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In Vitro Investigations on the Toxicity and Cell Death Induced by Tamoxifen on Two Non-Breast Cancer Cell Types.

Majumdar SK, Valdellon JA, Brown KA - J. Biomed. Biotechnol. (2001)

Bottom Line: Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations.Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis).Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis.

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ABSTRACT
Tamoxifen, a potent anticancer agent known to interrupt the enhanced estrogen activity of malignant mammary gland cells, was recently approved by the Food and Drug Administration (FDA)for the treatment of breast cancer. In this investigation, the toxic effects of tamoxifen were evaluated through cell multiplication, and cytological, surface ultrastructural, and biochemical studies on human cervical carcinoma cells (HeLa)and/or murine erythroleukemic (MEL) cells (BB-88). Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations. The drug also triggered morphological and biochemical changes as revealed by light microscopy, scanning electron microscopy (SEM), fluorescence microscopy, Nucleosome ELISA, and the DNA smear pattern. Cytological observations showed nuclear condensation, cell shrinkage, multinucleation, and apoptotic bodies. Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis). Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis. Tamoxifen treated cells demonstrated internucleosomal damages of the genomic DNA and DNA fragmentations, evidenced by an increase in free nucleosomes, and distinctive DNA smear patterns on the agarose gel.

No MeSH data available.


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Agarose gel (0.8%) of DNA extracted from BB-88 cells: Lane 1, KB Ladder; Lane 2, Control (12 hours); Lane 3, tamoxifen treatment (12 hours, 10 μg/ml); Lane 4, tamoxifen treatment (12 hours,15 μg/ml). Only a faint DNA smearing is visible in the 15 μg/ml treatment group.
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Figure 7: Agarose gel (0.8%) of DNA extracted from BB-88 cells: Lane 1, KB Ladder; Lane 2, Control (12 hours); Lane 3, tamoxifen treatment (12 hours, 10 μg/ml); Lane 4, tamoxifen treatment (12 hours,15 μg/ml). Only a faint DNA smearing is visible in the 15 μg/ml treatment group.

Mentions: DNA was isolated from tamoxifen treated BB-88 cells and subjected to agarose gel electrophoresis to visualize the DNA laddering pattern. Tamoxifen induced degradation of genomic DNA was indicated by DNA smear patterns that are different from those of the controls. Tamoxifen treatment lanes revealed banding in the first half of the gel similar to the control, but visible DNA smearing patterns continued, contiguous to the smaller KB ladder fragments (see Figure 7). A faint DNA smearing observed in lane 4 (12 hours/ 15 μg/ml) was possibly due to the drastic effect of the drug on the cell viability which might have resulted in a severe degradation of nucleosomes.


In Vitro Investigations on the Toxicity and Cell Death Induced by Tamoxifen on Two Non-Breast Cancer Cell Types.

Majumdar SK, Valdellon JA, Brown KA - J. Biomed. Biotechnol. (2001)

Agarose gel (0.8%) of DNA extracted from BB-88 cells: Lane 1, KB Ladder; Lane 2, Control (12 hours); Lane 3, tamoxifen treatment (12 hours, 10 μg/ml); Lane 4, tamoxifen treatment (12 hours,15 μg/ml). Only a faint DNA smearing is visible in the 15 μg/ml treatment group.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC129061&req=5

Figure 7: Agarose gel (0.8%) of DNA extracted from BB-88 cells: Lane 1, KB Ladder; Lane 2, Control (12 hours); Lane 3, tamoxifen treatment (12 hours, 10 μg/ml); Lane 4, tamoxifen treatment (12 hours,15 μg/ml). Only a faint DNA smearing is visible in the 15 μg/ml treatment group.
Mentions: DNA was isolated from tamoxifen treated BB-88 cells and subjected to agarose gel electrophoresis to visualize the DNA laddering pattern. Tamoxifen induced degradation of genomic DNA was indicated by DNA smear patterns that are different from those of the controls. Tamoxifen treatment lanes revealed banding in the first half of the gel similar to the control, but visible DNA smearing patterns continued, contiguous to the smaller KB ladder fragments (see Figure 7). A faint DNA smearing observed in lane 4 (12 hours/ 15 μg/ml) was possibly due to the drastic effect of the drug on the cell viability which might have resulted in a severe degradation of nucleosomes.

Bottom Line: Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations.Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis).Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
Tamoxifen, a potent anticancer agent known to interrupt the enhanced estrogen activity of malignant mammary gland cells, was recently approved by the Food and Drug Administration (FDA)for the treatment of breast cancer. In this investigation, the toxic effects of tamoxifen were evaluated through cell multiplication, and cytological, surface ultrastructural, and biochemical studies on human cervical carcinoma cells (HeLa)and/or murine erythroleukemic (MEL) cells (BB-88). Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations. The drug also triggered morphological and biochemical changes as revealed by light microscopy, scanning electron microscopy (SEM), fluorescence microscopy, Nucleosome ELISA, and the DNA smear pattern. Cytological observations showed nuclear condensation, cell shrinkage, multinucleation, and apoptotic bodies. Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis). Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis. Tamoxifen treated cells demonstrated internucleosomal damages of the genomic DNA and DNA fragmentations, evidenced by an increase in free nucleosomes, and distinctive DNA smear patterns on the agarose gel.

No MeSH data available.


Related in: MedlinePlus