker for apoptosis induction (4B). n = 3 (number of independent experiments, respresentative blots are shown)
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4334520&req=5

pone.0117806.g004: Caspases are activated after AMF treatment.Capase -8, -9, and -3/-7 activity was measured using a luminometric assay after treatment with various concentrations of AMF for different time periods (4A). n = 3 (number of independent experiments carried out in triplicates). Furthermore Western blot analyses were performed for caspase activation and additionally for cleaved PARP, as a marker for apoptosis induction (4B). n = 3 (number of independent experiments, respresentative blots are shown)
Mentions: In order to investigate whether the extrinsic or intrinsic apoptosis pathway was involved in AMF-induced apoptosis, we assessed caspase 8, -9 and -3/-7 activity using luminometric assays. As shown in Fig. 4A, AMF induced activity of all tested caspases in a concentration- and time-dependent manner. In particular, caspase 8 was induced already 8 hours after application of 3 μm of AMF in CEM C7H2, whereas in HL-60 higher concentrations and a longer treatment (10 μM for 24 hours) were necessary to yield a similar effect. Analogous results were found for caspase 9 activity. Finally, also caspase 3/-7 activity was induced in both cell lines, with CEM C7H2 being the more sensitive one. These results were corroborated on protein basis utilizing Western blotting for cleaved PARP, caspase -3, -8 and procaspase-9 (Fig. 4B). Unfortunately, we had to stick to an anti-procaspase-9 antibody (due to technical reasons), which when cleaved/activated should be decreased at protein level. However, as anticipated we found a profound decrease of procaspase-9 levels, suggestive for activation, which was found as stated above in experiments for Fig. 4A. Hence, we conclude that AMF induces apoptosis via both the extrinsic and the intrinsic pathway, which has been found to be the case also with various naturally occurring and synthetic molecules [14,15,16].

Bottom Line: We have previously shown that AMF exerts antiproliferative effects on a variety of solid tumor cell lines.This effect was found also in G0-arrested cells.Finally, in our animal experiments utilizing male nu/nu Balb/c mice we found a significant growth retardation, which was immunohistochemically associated with a significantly lower number of KI67-positive cells and caspase-3 induction in AMF-treated mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine IV, Medical University of Innsbruck, Innsbruck, Austria.

ABSTRACT
Akacid medical formulation (AMF) is an oligoguanidine that exerts biocidal activity against airborne and surface microorganisms including bacteria, viruses, fungi, and molds, while showing relatively low toxicity to humans. We have previously shown that AMF exerts antiproliferative effects on a variety of solid tumor cell lines. In this study we raised the question whether AMF could also substantially inhibit cell growth or induce apoptosis in cell lines derived from hematologic malignancies such as leukemia or lymphoma. We found that AMF has antiproliferative effects on various hematologic cell lines derived from human leukemia and lymphoma. Additionally, we show that AMF induces apoptosis in leukemia cell lines not only via the extrinsic and intrinsic pathway, but also in a caspase-independent manner. This effect was found also in G0-arrested cells. Finally, in our animal experiments utilizing male nu/nu Balb/c mice we found a significant growth retardation, which was immunohistochemically associated with a significantly lower number of KI67-positive cells and caspase-3 induction in AMF-treated mice.

No MeSH data available.


Related in: MedlinePlus